Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a real-time PCR-based in vitro diagnostic test with automated sample processing and result interpretation based on fluorescence detection. There is no mention of AI or ML in the device description, intended use, or performance studies. The interpretation appears to be based on predefined thresholds for fluorescence signals.

Therapeutic

No.
The device is an in vitro diagnostic test for the detection and identification of microbial nucleic acids from CSF specimens, intended as an aid in diagnosis and not for treatment or patient management decisions.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states, "The QIAstat-Dx Meningitis/Encephalitis (ME) Panel is a qualitative multiplexed nucleic acid real-time PCR based in vitro diagnostic test..." and, "The QIAstat-Dx ME Panel is indicated as an aid in the diagnosis of meningitis and/or encephalitis".

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic test that includes physical cartridges with reagents and is used with a dedicated hardware analyzer (QIAstat-Dx Analyzer 1.0). It is not solely software.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

The "Intended Use / Indications for Use" section explicitly states: "The QIAstat-Dx Meningitis/Encephalitis (ME) Panel is a qualitative multiplexed nucleic acid real-time PCR based in vitro diagnostic test intended for use with the QIAstat-Dx Analyzer 1.0."

This statement directly identifies the device as an in vitro diagnostic test.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The QIAstat-Dx® Meningitis/Encephalitis (ME) Panel is a qualitative multiplexed nucleic acid real-time PCR-based in vitro diagnostic test intended for use with the QIAstat-Dx Analyzer 1.0. The QIAstat-Dx ME Panel is capable of simultaneous detection and identification of multiple bacterial, viral, and yeast nucleic acids from cerebrospinal fluid (CSF) specimens obtained via lumbar puncture from individuals with signs and/or symptoms of meningitis and/or encephalitis.

The following organisms are identified using the QIAstat-Dx ME Panel: Enterovirus, Escherichia coli K1, Haemophilus influenzae, Listeria monocytogenes, Neisseria meningitidis (encapsulated), Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, and Cryptococcus neoformans/gattii*.

The QIAstat-Dx ME Panel is indicated as an aid in the diagnosis of specific agents of meningitis and/or encephalitis and results must be used in conjunction with other clinical, epidemiological, and laboratory data. Results from the QIAstat-Dx ME Panel are not intended to be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with organisms not included in the QIAstat-Dx ME Panel. The agent or agents detected may not be the definite cause of the disease. Negative results do not preclude central nervous system (CNS) infection.

Not all agents of CNS infection are detected by this test and sensitivity in clinical use may differ from that described in the instructions for use.

The QIAstat-Dx ME Panel is not intended for testing of specimens collected from indwelling CNS medical devices.

The QIAstat-Dx ME Panel is intended to be used in conjunction with standard of care culture for organism recovery, serotyping, and antimicrobial susceptibility testing.

Cryptococcus neoformans and Cryptococcus gattii are not differentiated.

Product codes

PLO

Device Description

The QIAstat-Dx® Meningitis/Encephalitis (ME) Panel is part of the QIAstat-Dx Meningitis/Encephalitis system and works with the QIAstat-Dx Analyzer 1.0.

The QIAstat-Dx ME Panel is intended to be used with cerebrospinal fluid (CSF) specimens.

Once the cartridge has been inserted into the instrument, the test starts automatically and runs for approximately 80 minutes. When the test is finished, the cartridge is removed by the user and discarded. The QIAstat-Dx Analyzer 1.0 automatically interprets test results and displays a summary on the analyzer display screen. The results can be printed using a connected printer, if needed. The detected analytes are displayed in red. For other analytes tested, they are displayed in green if not detected or in gray if not applicable or invalid. The analyzer will report if an error occurs during processing, in which case the test will fail and no results will be provided (screen will show "FAIL").

QIAstat-Dx consists of single-test cartridges with pre-packaged reagents including both wet and dry chemistry necessary to perform the sample preparation, nucleic acid amplification and detection to be used in conjunction with the QIAstat-Dx Analyzer 1.0. All sample preparation and assay steps are performed within the cartridge, so the user does not need to manipulate any reagent during the test. This eliminates exposure of the user or the Analyzer to chemicals contained in the cartridge during the test and up to the disposal of used cartridges.

Within the cartridge, multiple steps are automatically performed in sequence by using pneumatic pressure and a multiport valve to transfer the sample and fluids via the Transfer Chamber (TC) to their intended destinations. Following the introduction of the sample from a disposable transfer pipette, the following assay steps occur automatically and sequentially:

Resuspension of air-dried internal control and Proteinase K (ProtK) enzyme using provided buffer and mixing with the liquid sample (IC Cavity and ProtK Cavity);
Cell lysis using mechanical (rotation) and chemical (chaotropic and isotonic) means (lysis chamber):
Membrane-based nucleic acid purification from Lysate by:
- Mixing lysate with binding buffer and capturing on the membrane (purification chamber);
- First washing of membrane to remove bound proteins (purification chamber and waste chamber);
- Second washing of membrane to leave only bound nucleic acids (purification chamber and waste chamber);
- Rinsing of Transfer Chamber (TC) using the rinsing buffer before introduction of the eluate (Transfer Chamber);
Drying of membrane with bound nucleic acids with an air flow generated by a high flow vacuum pump (purification chamber); and
- Elution of nucleic acids with elution buffer (purification chamber and TC);
Mixing of the purified nucleic acid (eluate) with lyophilized "Master Mix" reagents (Dry chemistry container (DCC) and TC);
Sequential transfer of defined aliquots of mixed eluate/Master Mix from the Transfer Chamber to each of eight Reaction Chambers containing the specified, air-dried primers and probes;
Within each Reaction Chamber, real-time, multiplex PCR ("rtPCR") testing is performed. Increase in fluorescence (indicative of detection of each target analyte) is detected directly within each Reaction Chamber; and
The detected signal per fluorescent marker per Reaction Chamber is then used by the system software to generate the assay result.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Central nervous system (cerebrospinal fluid obtained via lumbar puncture)

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Prospective Specimens Testing:
A total of 1737 prospective residual CSF specimens were enrolled into the clinical study, of which 205 were withdrawn. The final dataset consisted of 1524 prospective specimens of which 552 (36.2%) were frozen before testing and 972 (63.8%) were tested fresh. Testing occurred between March 2022 and March 2023. Specimens were collected from patients with signs and symptoms of meningitis and/or encephalitis.
Residual CSF specimens were tested with the QIAstat-Dx ME Panel and two types of comparator methods (an FDA-cleared molecular comparator or two validated end point PCRs followed by bidirectional sequencing (BDS) for selected targets). All targets were compared to the FDA-cleared molecular method except Streptococcus pneumoniae and Streptococcus pyogenes which were compared against two validated end point PCRs followed by bidirectional sequencing. The standard of care (SoC) testing varied across all sites but included bacterial culture, Laboratory Developed PCR test (LDT), FDA-cleared molecular methods and Cryptococcus antigen screen and culture. Standard of care (SoC) culture results were collected to allow an assessment of clinical sensitivity and specificity and were investigated in cases of discordant result.

Archived Specimens Testing:
A total of 195 retrospective archived specimens were enrolled into the study to test analytes with low prevalence in the prospective arm. One hundred and fifty-four (154) archived specimens were excluded from the analysis as positivity was not confirmed by the comparator method. A total of 41 evaluable archived specimens were used in the analysis.

Contrived Specimens Testing:
Contrived specimens were used to support nine targets on the panel due to insufficient positive specimens from clinical collections. Contrived specimens were prepared by spiking five different quantified strains representative of the genetic diversity of each pathogen. For each pathogen, the LoD concentration was manufactured at 2x (at least 50%) and 5x LoD spiked into screened individual unique samples of negative CSF. Contrived specimens were tested alongside negative specimens in a blinded manner.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Non-clinical Studies:

Limit of Detection (LoD): The LoD for each QIAstat-Dx ME Panel target was assessed using 26 pathogen strains. LoD was confirmed at ≥95% detection rate.
Analytical Reactivity (Inclusivity): 130 strains covering 10 different analytical strains for each targeted organism or relevant species were tested. 125 out of 130 pathogen strains were successfully detected in vitro. In silico analysis demonstrated complete coverage for all main subtypes of target organisms.
Analytical Specificity (Exclusivity): On-Panel (13) and Off-Panel (115) strains were tested. All On-Panel pathogens showed specific detection, and all Off-Panel pathogens showed negative results, except for a few instances: Haemophilus haemolyticus showed cross-reactivity with Haemophilus influenzae (≥1.00E+03 CFU/mL), and Cryptococcus wingfieldii, Cryptococcus flavescens, and Cryptococcus amylolentus showed cross-reactivity with Cryptococcus neoformans/gattii (≥1.00E+01 CFU/mL, ≥4.00E+03 CFU/mL, ≥1.00E+01 CFU/mL resp.). In silico analysis pointed to 6 potential cross-reactions (Streptococcus pseudopneumoniae with Streptococcus pneumoniae; Listeria innocua with Listeria monocytogenes; Haemophilus haemolyticus with Haemophilus influenzae; Cryptococcus amylolentus, Cryptococcus depauperatus, and Cryptococcus wingfieldii with Cryptococcus neoformans/gattii).
Interfering Substances: Evaluated common endogenous and exogenous substances. Bleach caused inhibition at 1.0% v/v and 0.1% v/v, but no interference at 0.01% v/v. Other substances showed no interference.
Microbial Interference: Non-target organisms at challenging concentrations (10^5 units/mL for viral, 10^6 CFU/mL for bacterial) were tested with spiked target organisms at 3x LoD. All target organisms were successfully detected.
Competitive Inhibition: Clinically relevant co-infection testing demonstrated that when at least two QIAstat-Dx ME Panel pathogens of different concentrations are simultaneously present, all targets can be detected.
Carryover: No carryover between samples was observed using alternating high-positive and negative samples on two instruments.
Sample Stability: CSF specimens are stable at room temperature for up to 24 hours (15-25℃) and refrigerated for up to 7 days (2-8℃).
Matrix Equivalency: LoD in clinical CSF matrix was equivalent to artificial CSF for 18 out of 26 pathogen strains; 8 required new LoD determination.
Precision (Repeatability and Reproducibility): Multi-site study with 3 sites, 5 non-consecutive days, 6 replicates per day. Demonstrated successful precision with repeatable pathogen calling.

Clinical Performance:

Overall Prevalence: 1527 specimens, 4.3% positivity rate.
Prospective Clinical Performance:
- Sample size: 1524 prospective specimens (972 fresh, 552 frozen).
- Escherichia coli K1: Overall PPA 50.0% (2/4), NPA 100.0% (1520/1520).
- Haemophilus influenzae: Overall PPA 80.0% (4/5), NPA 99.8% (1516/1519).
- Listeria monocytogenes: Overall PPA 80.0% (4/5), NPA 100.0% (1519/1519).
- Neisseria meningitidis (encapsulated): Overall PPA 100.0% (1/1), NPA 99.9% (1522/1523).
- Streptococcus agalactiae: Overall PPA 100.0% (3/3), NPA 100.0% (1521/1521).
- Streptococcus pneumoniae: Overall PPA 100.0% (8/8), NPA 99.6% (1360/1365).
- Streptococcus pyogenes: Overall PPA N/A (0/0), NPA 100.0% (1291/1291).
- Enterovirus (EV): Overall PPA 91.7% (22/24), NPA 99.9% (1499/1500).
- Cryptococcus gattii / Cryptococcus neoformans (not differentiated): Overall PPA 57.1% (4/7), NPA 99.9% (1515/1517).
Archived Specimens Testing:
- Sample size: 41 evaluable archived specimens.
- Overall Panel: PPA 100.0% (41/41), NPA 100.0% (314/314).
- PPA for individual bacteria and fungi/yeast:
  - Escherichia coli Kl: 100.0% (2/2)
  - Haemophilus influenzae: 100.0% (6/6)
  - Listeria monocytogenes: N/A (0/0)
  - Neisseria meningitidis (encapsulated): 100.0% (3/3)
  - Streptococcus agalactiae: 100.0% (9/9)
  - Streptococcus pneumoniae: 100.0% (4/4)
  - Streptococcus pyogenes: N/A (0/0)
  - Enterovirus (EV): 100.0% (9/9)
  - Cryptococcus gattii / Cryptococcus neoformans (not differentiated): 100.0% (8/8)
Co-Infections: No specimens with multiple detections were identified by the QIAstat-Dx ME Panel.
Clinical Sensitivity and Specificity against Culture (for bacterial and fungal/yeast analytes): Ranges from 50.0% to 100.0% for sensitivity and 90.0% to 100.0% for specificity across various analytes and sample categories.
Validity of results: Initial failure rate for prospective fresh was 2.7%, prospective frozen 1.3%, and archived 1.7%. Final success rate after retest was 99.7% for prospective fresh, 99.6% for prospective frozen, and 100.0% for archived samples.
Contrived Specimens Testing: Achieved ≥95% positive results for all prepared contrived samples at 2xLoD and 5xLoD for all tested analytes.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Prospective Clinical Performance (PPA/NPA):

Escherichia coli K1: PPA 50.0% (95% CI: 15.0-85.0), NPA 100.0% (95% CI: 99.7-100.0)
Haemophilus influenzae: PPA 80.0% (95% CI: 37.6-96.4), NPA 99.8% (95% CI: 99.4-99.9)
Listeria monocytogenes: PPA 80.0% (95% CI: 37.6-96.4), NPA 100.0% (95% CI: 99.7-100.0)
Neisseria meningitidis (encapsulated): PPA 100.0% (95% CI: 20.7-100.0), NPA 99.9% (95% CI: 99.6-100.0)
Streptococcus agalactiae: PPA 100.0% (95% CI: 43.9-100.0), NPA 100.0% (95% CI: 99.7-100.0)
Streptococcus pneumoniae: PPA 100.0% (95% CI: 67.6-100.0), NPA 99.6% (95% CI: 99.1-99.8)
Streptococcus pyogenes: PPA N/A (0/0), NPA 100.0% (95% CI: 99.7-100.0)
Enterovirus (EV): PPA 91.7% (95% CI: 74.2-97.7), NPA 99.9% (95% CI: 99.6-100.0)
Cryptococcus gattii / Cryptococcus neoformans (not differentiated): PPA 57.1% (95% CI: 25.0-84.2), NPA 99.9% (95% CI: 99.5-100.0)

Archived Clinical Performance (PPA/NPA):

Overall Panel: PPA 100.0% (95% CI: 91.4-100.0), NPA 100.0% (95% CI: 98.8-100.0)

Clinical Sensitivity and Specificity against Culture (Selected Bacterial and Fungal/Yeast Analytes):

Escherichia coli Kl:
- Archived: Sensitivity 100.0% (95% CI: 20.7-100.0), Specificity 90.9% (95% CI: 62.3-98.4)
- Prospective Fresh: Sensitivity 50.0% (95% CI: 9.5-90.5), Specificity 100.0% (95% CI: 99.5-100.0)
Haemophilus influenzae:
- Archived: Sensitivity 100.0% (95% CI: 20.7-100.0), Specificity 90.9% (95% CI: 62.3-98.4)
- Prospective Frozen: Sensitivity 100.0% (95% CI: 43.9-100.0), Specificity 99.7% (95% CI: 98.4-99.9)
Listeria monocytogenes:
- Prospective Fresh: Sensitivity 100.0% (95% CI: 20.7-100.0), Specificity 100.0% (95% CI: 99.5-100.0)
- Prospective Frozen: Sensitivity 66.7% (95% CI: 20.8-93.9), Specificity 100.0% (95% CI: 98.9-100.0)
Neisseria meningitidis (encapsulated):
- Archived: Sensitivity 100.0% (95% CI: 34.2-100.0), Specificity 90.0% (95% CI: 59.6-98.2)
Streptococcus agalactiae:
- Prospective Fresh: Sensitivity 100.0% (95% CI: 20.7-100.0), Specificity 99.9% (95% CI: 99.3-100.0)
- Prospective Frozen: Sensitivity 100.0% (95% CI: 20.7-100.0), Specificity 100.0% (95% CI: 98.9-100.0)
Streptococcus pneumoniae:
- Prospective Frozen: Sensitivity 100.0% (95% CI: 43.9-100.0), Specificity 99.7% (95% CI: 98.4-99.9)
Cryptococcus gattii / Cryptococcus neoformans (not differentiated):
- Archived: Sensitivity 100.0% (95% CI: 34.2-100.0), Specificity 100.0% (95% CI: 20.7-100.0)
- Prospective: Sensitivity 100.0% (95% CI: 20.7-100.0), Specificity 98.5% (95% CI: 94.6-99.6)

Predicate Device(s)

K160462

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3970 Device to detect and identify microbial pathogen nucleic acids in cerebrospinal fluid.

(a)
Identification. A device to detect and identify microbial pathogen nucleic acids in cerebrospinal fluid is a qualitative in vitro device intended for the detection and identification of microbial-associated nucleic acid sequences from patients suspected of meningitis or encephalitis. A device to detect and identify microbial pathogen nucleic acids in cerebrospinal fluid is intended to aid in the diagnosis of meningitis or encephalitis when used in conjunction with clinical signs and symptoms and other clinical and laboratory findings.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include detailed device description documentation, including the device components, ancillary reagents required but not provided, and a detailed explanation of the methodology, including primer/probe sequence, design, and rationale for sequence selection.
(2) Premarket notification submissions must include detailed documentation from the following analytical studies: Analytical sensitivity (limit of detection), inclusivity, reproducibility, interference, cross reactivity, and specimen stability.
(3) Premarket notification submissions must include detailed documentation from a clinical study. The study, performed on a study population consistent with the intended use population, must compare the device performance to results obtained from well-accepted comparator methods.
(4) Premarket notification submissions must include detailed documentation for device software, including, but not limited to, software applications and hardware-based devices that incorporate software.
(5) The Intended Use statement in the device labeling must include a statement that the device is intended to be used in conjunction with standard of care culture.
(6) A detailed explanation of the interpretation of results and acceptance criteria must be included in the device's 21 CFR 809.10(b)(9) compliant labeling.
(7) The device labeling must include a limitation stating that the negative results do not preclude the possibility of central nervous system infection.
(8) The device labeling must include a limitation stating that device results are not intended to be used as the sole basis for diagnosis, treatment, or other patient management decisions.
(9) The device labeling must include a limitation stating that positive results do not mean that the organism detected is infectious or is the causative agent for clinical symptoms.
(10) As part of the risk management activities performed as part of your 21 CFR 820.30 design controls, you must document an appropriate end user device training program that will be offered as part of your efforts to mitigate the risk of failure to correctly operate the instrument.

Summary

0

October 29, 2024

Image /page/0/Picture/1 description: The image shows the logos of the Department of Health and Human Services and the Food and Drug Administration (FDA). The Department of Health and Human Services logo is on the left, and the FDA logo is on the right. The FDA logo includes the FDA acronym in a blue square, followed by the words "U.S. Food & Drug Administration" in blue text.

QIAGEN GmbH % Sonia Pablo Pablo Senior Manager, Regulatory Affairs STAT-Dx Life, S.L. (A QIAGEN Company) Carrer Baldiri Reixac. 4 Barcelona, 08028 Spain

Re: K242256

Trade/Device Name: QIAstat-Dx Meningitis/Encephalitis (ME) Panel Regulation Number: 21 CFR 866.3970 Regulation Name: Device To Detect And Identify Microbial Pathogen Nucleic Acids In Cerebrospinal Fluid Regulatory Class: Class II Product Code: PLO Dated: July 29, 2024 Received: July 31, 2024

Dear Sonia Pablo Pablo:

We have reviewed your section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (the Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database available at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device"

1

(https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30. Design controls; 21 CFR 820.90. Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review. the OS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rue"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-devices/device-advicecomprehensive-regulatory-assistance/unique-device-identification-system-udi-system.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

2

Sincerely, Bryan M. Grabias -S

Digitally signed by Bryan M. Grabias -S Date: 2024.10.29 15:52:56 -04'00'

Bryan Grabias Acting Branch Chief Bacterial Respiratory and Medical Countermeasures Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

3

Indications for Use

510(k) Number (if known) K242256

Device Name QIAstat-Dx Meningitis/Encephalitis (ME) Panel

Indications for Use (Describe)

The QIAstat-Dx Meningitis/Encephalitis (ME) Panel is a qualitative multiplexed nucleic acid real-time PCR based in vitro diagnostic test intended for use with the QIAstat-Dx Analyzer 1.0. The QIAstat-Dx ME Panel is capable of simultaneous detection and identification of multiple bacterial, viral, and yeast nucleic acids from cerebrospinal fluid (CSF) specimens obtained via lumbar puncture from individuals with signs and/or symptoms of meningitis and/or encephalitis.

The following organisms are identified using the OlAstat-Dx ME Panel: Enterovirus, Escherichia coli K1, Haemophilus influenzae, Listeria monocytogenes, Neisseria meningitidis (encapsulated), Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, and Cryptococcus neoformans/gattii*.

The QIAstat-Dx ME Panel is indicated as an aid in the diagnosis of meningitis and/or encephalitis and results must be used in conjunction with other clinical, endemiological, and laboratory data. Results from the OlAstat-Dx ME Panel are not intended to be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with organisms not included in the QIAstat-Dx ME Panel. The agents detected may not be the definite cause of the disease. Negative results do not preclude central nervous system infection.

Not all agents of central nervous system infection are detected by this test and sensitivity in clinical use may differ from that described in the instructions for use.

The QIAstat-Dx ME Panel is not intended for testing specimens collected from indwelling central nervous system medical devices.

The QIAstat-Dx ME Panel is intended to be used in conjunction with standard of care culture for organism recovery, serotyping, and antimicrobial susceptibility testing.

*Cryptococcus neoformans and Cryptococcus gattii are not differentiated.

Type of Use (Select one or both, as applicable)

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

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510(k) Summary

General Information

| Submitted by: | QIAGEN GmbH
QIAGEN Strasse 1
Hilden, Germany 40724 |
|-------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Contact Person: | Sonia Pablo
Senior Manager, Regulatory Affairs
STAT-Dx Life, S.L. (A QIAGEN Company)
Carrer Baldiri Reixac, 4
08028 Barcelona
Spain
Phone: +34 696 85 81 85
Email: sonia.pablo@qiagen.com |
| Date Prepared: | October 28, 2024 |
| Device Name: | QIAstat-Dx® Meningitis/Encephalitis (ME) Panel |
| Classification: | 21 CFR 866.3970 - Device to detect and identify microbial pathogen nucleic acids in cerebrospinal fluid. |
| Product Code: | PLO |
| Predicate Device: | |

Manufacturer	Product Name	De Novo/510(k) No.
BioFire Diagnostics, LLC	FilmArray Meningitis/Encephalitis (ME) Panel	K160462

5

Device Description

The QIAstat-Dx® Meningitis/Encephalitis (ME) Panel is part of the QIAstat-Dx Meningitis/Encephalitis system and works with the OIAstat-Dx Analyzer 1.0.

The QIAstat-Dx ME Panel is intended to be used with cerebrospinal fluid (CSF) specimens.

Once the cartridge has been inserted into the instrument, the test starts automatically and runs for approximately 80 minutes. When the test is finished, the cartridge is removed by the user and discarded. The OIAstat-Dx Analyzer 1.0 automatically interprets test results and displays a summary on the analyzer display screen. The results can be printed using a connected printer, if needed. The detected analytes are displayed in red. For other analytes tested, they are displayed in green if not detected or in gray if not applicable or invalid. The analyzer will report if an error occurs during processing, in which case the test will fail and no results will be provided (screen will show "FAIL").

QIAstat-Dx consists of single-test cartridges with pre-packaged reagents including both wet and dry chemistry necessary to perform the sample preparation, nucleic acid amplification and detection to be used in conjunction with the QIAstat-Dx Analyzer 1.0. All sample preparation and assay steps are performed within the cartridge, so the user does not need to manipulate any reagent during the test. This eliminates exposure of the user or the Analyzer to chemicals contained in the cartridge during the test and up to the disposal of used cartridges.

Within the cartridge, multiple steps are automatically performed in sequence by using pneumatic pressure and a multiport valve to transfer the sample and fluids via the Transfer Chamber (TC) to their intended destinations. Following the introduction of the sample from a disposable transfer pipette, the following assay steps occur automatically and sequentially:

Resuspension of air-dried internal control and Proteinase K (ProtK) enzyme using . provided buffer and mixing with the liquid sample (IC Cavity and ProtK Cavity);
Cell lysis using mechanical (rotation) and chemical (chaotropic and isotonic) ● means (lysis chamber):
Membrane-based nucleic acid purification from Lysate by: ●
- Mixing lysate with binding buffer and capturing on the membrane -(purification chamber);
- First washing of membrane to remove bound proteins (purification chamber and waste chamber);
- Second washing of membrane to leave only bound nucleic acids -(purification chamber and waste chamber);
- Rinsing of Transfer Chamber (TC) using the rinsing buffer before introduction of the eluate (Transfer Chamber);

6

Drying of membrane with bound nucleic acids with an air flow generated by a high flow vacuum pump (purification chamber); and
- Elution of nucleic acids with elution buffer (purification chamber and TC);
Mixing of the purified nucleic acid (eluate) with lyophilized "Master Mix" reagents ● (Dry chemistry container (DCC) and TC);
Sequential transfer of defined aliquots of mixed eluate/Master Mix from the ● Transfer Chamber to each of eight Reaction Chambers containing the specified, airdried primers and probes;
Within each Reaction Chamber, real-time, multiplex PCR ("rtPCR") testing is ● performed. Increase in fluorescence (indicative of detection of each target analyte) is detected directly within each Reaction Chamber; and
The detected signal per fluorescent marker per Reaction Chamber is then used by the system software to generate the assay result.

Intended Use

The QIAstat-Dx® Meningitis/Encephalitis (ME) Panel is a qualitative multiplexed nucleic acid real-time PCRbased in vitro diagnostic test intended for use with the QIAstat-Dx Analyzer 1.0. The QIAstat-Dx ME Panel is capable of simultaneous detection and identification of multiple bacterial, viral, and yeast nucleic acids from cerebrospinal fluid (CSF) specimens obtained via lumbar puncture from individuals with signs and/or symptoms of meningitis and/or encephalitis.

The following organisms are identified using the QIAstat-Dx ME Panel: Enterovirus, Escherichia coli K1, Haemophilus influenzae, Listeria monocvtogenes, Neisseria meningitidis (encapsulated), Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, and Cryptococcus neoformans/gattii*.

The OIAstat-Dx ME Panel is indicated as an aid in the diagnosis of specific agents of meningitis and/or encephalitis and results must be used in conjunction with other clinical, epidemiological, and laboratory data. Results from the QIAstat-Dx ME Panel are not intended to be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with organisms not included in the QIAstat-Dx ME Panel. The agent or agents detected may not be the definite cause of the disease. Negative results do not preclude central nervous system (CNS) infection.

Not all agents of CNS infection are detected by this test and sensitivity in clinical use may differ from that described in the instructions for use.

The QIAstat-Dx ME Panel is not intended for testing of specimens collected from indwelling CNS medical devices.

The QIAstat-Dx ME Panel is intended to be used in conjunction with standard of care culture for organism recovery, serotyping, and antimicrobial susceptibility testing.

7

Cryptococcus neoformans and Cryptococcus gattii are not differentiated.

Comparison of the QIAstat-Dx ME Panel and the Predicate Device

Similarities and differences between the QIAstat-Dx ME Panel and the predicate device are shown in Table 1.

Characteristic	Device	Predicate
Name	QIAstat-Dx ME Panel	FilmArray
Meningitis/Encephalitis (ME)
Panel
510(k) No.	K242256	K160462
Regulation	21 CFR 866.3970	21 CFR 866.3970
Product Code	PLO	PLO
Device Class	Class II	Class II
Similarities
Intended Use	The QIAstat-Dx®
Meningitis/Encephalitis (ME)
Panel is a qualitative multiplexed
nucleic acid real-time PCR-based
in vitro diagnostic test intended
for use with the QIAstat-Dx
Analyzer 1.0. The QIAstat-Dx
ME Panel is capable of
simultaneous detection and
identification of multiple
bacterial, viral, and yeast nucleic
acids from cerebrospinal fluid
(CSF) specimens obtained via
lumbar puncture from individuals
with signs and/or symptoms of
meningitis and/or encephalitis.

The following organisms are
identified using the QIAstat-Dx
ME Panel:

Bacteria:
• Escherichia coli K1
• Haemophilus influenzae
• Listeria monocytogenes | The FilmArray
Meningitis/Encephalitis (ME)
Panel is a qualitative
multiplexed nucleic acid-based
in vitro diagnostic test intended
for use with FilmArray,
FilmArray 2.0, and FilmArray
Torch systems. The FilmArray
ME Panel is capable of
simultaneous detection and
identification of multiple
bacterial, viral, and yeast
nucleic acids directly from
cerebrospinal fluid (CSF)
specimens obtained via lumbar
puncture from individuals with
signs and/or symptoms of
meningitis and/or encephalitis.

The following organisms are
identified using the FilmArray
ME Panel:

Bacteria:
• Escherichia coli K1
• Haemophilus influenzae
• Listeria monocytogenes |

	Table 1: Comparison of the QIAstat-Dx ME Panel with the predicate device
--	--------------------------------------------------------------------------	--	--	--	--	--

8

510(k) Premarket Notification

Page 5 of 35

Characteristic	Device	Predicate
	• Neisseria meningitidis
(encapsulated)	• Neisseria meningitidis
(encapsulated)
	• Streptococcus agalactiae	• Streptococcus agalactiae
	• Streptococcus pneumoniae	• Streptococcus pneumoniae
	• Streptococcus pyogenes
		Viruses:
	Virus:	• Enterovirus
	• Enterovirus	• Cytomegalovirus
		• Herpes simplex virus 1
	Yeast:	• Herpes simplex virus 2
	• Cryptococcus
neoformans/gattii*	• Human herpesvirus 6
		• Human parechovirus
		• Varicella zoster virus
	The QIAstat-Dx ME Panel is
indicated as an aid in the
diagnosis of specific agents of
meningitis and/or encephalitis and
results must be used in
conjunction with other clinical,
epidemiological, and laboratory
data. Results from the QIAstat-
Dx ME Panel are not intended to
be used as the sole basis for
diagnosis, treatment, or other
patient management decisions.
Positive results do not rule out co-
infection with organisms not
included in the QIAstat-Dx ME
Panel. The agent or agents
detected may not be the definite
cause of the disease. Negative
results do not preclude central
nervous system (CNS) infection.

Not all agents of CNS infection
are detected by this test and
sensitivity in clinical use may | | |
| | | Yeast: | |
| | | • Cryptococcus
neoformans/gattii | |
| | | The FilmArray ME Panel is
indicated as an aid in the
diagnosis of specific agents of
meningitis and/or encephalitis
and
results are meant to be used in
conjunction with other clinical,
epidemiological, and laboratory
data. Results from the
FilmArray ME Panel are not
intended to be used as the sole
basis for diagnosis, treatment, or
other patient management
decisions. Positive results do not
rule out co-infection with
organisms not included in the
FilmArray ME Panel. The agent
detected may not be the definite
cause of the disease. Negative
results do not preclude central
nervous system (CNS)
infection. Not all agents of CNS
infection are detected by this
test and sensitivity in clinical
use may differ from that | |
| Characteristic | Device | Predicate | |
| | differ from that described in the
instructions for use. | described in the package insert. | |
| | The QIAstat-Dx ME Panel is not
intended for testing of specimens
collected from indwelling CNS
medical devices. | The FilmArray ME Panel is not
intended for testing of
specimens collected from
indwelling CNS medical
devices. | |
| | The QIAstat-Dx ME Panel is
intended to be used in conjunction
with standard of care culture for
organism recovery, serotyping, and
antimicrobial susceptibility testing. | The FilmArray ME Panel is
intended to be used in
conjunction with standard of
care culture for organism
recovery,
serotyping, and antimicrobial
susceptibility testing. | |
| | * Cryptococcus neoformans and
Cryptococcus gattii are not
differentiated. | | |
| Specimen Type | Cerebrospinal Fluid | Cerebrospinal Fluid | |
| Analyte Detected | RNA/DNA | RNA/DNA | |
| Organisms
Detected | See above | See above | |
| Amplification and
Detection
Technology | PCR | PCR | |
| Differences | | | |
| Assay Controls | One internal control in each
cartridge to control for sample
processing that is subjected to all
nucleic acid extraction and
amplification steps similar to
patient samples. Labeling will
recommend use of negative and
positive external controls
regularly. Use transport media as
the external negative control, and
previously characterized positive
samples or negative sample
spiked with well characterized
target organisms as external
positive controls. | Two controls are included in
each reagent pouch to control
for sample processing and
both stages of PCR and melt
analysis. Labeling recommends
the use of external positive and
negative controls regularly.
Enteric transport media can be
used as an external negative
control, and previously
characterized positive samples
or negative samples spiked with
well characterized organisms as
external positive controls. | |
| Characteristic | Device | Predicate | |
| Assay Targets | The QIAstat-Dx ME Panel detects:

Bacteria:
• Streptococcus pyogenes | The FilmArray ME Panel detects:

Viruses:
• Cytomegalovirus
• Herpes simplex virus 1
• Herpes simplex virus 2
• Human herpesvirus 6
• Human parechovirus
• Varicella zoster virus | |
| | Nucleic Acid
Extraction
Technology | Extraction of nucleic acids using silica membrane | Extraction of nucleic acids using standard magnetic beads |
| | | QIAstat-Dx detection of amplified targets uses an increase in fluorescence to generate the assay results. | The FilmArray ME Panel uses melting curve analysis to confirm the identity of amplified targets to generate assay results. |
| | Operational | The sample is loaded straight into the cartridge. | The FilmArray ME Panel must rehydrate and prime the cartridge before use. |
| | Amplification and
Detection
Instrument
System | QIAstat-Dx Analyzer 1.0 | FilmArray Instruments |

9

10

Performance Characteristics - Non-clinical Studies

The studies presented have been performed to demonstrate the non-clinical performance of the QIAstat-Dx ME Panel.

Limit of Detection

The Limit of Detection (LoD) is defined as the lowest concentration at which >95% of samples tested generate a positive call.

The LoD for each QIAstat-Dx ME Panel target was assessed, using 26 pathogen strains, by analyzing dilutions of analytical samples prepared from stocks obtained from commercial suppliers. The LoD was determined using selected strains representing individual pathogens that are possible to detect with the QIAstat- Dx ME Panel. All sample dilutions were prepared using artificial CSF. To confirm the established LoD concentration, the required detection rate of all replicates was ≥95%. Additional testing of samples prepared using negative clinical CSF was conducted to assess equivalency.

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Individual LoD concentrations for each QIAstat-Dx ME Panel target are shown in Table 2.

| Pathogen | Strain | Source /
Catalog ID | LoD
Concentration* | Detection
Rate |
|------------------------------------------|--------------------------------------------------|--------------------------|-----------------------|-------------------|
| Escherichia coli Kl | Strain C5 [Bort];
018ac:K1:H7 | ATCC
700973 | 3.48E+02 CFU/mL | 30/30 |
| Escherichia coli K1 | NCTC 9001.
Serovar 01:K1:H7 | ATCC
11775 | 7.86E+02 CFU/mL | 30/30 |
| Haemophilus influenzae | type b (cap) | ATCC
10211 | 3.16E+02 CFU/mL | 32/32 |
| Haemophilus influenzae | Type e [strain AMC
36-A-7 | ATCC
8142 | 2.54E+03 CFU/mL | 30/30 |
| Listeria monocytogenes | Type 1/2b | ZeptoMetrix
0801534 | 1.86E+03 CFU/mL | 21/21 |
| Listeria monocytogenes | Type 4b. Strain Li 2 | ATCC
19115 | 6.64E+03 CFU/mL | 30/30 |
| Neisseria meningitidis
(encapsulated) | Serotype B. M2092 | ATCC
13090 | 8.28E-02 CFU/mL | 31/32 |
| Neisseria meningitidis
(encapsulated) | Serotype Y. M-112
[BO-6] | ATCC
35561 | 1.33E+01 CFU/mL | 30/30 |
| Streptococcus agalactiae | Z019 | ZeptoMetrix
0801545 | 1.75E+03 CFU/mL | 30/30 |
| Streptococcus agalactiae | G19 group B | ATCC
13813 | 3.38E+03 CFU/mL | 31/31 |
| Streptococcus pneumoniae | 19F | ZeptoMetrix
0801439 | 7.14E+02 CFU/mL | 29/30 |
| Streptococcus pneumoniae | Serotype 1. NCTC
7465 | ATCC
33400 | 6.22E-01 CFU/mL | 29/29 |
| Streptococcus pyogenes | Z472; Serotype M1 | ZeptoMetrix
0804351 | 1.80E+03 CFU/mL | 30/30 |
| Streptococcus pyogenes | Bruno [CIP
104226] | ATCC
19615 | 9.10E+01CFU/mL | 30/30 |
| Enterovirus A | Coxsackievirus
A16 | ZeptoMetrix
0810107CF | 3.79E+00 TCID50/mL | 31/31 |
| Enterovirus A | A6, species A.
Strain Gdula | ATCC
VR-1801 | 1.60E+02 TCID50/mL | 30/30 |
| Enterovirus B | Coxsackievirus B5 | ZeptoMetrix
0810019CF | 8.91E+01 TCID50/mL | 30/30 |
| Enterovirus B | Coxsackievirus A9,
species B | ZeptoMetrix
0810017CF | 4.36E+01 TCID50/mL | 28/29 |
| Enterovirus C | Coxsackievirus
A17, species C.
Strain G-12 | ATCC
VR-1023 | 1.58E+01 TCID50/mL | 30/30 |
| Enterovirus C | Coxsackievirus
A24. Starin DN-19 | ATCC
VR-583 | 4.99E+00 TCID50/mL | 30/30 |
| Pathogen | Strain | Source /
Catalog ID | LoD
Concentration* | Detection
Rate |
| Enterovirus D | EV 70, species D,
strain J670/71 | ATCC
VR-836 | $4.99E+01$ TCID50/mL | 30/31 |
| Enterovirus D | Enterovirus D68.
Strain US/MO/14-
18947 | ATCC
VR-1823 | $5.06E+02$ TCID50/mL | 30/30 |
| Cryptococcus neoformans | Serotype D strain
WM629, type
VNIV | ATCC
MYA-4567 | $2.21E+03$ CFU/mL | 31/31 |
| Cryptococcus neoformans | C. neoformans H99 | ATCC
208821 | $1.64E+02$ CFU/mL | 31/31 |
| Cryptococcus gattii | Serotype B strain
R272, type VGIIb | ATCC
MYA-4094 | $1.32E+04$ CFU/mL | 30/30 |
| Cryptococcus gattii | A6MR38 [CBS
11545] | ATCC
MYA-4877 | $2.60E+03$ CFU/mL | 29/29 |

Table 2: LoD concentrations for the strains tested with QIAstat-Dx ME Panel

12

*Highest LoD concentration from LoD/Matrix Equivalency studies is reported.

Analytical Reactivity (Inclusivity)

The Inclusivity (analytical reactivity) study extended the list of pathogen strains tested during the QIAstat-Dx ME Panel Limit of Detection (LoD) study to confirm the reactivity of the detection system in the presence of different strains of the same organisms at a concentration near or above the respective Limit of Detection.

A variety of clinically relevant strains of each target organism of the OIAstat-Dx ME Panel representing organism sub-types, strains, serotypes, and genotypes of different temporal and geographic diversity of each analyte were included in the study. Analytical Reactivity was performed in two steps (in vitro and in silico).

Based on in vitro (wet) and in silico analysis, the QIAstat-Dx ME Panel has demonstrated a broad coverage for all targets detected by the panel.

In vitro analysis

A total of 130 strains covering 10 different analytical strains for each targeted organism or relevant species were tested spiked in combined samples. Strains tested for inclusivity are shown in Table 3.

125 out of 130 pathogen strains were successfully detected by the QIAstat-Dx ME Panel when tested in vitro. Five strains were not detected by the assay (detailed in Table 4).

| Pathogen | Source
(Catalog ID) | Strain/Serotype | Lowest Concentration
Tested |
|------------------------------------------------------|------------------------------|-----------------------------------------------------------------|--------------------------------|
| Escherichia
coli K1 | ATCC (700973)a | Strain C5 [Bort]; O18ac:K1:H7 | 1x LoD |
| | ATCC (11775)a | NCTC 9001. Serovar O1:K1:H7 | 1x LoD |
| | NCTC (9007) | Strain Bi 7509/41; O7:K1:H- | 0.03x LoD |
| Pathogen | Source
(Catalog ID) | Strain/Serotype | Lowest Concentration
Tested |
| Haemophilus
influenzae | ATCC (23509) | NCDC Bi 7509-41 Serotype 07
:K1(L) :NM | 1:100 from stock |
| | ATCC (23511) | NCDC F 11119-41 | 0.03x LoD |
| | BEI Resources (NR-
17666) | O-2, U9-41 | 1:100 from stock |
| | BEI Resources (NR-
17674) | O-16, F1119-41 | 1:100 from stock |
| | ZeptoMetrix (0801905) | Z136 CTX-M-15 | 3x LoD |
| | NCTC (11101) | Sc15 02:K1:H6 | 3x LoD |
| | NCTC (9045) | Strain H61; O45:K1:H10 | 3x LoD |
| | ATCC (10211) a | type b (cap) | 1x LoD |
| | ATCC (8142) a | Type e [strain AMC 36-A-7] | 1x LoD |
| | ATCC (51907) | Non-typeable [strain Rd [KW20] | 0.014x LoD |
| | ATCC (11116) | Non-typeable [strain 180-a] | 3x LoD |
| | ATCC (9006) | Type a [strain AMC 36-A-3] | 3x LoD |
| | ATCC (31512) | Type b [strain Rab] | 0.016x LoD |
| | ATCC (49699) | Type c [strain C 9007] | 0.016x LoD |
| | ATCC (9008) | Type d [strain AMC 36-A-6] | 0.014x LoD |
| | ATCC (700223) | Type f [strain GA-1264] | 0.016x LoD |
| | ATCC (49766) | L-378 | 3x LoD |
| Listeria
monocytogenes | ZeptoMetrix (0801534) a | Type 1/2b | 1x LoD |
| | ATCC (19115) a | Type 4b. Strain Li 2 | 1x LoD |
| | ATCC (BAA-2659) | Type 1/2a. Strain 2011L-2676 | 3x LoD |
| | ATCC (19111) | Type 1/2a. Strain Li 20 | 3x LoD |
| | ZeptoMetrix (0804339) | Type 4b | 0.03x LoD |
| | ATCC (13932) | serotype 4b. Strain 1071/53 [LMG
21264, NCTC 10527] | 0.011x LoD |
| | ATCC (19114) | Li 23. Serotype 4a | 0.02x LoD |
| | BEI Resources (NR-
13237) | FSL J2-064 | 1:100 from stock |
| | ATCC (7644) | Gibson | 2x LoD |
| | ATCC (BAA-679) | EGDe | 0.026x LoD |
| Neisseria
meningitidis
(encapsulated) | ATCC (13090) a | Serotype B. M2092 [CIP 104218,
L. Cunningham] | 1x LoD |
| | ATCC (35561) a | Serotype Y. M-112 [BO-6] | 1x LoD |
| | ATCC (13077) | Serogroup A, M1027
[NCTC10025] | 0.03x LoD |
| | ATCC (13102) | Serogroup C, M1628 | 0.03x LoD |
| | ATCC (13113) | Serotype D. M158 [37A] | 3x LoD |
| | IDT (gBlock 77859371)b | sequence with variant ctrA gene | 1:1000 from stock |
| | ATCC (43744) | W135 | 0.03x LoD |
| | ATCC (BAA-335) | MC58 | 0.03x LoD |
| Streptococcus
agalactiae | ATCC (23255) | 79 Eur. Serogroup B | 1:1000 from stock |
| | ATCC (13092) | Serotype B. M997 [S-3250-L] | 1:1000 from stock |
| | ZeptoMetrix (0801545) a | Z019 | 1x LoD |
| | ATCC (13813) a | G19 group B | 1x LoD |
| | ATCC (12403) | Serotype III. Typing strain
D136C(3) [3 Cole 106, CIP 82.45] | 1.2x LoD |
| | ATCC (BAA-611) | 2603 V/R. Serotype V | 0.013x LoD |
| Pathogen | Source
(Catalog ID) | Strain/Serotype | Lowest Concentration
Tested |
| | ATCC (31475) | type III-ST283 | 3x LoD |
| | BEI Resources (NR-
43898) | MNZ929 | 1:100 from stock |
| | ATCC (12401) | Typing strain H36B - type Ib | 1.4x LoD |
| | ATCC (27591) | CDC SS700 [A909; 5541], type 1c | 0.014x LoD |
| | ATCC (49446) | 3139 [CNCTC 1/82] Serotype IV | 0.0127x LoD |
| | ZeptoMetrix (0801556) | Z023 | 3x LoD |
| | ZeptoMetrix (0801439) a | 19F | 1x LoD |
| | ATCC (33400) a | Serotype 1. NCTC 7465 | 1x LoD |
| Streptococcus
pneumoniae | ATCC (BAA-334) | Serotype 4. TIGR4 [JNR.7/87] | 0.03x LoD |
| | ATCC (BAA-341) | Serotype 5. SPN1439-106
[Colombia 5-19] | 0.03x LoD |
| | ATCC (10343) | Serotype 11A. Type 43 | 3x LoD |
| | ATCC (700672) | Serotype 14. VH14 | 0.03x LoD |
| | ATCC (700673) | Serotype 19A. Hungary 19A-6
[HUN663] | 0.026x LoD |
| | ZeptoMetrix (0804016) | Z319; 12F | 3x LoD |
| | ATCC (6303) | Diplococcus pneumoniae; Type 3.
Strain [CIP 104225] | 3x LoD |
| | ATCC (BAA-661) | DCC1476 [Sweden 15A-25] | 0.03x LoD |
| | ZeptoMetrix (0804351) a | Z472; Serotype M1 | 1x LoD |
| | ATCC (19615) a | Bruno [CIP 104226] | 1x LoD |
| | ZeptoMetrix (0801512) | Z018; Serotype M58 | 3x LoD |
| Streptococcus
pyogenes | ATCC (BAA-947) | Serotype M1. MGAS 5005 | 0.03x LoD |
| | ATCC (14289) | Lancefield's group A / C203 S | 3x LoD |
| | ATCC (12203) | NCTC 8709 (Type 6 glossy) | 0.03x LoD |
| | ATCC (12353) | Group a, type 12. Typing strain
T12 [F. Griffith SF 42] | 1:1000 from stock |
| | ATCC (12972) | Group a, type 14 | 0.03x LoD |
| | ATCC (8133) | Group a, type 23 | 3x LoD |
| | ATCC (12384) | C203 -Type 3 | 0.029x LoD |
| | ZeptoMetrix (0810107CF) | Coxsackievirus A16 | 1x LoD |
| Enterovirus A | ATCC (VR-1801) a | A6, species A. Strain Gdula | 1x LoD |
| | ATCC (VR-168) | A10. Μ.Κ. (Kowalik) | 3x LoD |
| | ATCC (VR-1432) | Enterovirus 71. Strain H | 3x LoD |
| | ZeptoMetrix (0810236CF) | Species A, Serotype EV-A71
(2003 Isolate) | 1:100 from stock |
| | BEI Resources (NR-471) | Tainan/4643/1998 | 3x LoD |
| | ATCC (VR-1550) | A2 F1 [Fleetwood] | 3x LoD ° |
| | ATCC (VR-673) | A7 - 275/58 | 3x LoD |
| | ATCC (VR-170) | A12 – Texas 12 | 1:100 from stock |
| | ATCC (VR-1775) | EV-A71. Strain BrCr | 3x LoD |
| Enterovirus B | ZeptoMetrix (0810019CF) | Coxsackievirus B5 | 1x LoD |
| | ZeptoMetrix (0810017CF) a | Coxsackievirus A9, species B | 1x LoD |
| | ATCC (VR-28) | Species B, Serotype CV-B1, Strain
Conn-5 | 3x LoD |
| | ATCC (VR-29) | Species B, Serotype CV-B2. Strain
Ohio-1 | 3x LoD |
| | ZeptoMetrix (0810075CF) | Coxsackievirus B4 | 1:100 from stock |
| | ZeptoMetrix (0810076CF) | Echovirus 6 | 1:100 from stock |
| | ZeptoMetrix (0810077CF) | Echovirus 9 | 1:100 from stock |
| | ZeptoMetrix (0810074CF) | Coxsackievirus B3 | 1:10000 from stock |
| Pathogen | Source
(Catalog ID) | Strain/Serotype | Lowest Concentration
Tested |
| | NCPV (0901047v) | Echovirus 18 | 3x LoD |
| | ATCC (VR-41) | Species B, Serotype E-11 | 3x LoD |
| Enterovirus C | ATCC (VR-1023)a | Coxsackievirus A17, species C.
Strain G-12 | 1x LoD |
| | ATCC (VR-583)a | Coxsackievirus A24. Starin DN-19 | 1x LoD |
| | ATCC (VR-850) | Coxsackievirus A21. Strain
Kuykendall [V-024-001-012] | 3x LoDc |
| | ATCC (VR-169) | A11 - Belgium-1 | 3x LoD |
| | ATCC (VR-1488) | A13 - Flores | 3x LoD |
| | ATCC (VR-182) | A22 - Chulman | 1:100 from stock |
| | ATCC (VR-178) | A20 - IH Pool 35 | 1:100 from stock |
| | ATCC (VR-176) | A18 - G-13 | 1:100 from stock |
| | NCTC (0812075v) | CV-A21. Strain H06452 472 | 3x LoD |
| | NCTC (0812074v) | CV-A21. Strain H06418 508 | 0.03x LoD |
| | ATCC (VR-836)a | EV 70, species D, strain J670/71 | 1x LoD |
| | ATCC (VR-1823)a | Enterovirus D68. Strain
US/MO/14-18947 | 1x LoD |
| | ZeptoMetrix (0810237CF) | Enterovirus 68. 2007 Isolate | 0.03x LoD |
| | ATCC (VR-1824) | Enterovirus D68. Strain US/IL/14-
18952 | 3x LoD |
| | ATCC (VR-1197) | D68. Strain F02-3607 Corn | 3x LoD |
| Enterovirus D | ZeptoMetrix (0810302CF) | Type 68 Major Group (09/2014
Isolate 2) | 1:100 from stock |
| | ATCC (VR-1825) | Enterovirus D68. Strain
US/KY/14-18953 | 3x LoD |
| | ATCC (VR-1826) | Enterovirus D68. Strain Fermon | 3x LoD |
| | BEI Resources (NR-
49130) | Enterovirus D68. US/MO/14-
18949 | 3x LoD |
| | BEI Resources (NR-
51998) | Enterovirus D68. USA/2018-23089 | 3x LoD |
| | ATCC (MYA-4567)a | Serotype D strain WM629, type
VNIV | 1x LoD |
| | ATCC (208821)a | H99 | 1x LoD |
| | ATCC (32045) | type strain, CBS 132 | 3x LoD |
| | ATCC (MYA-4564) | Serotype A strain WM148, type
VNI | 1:1000 from stock |
| | ATCC (13690) | M2092 | 1:100 from stock |
| Cryptococcus
neoformans | ATCC (MYA-4566) | Serotype AD strain WM628, type
VNIII | 1:1000 from stock |
| | ZeptoMetrix (0801803) | Serotype A | 3x LoD |
| | BEI Resources (NR-
50335) | NIH9hi90 | 1:100 from stock |
| | BEI Resources (NR-
50332) | NIH306 | 1:100 from stock |
| | BEI Resources (NR-
48776) | Var grubiiYL99α | 1:100 from stock |
| | ATCC (MYA-4094)a | Serotype B strain R272, type
VGIIb | 1x LoD |
| | ATCC (MYA-4877)a | A6MR38 | 1x LoD |
| Cryptococcus
gattii | ATCC (MYA-4560) | Serotype B strain WM179, type
VGI | 1:100 from stock |
| | ATCC (MYA-4562) | Serotype B strain WM161, type
VGIII | 1:1000 from stock |
| | ATCC (MYA-4563) | Serotype C strain WM779, type
VGIV | 1:1000 from stock |
| | ATCC (MYA-4138) | A1M R265 | 3x LoD |
| Pathogen | Source
(Catalog ID) | Strain/Serotype | Lowest Concentration
Tested |
| | ATCC (14248) | 110 [CBS 883] | 1:1000 from stock |
| | BEI Resources (NR-50184) | AIR265 | 1:100 from stock |
| | BEI Resources (NR-50195) | Alg166 | 1:100 from stock |
| | BEI Resources (NR-50198) | Alg254 | 1:100 from stock |

Table 3: Strains tested for Inclusivity

13

14

15

16

a Strains tested and evaluated during the Limit of Detection studies.

b Commercial artificial dsDNA fragment (gBlock) including ctrA gene sequence was tested due to unavailability of analytical sample for this variant (GenBank Accession ID HO156899).

6 Higher concentration tested to meet 100% detection rate (30xLoD for ATCC (VR-1550, VR-850 and VR-41) and 3xLoD for ZeptoMetrix (0804339)).

Pathogen	Strain/Serotype
Escherichia coli K1	NCDC Bi 7509-41 Serotype 07:K1(L):NM
Escherichia coli K1	Z136 CTX-M-15
Enterovirus C	CV-A21. Strain H06452 472
Enterovirus C	CV-A21. Strain H06418 508
Streptococcus agalactiae	Serotype III. Typing strain D136C(3) [3 Cole 106, CIP 82.45]

Table 4: Inclusivity Strains not detected by the QIAstat-Dx ME Panel

In silico analysis

An in silico prediction of the reactivity of all primers-probe oligonucleotide sequences included in the panel against sequences publicly available in databases was performed. The analysis demonstrated a complete coverage for the detection of all main subtypes (species and serotypes) for every On-Panel target organism included in the QIAstat-Dx ME Panel (Table 5).

Pathogens	Clinically relevant strains/subtypes detected
Neisseria meningitidis
(encapsulated)	Encapsulated serotypes (A, B, C, D, E, H, I, K, L, NG, W, W135, X,
Y, Z, 29E).
Cryptococcus gattii/
neoformans	Serotype A ( C. neoformans var neoformans ), serotype D ( C.
neoformans var grubii ), serotpyes B and C ( C. gattii including all
VGI,VGII, VGIII, VGIV molecular types).
Listeria monocytogenes	Serotypes 1/2a, 1/2b, 1/2c, 3a, 3b, 3c, 4a, 4b, 4c, 4d, 4e, 7.
Haemophilus influenzae	All encapsulated serotypes (a, b, c, d, e, f) and unencapsulated strains
(non-typeable, NTHi) including var. H. Aegyptus
Enterovirus	Coxsackievirus A (CV-A1 through CV-A24), coxsackievirus B (CV-
B1 through CV-B6), Echovirus (E-1 through E-33), Enterovirus A
(EV-A71, EV-A76, EV-A89 through EV-A92, EV-A119, EV-A120),
Enterovirus B (EV-B69, EV-B73 through EV-B75, EV-B79, EV-B80
through EV-B88, EV-B93, EV-B97, EV-B98, EV-B100, EV-B101,
EV-B106, EV-B107, EV-B111), Enterovirus C (EV-C96, EV-C99,
EV-C102, EV-C104, EV-C105, EV-C109, EV-C116 through EV-
C118), Enterovirus D (EV-D68, EV-D70, EV-D94), Poliovirus (PV-1
through PV-3).
Escherichia coli K1	K1 strains

Table 5: Organisms with predicted reactivity based on in silico analysis

17

Pathogens	Clinically relevant strains/subtypes detected
Rest of On-Panel organism
with no biological
subclassification (S.
pneumoniae, S. agalactiae, S.
pyogenes)	All genomic sequences available in databases detected

Analytical Specificity (Exclusivity)

The analytical specificity study was carried out by in vitro testing and in silico analysis to assess the potential cross-reactivity and exclusivity of the QIAstat-Dx ME Panel. On-Panel organisms were tested to assess the potential for intra-panel cross-reactivity and Off-Panel organisms were tested to evaluate cross-reactivity with organisms not covered by the panel content (panel exclusivity). The Off-Panel organisms were selected because they are clinically relevant (colonize the central nervous system or cause meningitis and/or encephalitis symptoms), are common skin flora or laboratory contaminants, are genetically similar to On-Panel analytes, or are microorganisms for which much of the population may have been infected.

After testing several On-Panel (13) and Off-Panel (115) strains, the QIAstat-Dx ME Panel demonstrated high specificity levels for all the rtPCR assays included in the device. Additional bioinformatic prediction analysis supported the specificity level, with few exceptions.

In vitro analysis

Samples were prepared by spiking potential cross-reactive organisms into artificial CSF matrix at 105 TCIDso/ml for viral targets, 105 CFU/mL for fungi / yeast target, and 100 CFU/mL for bacterial and fungi / yeast target, or the highest concentration possible based on the organism stock. The On-Panel and Off-Panel organisms tested are shown in Table 6.

All On-Panel pathogens resulted in specific detection, and all Off-Panel pathogens tested showed a negative result and no cross-reactivity was observed in the QIAstat-Dx ME Panel, except for the pathogens shown in Table 7.

Type	Pathogen	Strain/Serotype
On-Panel
Bacteria	Escherichia coli K1	Strain C5 [Bort];
O18ac:K1:H7
Bacteria	Haemophilus influenzae	Type e [strain AMC 36-A-
7]
Bacteria	Listeria monocytogenes	Type 4b. Strain Li 2

		Table 6: Off-Panel and On-Panel Analytical strains tested during Exclusivity

18

Type	Pathogen	Strain/Serotype
On-Panel
Bacteria	Neisseria meningitidis	Serotype Y. M-112 [BO-6]
Bacteria	Streptococcus pneumoniae	19F
Bacteria	Streptococcus agalactiae	Z019
Bacteria	Streptococcus pyogenes	Z472; Serotype M1
Virus	Enterovirus A1	A6, species A.
Strain Gdula
Virus	Enterovirus B1	Coxsackievirus B5
Virus	Enterovirus C1	Coxsackievirus A17,
species C. Strain G-12
Virus	Enterovirus D1	Enterovirus D68. Strain
US/MO/14-18947
Yeast	Cryptococcus neoformans1	WM629 [CBS 10079]
Yeast	Cryptococcus gattii1	Serotype B strain R272,
type VGIIb
Off-Panel
Virus	Adenovirus A12	Huie
Virus	Adenovirus C2	Adenoid 6 (NIAID 202-
001-014)
Virus	Adenovirus D20	A.A
Virus	Adenovirus E4	RI-67
Virus	Adenovirus F41	Tak
Virus	BK polyoma virus	N/A
Virus	Coronavirus 229E	229E
Virus	Coronavirus NL63	NL63 (Amsterdam I)
Virus	Coronavirus OC43	OC43
Virus	Dengue virus (Type 2)	New Guinea C
Virus	Epstein-Barr Virus	B95-8
Virus	Hepatitis B virus (HBV)	N/A
Virus	Hepatitis C virus (HCV)	N/A
Virus	Human herpes virus 7	SB
Virus	Human herpes virus 8	N/A
Virus	Human Immunodeficiency
Virus	Quantitative Synthetic
Human immunodeficiency
virus 1 (HIV-1) RNA
Virus	Human Rhinovirus A1b	2060
Virus	Human Rhinovirus A16	11757
Virus	Human Rhinovirus B3	FEB
Virus	Human Rhinovirus B83	Baylor 7 [V-190-001-021]
Virus	Influenza A H1N1	A/Florida/3/2006
Type	Pathogen
Off-Panel	Strain/Serotype
Virus	Influenza A H1N1-2009	A/California/08/2009
(H1N1pdm)
Virus	Influenza A H3N2	A/Port Chalmers/1/73
Virus	Influenza B	B/Virginia/ATCC4/2009
Virus	JC polyoma virus	MAD-4
Virus	Measles Virus	Edmonston
Virus	Mumps Virus	Jones
Virus	West Nile Virus	1986
Virus	Parainfluenza virus 2	Greer
Virus	Parainfluenza virus 4	N/A
Virus	Parvovirus B19	B19
Virus	Respiratory Syncytial Virus	A2
Virus	Rotavirus	RRV (Rhesus Rotavirus)
Virus	Rubella Virus	N/A
Virus	St. Louis Encephalitis
Virus 1	Parton
Virus	Cytomegalovirus	Davis
Virus	Herpes simplex virus 1	Macintyre
Virus	Herpes simplex virus 2	HSV-2. (Strain: MS)
Virus	Human herpes virus 6	HHV-6B. (Strain: Z29)
Virus	Human parechovirus	Serotype 3
Virus	Varicella-zoster virus	Ellen
Fungi/parasite	Candida glabrata	CBS 138
Fungi/parasite	Candida krusei	N/A
Fungi/parasite	Candida lusitaniae	Z010
Fungi/parasite	Candida metapsilosis	MCO429
Fungi/parasite	Candida orthopsilosis	MCO471
Fungi/parasite	Candida viswanathii	PK 233 [NCYC 997,
pK233]
Fungi/parasite	Candida parapsilosis	CBS 604
Fungi/parasite	Candida tropicalis	Vitek #8935
Fungi/parasite	Cryptococcus albidus	AmMS 228
Fungi/parasite	Cryptococcus amylolentus	NRRY Y-7784
Fungi/parasite	Cryptococcus laurentii	CBS 139
Fungi/parasite	Cryptococcus uniguttulatus	AmMS 234
Fungi/parasite	Cryptococcus adeliensis =
Cryptococcus adeliae =
Naganishia adeliensis	Cryptococcus adeliae
Fungi/parasite	Cryptococcus flavescens =
Papiliotrema flavescens	Cryptococcus laurentii var.
Flavescens (Saito)
Type	Pathogen	Strain/Serotype
Off-Panel
Fungi/parasite	Cryptococcus wingfieldii =
Tsuchiyaea wingfieldii	OTU 26
Fungi/parasite	Cryptococcus depauperatus
= Aspergillus depauperatus
= Filobasidiella
depauperate	K [ARSEF 2058, CBS
7842]
Fungi/parasite	Filobasidium capsuligenum	ML-186
Fungi/parasite	Naegleria fowleri	Genomic DNA
from Naegleria fowleri
Fungi/parasite	Toxoplasma gondii	Haplogroup 2
Fungi/parasite	Aspergillus fumigatus	Z014
Fungi/parasite	Candida albicans	CBS 562
Fungi/parasite	Candida dubliniensis	Z145
Bacteria	Bacillus cereus	Z091
Bacteria	Citrobacter freundii	[ATCC 13316, NCTC
9750]
Bacteria	Corynebacterium striatum	CDC F6683
Bacteria	Corynebacterium
urealyticus	3 [Garcia strain]
Bacteria	Cronobacter
(Enterobacter) sakazakii	CDC 4562-70
Bacteria	Enterobacter aerogenes	Z052
Bacteria	Enterobacter cloacae	CDC 442-68
Bacteria	Escherichia coli (non-K1)	2003-3055
Bacteria	Escherichia fergusonii	Z302
Bacteria	Escherichia hermannii	CDC 980-72
Bacteria	Escherichia vulneris	CDC 875-72
Bacteria	Haemophilus ducreyi	DCC1476 [Sweden 15A-
25]
Bacteria	Haemophilus haemolyticus	NCTC 10659
Bacteria	Haemophilus
parahaemolyticus	536 [NCTC 8479]
Bacteria	Haemophilus
parainfluenzae	NCTC 7857
Bacteria	Klebsiella pneumoniae	NCTC 9633 [NCDC 298-
53,
NCDC 410-68]
Bacteria	Listeria innocua	SLCC 3379
Bacteria	Listeria ivanovii	Li 1979
Bacteria	Morganella morganii	AM-15
Bacteria	Streptococcus salivarius	C699
Bacteria	Streptococcus sanguinis	DSS-10
Type	Pathogen	Strain/Serotype
Off-Panel
Bacteria	Streptococcus
pseudopneumoniae	CDC-SS-1757
Bacteria	Mycoplasma genitalium	M30
Bacteria	Neisseria lactamica	NCDC A7515
Bacteria	Neisseria mucosa	AmMS 138
Bacteria	Neisseria sicca	AMC 14-D-1
Bacteria	Neisseria gonorrhoeae	Z017
Bacteria	Pantoea agglomerans	Enterobacter agglomerans
Bacteria	Proprionibacterium acnes	NCTC 737
Bacteria	Proteus mirabilis	LRA 08 01 73 [API SA,
DSM 6674]
Bacteria	Pseudomonas aeruginosa	PRD-10 [CIP 103467,
NCIB 10421, PCI 812]
Yeast	Saccharomyces cerevisiae	NRRL Y-567
Bacteria	Salmonella bongori	CIP 82.33
Bacteria	Salmonella enterica	CDC K-1891
[ATCC 25928]
Bacteria	Serratia marcescens	PCI 1107
Bacteria	Shigella boydii	CDC C-123
Bacteria	Shigella flexneri	Z046
Bacteria	Shigella sonnei	AMC 43-GG9
Bacteria	Staphylococcus aureus	FDA 209
Bacteria	Staphylococcus capitis	PRA 360 677
Bacteria	Staphylococcus epidermidis	FDA strain PCI 1200
Bacteria	Staphylococcus
haemolyticus	SM 131
Bacteria	Staphylococcus hominis	Z031
Bacteria	Staphylococcus lugdunensis	LRA 260.05.79
Bacteria	Staphylococcus
saprophyticus	NCTC 7292
Bacteria	Streptococcus anginosus	NCTC 10713
Bacteria	Streptococcus bovis	Z167
Bacteria	Streptococcus dysgalactiae	Grouping strain C74
Bacteria	Streptococcus intermedius	Z126
Bacteria	Streptococcus oralis	Z307
Bacteria	Streptococcus mitis
(tigurinus)	Clinical Isolate
Bacteria	Streptococcus mutans	LRA 28.02.81

19

Page 16 of 35

20

QIAGEN QI KOLE :

QIAstat-Dx® Meningitis/Encephalitis (ME) Panel
510(k) Summary

Page 17 of 35

21

Page 18 of 35

1 Enterovirus A, B, C, D and Cryptococcus neoformans/gattii are detected by their specific rtPCR assay with no species discrimination.

2 N. meningitidis was tested at 1.00E+05 CFU/mL instead of 1.00E+06 CFU/mL.

22

| QIAstat-Dx ME Panel Target | Potential cross-reactive organism | Cross reactive
concentration |
|---------------------------------------|----------------------------------------------------------|---------------------------------|
| Haemophilus influenzae | Haemophilus haemolyticus | ≥1.00E+03 CFU/mL |
| Cryptococcus neoformans/gattii | Cryptococcus wingfieldii = Tsuchiyaea wingfieldii | ≥1.00E+01 CFU/mL |
| | Cryptococcus flavescens = Papiliotrema flavescens | ≥4.00E+03 CFU/mL |
| Cryptococcus neoformans/gattii | Cryptococcus amylolentus | ≥1.00E+01 CFU/mL |

Table 7: Samples showing cross-reactivity with the panel

In silico analysis

In silico analysis was performed, for the primer/probe designs included in the QIAstat-Dx ME Panel, in two steps to further characterize the sequence specificity of the rtPCR assays included in the QIAstat-Dx ME Panel to detect possible unspecific homologies and/or unspecific cross-reactions.

The result of the analysis performed for the primer/probe designs included in the QIAstat-Dx ME Panel pointed at 6 potential cross-reactions with Off-Panel targets (listed in Table 8).

	Table 8: Samples showing cross-reactivity with the panel

Off-Panel organism	On-Panel signal
Streptococcus pseudopneumoniae *	Streptococcus pneumoniae
Listeria innocua *	Listeria monocytogenes
Haemophilus haemolyticus	Haemophilus influenzae
Cryptococcus amylolentus	Cryptococcus
neoformans/gatti
Cryptococcus depauperatus *
Cryptococcus wingfieldii

in silico cross-reactive risk was not confirmed by in vitro testing.

Interfering Substances

The effect of potentially interfering substances on the detectability of the QIAstat-Dx ME Panel organisms was evaluated. The substances tested in the study included endogenous as well as exogenous substances that are commonly found and/or introduced into CSF specimens during specimen collection.

The QIAstat-Dx ME Panel target organisms were tested at 3x LoD in artificial CSF matrix and testing was performed in triplicate. Potential interfering substances were spiked into

23

the samples at a level predicted to be above the concentration of the substance likely to be found in CSF sample.

Potentially interfering endogenous and exogenous substances have been evaluated and have been confirmed not to interfere with any of the panel target assays at concentrations potentially found in clinical samples (above the concentration of the substance likely to be found in a real CSF specimen). As an exception, bleach was shown to cause inhibition at high concentrations (1% v/v, 0.1% v/v), and showed no interference when tested at 0.01% v/v. Results are provided in Table 9.

Table 9: Final highest concentrations per potentially interfering substance without
observable interference
Substance	Concentration
Tested	Result
Endogenous substances
Human Blood	10% (v/v)	No interference
gDNA	20 µg/mL	No interference
D(+)Glucose	10 mg/mL	No interference
L-lactate (Na)	2.2 mg/mL	No interference
Immunoglobulin G (human)	20 mg/mL	No interference
Albumin (human)	30 mg/mL	No interference
Peripheral blood mononuclear cells	10,000 cells/µL	No interference
Exogenous substances
Chlorhexidine	0.4% (w/v)	No interference
Ethanol	7.0% (v/v)	No interference
Bleach	1.0% (v/v)	Interference
Bleach	0.1% (v/v)	Interference
Bleach	0.01% (v/v)	No interference
Acyclovir	69 µg/mL	No interference
Amphotericin B	5.1 µg/mL	No interference
Ampicillin	210 µg/mL	No interference
Ceftriaxone	840 µg/mL	No interference
Cefotaxime	645 µg/mL	No interference

Microbial Interference

A microbial interference study was conducted to assess the inhibitory effects of select nontarget organisms on the ability to detect the QIAstat-Dx ME Panel target organisms. Challenging concentrations (105 units/ml for viral targets and 106 CFU/mL for bacterial targets) of non-target organisms were individually mixed with artificial CSF matrix containing spiked targeted QIAstat-Dx ME Panel organisms at 3x LoD. Testing was performed in triplicate. All QIAstat-Dx ME Panel organisms were successfully detected

24

(100% hit rate) when tested in combination with the potentially microbial interferents. See Table 10 for a list of the non-target organisms tested and the result summary.

| Substance | Supplier / Catalog # | Concentration
Tested | Result |
|---------------------------------|---------------------------|-------------------------|-----------------|
| Epstein-Barr virus | ZeptoMetrix,
0810008CF | $1E+05$ cp/mL | No interference |
| Influenza A H1N1- 2009 | ATCC, VR-1895 | $1E+05$ CEID50/mL | No interference |
| Cutibacterium acnes | ATCC, 6919 | $1E+06$ CFU/mL | No interference |
| Staphylococcus epi-
dermidis | ATCC, 14990 | $1E+06$ CFU/mL | No interference |
| Escherichia coli (non-K1) | ATCC, 25922 | $1E+06$ CFU/mL | No interference |
| Staphylococcus aureus | ATCC, 29213 | $1E+06$ CFU/mL | No interference |
| Measles Virus | ATCC, VR-24 | $1E+05$ TCID50/mL | No interference |

Table 10: Final highest concentration without observable inhibitory effect

Competitive Inhibition

Combined samples containing a mixture of two different targets spiked at low and high concentrations into artificial CSF were tested. Selection of bacteria, virus, and yeast pathogens and combinations of targets tested was based on clinical relevance.

Clinically relevant co-infection testing demonstrated that when at least two QIAstat-Dx ME Panel pathogens of different concentrations are simultaneously present in one sample, all targets can be detected by the assay. A summary of the final co-infection mixes whereby the High Positive Analyte (HPA) does not inhibit the Low Positive Analyte (LPA) is shown in Table 11.

LPA		HPA
Pathogen	Concentration	Units	Pathogen	Concentration	Units
Escherichia coli K1	3.30E+02	CFU/mL	Haemophilus influenzae	1.00E+06	CFU/mL
Haemophilus
influenzae	9.48E+02	CFU/mL	Escherichia coli K1	1.00E+06	CFU/mL
Haemophilus
influenzae	9.48E+02	CFU/mL	Streptococcus pneu-
moniae	1.00E+06	CFU/mL
Streptococcus pneu-
moniae	6.78E+02	CFU/mL	Haemophilus influenzae	1.00E+06	CFU/mL
Listeria
monocytogenes	5.58E+03	CFU/mL	Streptococcus pneu-
moniae	1.00E+06	CFU/mL
Streptococcus pneu-
moniae	6.78E+02	CFU/mL	Listeria monocytogenes	1.00E+06	CFU/mL
Cryptococcus neo-
formans	6.63E+03	CFU/mL	Streptococcus pneu-
moniae	1.00E+06	CFU/mL
Streptococcus pneu-
moniae	6.78E+02	CFU/mL	Cryptococcus neo-
formans	1.00E+05	CFU/mL
Neisseria meningitidis	3.99E+01	CFU/mL	Haemophilus influenzae	1.00E+06	CFU/mL

Table 11: Summary of competitive inhibition testing results

25

LPA			HPA
Pathogen	Concentration	Units	Pathogen	Concentration	Units
Enterovirus	4.80E+02	TCID50/
mL	Streptococcus
pyogenes	1.00E+06	CFU/mL
Streptococcus
pyogenes	1.71E+03	CFU/mL	Enterovirus	1.00E+05	TCID50/
mL

Carrvover

A carryover study was performed to evaluate the potential occurrence of crosscontamination between consecutive runs when using the QIAstat-Dx ME Panel on the QIAstat-Dx Analyzer 1.0. Pathogenic CSF samples with alternating high-positive (10-10° organism/mL) and negative samples, were conducted on two OIAstat-Dx Analyzer 1.0 instruments.

No carryover between samples was observed in the OIAstat-Dx ME Panel, demonstrating that the system design and recommended sample handling and testing practices are effective in preventing unexpected results due to carryover or cross-contamination between samples.

Sample Stability

Sample stability testing demonstrated that the OlAstat-Dx ME Panel assay is capable of processing samples (clinical cerebrospinal fluid specimens) that are stored at different temperatures without affecting the performance.

Sample stability testing demonstrated that CSF specimens may be stored at the conditions listed below.

Room temperature up to 24 hours at 15-25℃ -
-Refrigerated up to 7 days at 2-8℃

Matrix Equivalency

A matrix equivalency study was conducted to compare the performance of analytical samples prepared in negative clinical CSF matrix to samples prepared in artificial CSF matrix.

A total of 26 pathogen strains (at least two different strains per panel target) were tested in combined samples spiked in clinical true-negative CSF. Clinical CSF was sourced from commercial suppliers and was tested prior to its use in mix manufacture for true-negativity with either the QIAstat-Dx ME Panel or an alternative method.

Samples were prepared by spiking pathogens in combined (multiple-spike) sample mixes containing up to 4 organisms per sample in clinical CSF at 5x LoD, 1x LoD and 0.1x LoD concentrations following the Limit of Detection in Combined Samples study. The

26

concentration of the pathogen strain that achieved a result around the detection limit (1x LoD) was assessed by testing a minimum of 30 replicates; 5x LoD and 0.1x LoD concentrations were assessed by testing a minimum of 10 replicates. Some pathogens required an additional round of testing to establish the dilution to achieve LoD, in addition to the initial testing described.

Testing for each concentration was executed during at least 3 different days by different operators using four different lots of QIAstat-Dx ME Panel cartridges executed on 3 or more QIAstat-Dx Analyzers.

Negative samples consisted of unspiked clinical CSF, and a minimum of 10 replicates were tested.

The LoD in clinical sample matrix using combined samples has been established for 26 pathogen strains, and in 18 of those strains the LoD concentration was found to be equivalent to the one established in artificial CSF matrix. Eight (8) pathogen strains were not equivalent and had a new LoD concentration determined in clinical CSF.

Claimed LoD concentrations represent the highest (most concentrated) titer of analyte identified from LoD determination in clinical CSF or artificial CSF.

Precision (Repeatability and Reproducibility)

For the reproducibility assessment, a multi-site scheme was followed by testing both negative and positive samples at three different study sites with varying workflow variables, such as sites, days, instruments, operators and cartridge lots that could have an impact on the precision of the system. Negative samples consisted of artificial CSF. Positive combined samples consisted of artificial CSF spiked with a representative panel of pathogens covering all types of organisms targeted by the QIAstat-Dx ME Panel (i.e., RNA virus, gram (+) bacteria, gram (-) bacteria and yeast) at the limit of detection (1x LoD) and at 3x LoD. For each site, testing was performed across 5 non-consecutive days per mix with 6 replicates per day per mix (leading to a total of 90 replicates per target, concentration, and site), a minimum of 9 different QIAstat-Dx Analyzers per site, and at least 3 operators on each testing day.

Reproducibility testing was designed to evaluate the critical variables that may impact the performance of the QIAstat-Dx ME Panel in the context of its routine and intended use.

For the repeatability study, the same sample panel was tested following a single-site scheme. Repeatability testing was designed to evaluate the precision of a OIAstat-Dx ME Panel Cartridge under similar (intra laboratory) conditions. The Repeatability study was assessed with the same samples used for Reproducibility testing using Site 1.

The Reproducibility and Repeatability studies for the OIAstat-Dx ME Panel demonstrated that no test condition has a specific impact on the test performance or pathogen calling and

27

confirmed its successful precision since the panel performance or pathogen calling showed to be repeatable along the testing days (results are shown in Table 12 and Table 13).

| Grouping Variable(s) | Confidence Limit | |
|--------------------------------| | Target (%) | Lower | Cryptococcus neoformans/gattii | 1xLoD | | | | | | | Cryptococcus neoformans/gattii | 3xLoD | | | | | | | Enterovirus | | | | | | | Enterovirus | | | | | | | Escherichia coli K1 | | | | | | | Escherichia coli K1 | | | | | | | Listeria monocytogenes | | | | | | | Listeria monocytogenes | | | | | | | | Proportion | | Two-Sided 95%
---------------|------|------------|-------------------|-----------------------------------|--------|
| Concentration | Site | Fraction | Percentage
| Upper |
| 1 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 2 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 3 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| All | 90 / 90 | 100.00 | 95.98 | 100.00 |
| 1 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 2 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 3 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| All | 90 / 90 | 100.00 | 95.98 | 100.00 |
| 1xLoD | 1 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 2 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 3 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| All | 90 / 90 | 100.00 | 95.98 | 100.00 |
| 3xLoD | 1 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 2 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 3 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| All | 90 / 90 | 100.00 | 95.98 | 100.00 |
| 1xLoD | 1 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 2 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 3 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| All | 90 / 90 | 100.00 | 95.98 | 100.00 |
| 3xLoD | 1 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 2 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 3 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| All | 90 / 90 | 100.00 | 95.98 | 100.00 |
| 1xLoD | 1 | 29 / 30 | 96.67 | 82.78 | 99.92 |
| 2 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 3 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| All | 89 / 90 | 98.89 | 93.96 | 99.97 |
| 3xLoD | 1 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 2 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| 3 | 30 / 30 | 100.00 | 88.43 | 100.00 |
| All | 90 / 90 | 100.00 | 95.98 | 100.00 |

Table 12: Reproducibility study agreement rate (detected vs. expected) per analyte and the 2-sided 95% Confidence Interval by target for 1x LoD, 3x LoD and Negative samples

28

| Grouping Variable(s) | | | Proportion | | | Two-Sided 95%
Confidence Limit | |
|--------------------------|---------------|------|------------|-------------------|--|-----------------------------------|--------|
| Target | Concentration | Site | Fraction | Percentage
(%) | | Lower | Upper |
| Streptococcus agalactiae | 1xLoD | 1 | 30 / 30 | 100.00 | | 88.43 | 100.00 |
| | | 2 | 30 / 30 | 100.00 | | 88.43 | 100.00 |
| | | 3 | 30 / 30 | 100.00 | | 88.43 | 100.00 |
| | | All | 90 / 90 | 100.00 | | 95.98 | 100.00 |
| Streptococcus agalactiae | 3xLoD | 1 | 30 / 30 | 100.00 | | 88.43 | 100.00 |
| | | 2 | 30 / 30 | 100.00 | | 88.43 | 100.00 |
| | | 3 | 30 / 30 | 100.00 | | 88.43 | 100.00 |
| | | All | 90 / 90 | 100.00 | | 95.98 | 100.00 |
| n/a | Negative | 1 | 30 / 30 | 100.00 | | 88.43 | 100.00 |
| | | 2 | 30 / 30 | 100.00 | | 88.43 | 100.00 |
| | | 3 | 30 / 30 | 100.00 | | 88.43 | 100.00 |
| | | All | 90 / 90 | 100.00 | | 95.98 | 100.00 |

Table 13: Repeatability study agreement rate (detected vs. expected) per analyte and the 2-sided 95% Confidence Interval by target for 1x LoD, 3x LoD and Negative samples

| Grouping Variable(s) | | Proportion | Two-Sided 95%
Confidence Limit | | |
|--------------------------------|---------------|------------|-----------------------------------|-------|--------|
| Target | Concentration | Fraction | Percentage
(%) | Lower | Upper |
| Cryptococcus neoformans/gattii | 1xLoD | 60 / 60 | 100.00 | 94.04 | 100.00 |
| Cryptococcus neoformans/gattii | 3xLoD | 60 / 60 | 100.00 | 94.04 | 100.00 |
| Enterovirus | 1xLoD | 57 / 60 | 95.00 | 86.08 | 98.96 |
| Enterovirus | 3xLoD | 60 / 60 | 100.00 | 94.04 | 100.00 |
| Escherichia coli K1 | 1xLoD | 56 / 60 | 93.33 | 83.80 | 98.15 |
| Escherichia coli K1 | 3xLoD | 60 / 60 | 100.00 | 94.04 | 100.00 |
| Listeria monocytogenes | 1xLoD | 57 / 60 | 95.00 | 86.08 | 98.96 |
| Listeria monocytogenes | 3xLoD | 59 / 60 | 98.33 | 91.06 | 99.96 |
| Streptococcus agalactiae | 1xLoD | 60 / 60 | 100.00 | 94.04 | 100.00 |
| Streptococcus agalactiae | 3xLoD | 60 / 60 | 100.00 | 94.04 | 100.00 |
| Negative | Negative | 60 / 60 | 100.00 | 94.04 | 100.00 |

Performance Characteristics - Clinical Studies

Expected values

In the prospective clinical performance evaluation of the QIAstat-Dx Meningitis / Encephalitis (ME) Panel residual cerebrospinal fluid specimens were collected. Specimens

29

were tested at 13 geographically diverse clinical sites across 4 countries (3 sites in Europe and 10 sites in USA) from March 2022 to March 2023. The number and percentage of positive results as determined by the QIAstat-Dx ME Panel, stratified by age group are presented in Table 14. Overall. 1527 specimens were included in the prevalence assessment and the QIAstat-Dx ME Panel detected at least one organism in a total of 65 prospective specimens analysed in the study (4.3% positivity rate).

			1-17	18-44	45-64	65-84	>85
Pathogen	Overall	85 years old	31	2.0

Table 15: Demographic data for prospective evaluated specimens

31

Variable	Subgroup	N	%
Gender	Unknown	2	0.1
	Female	769	50.5
	Male	754	49.5
	Not available	1	0.1

Residual CSF specimens were tested with the OlAstat-Dx ME Panel and two types of comparator methods (an FDA-cleared molecular comparator or two validated end point PCRs followed by bidirectional sequencing (BDS) for selected targets). All targets were compared to the FDA-cleared molecular method except Streptococcus pneumoniae and Streptococcus pvogenes which were compared against two validated end point PCRs followed by bidirectional sequencing. The standard of care (SoC) testing varied across all sites but included bacterial culture, Laboratory Developed PCR test (LDT), FDA-cleared molecular methods and Cryptococcus antigen screen and culture. Standard of care (SoC) culture results were collected to allow an assessment of clinical sensitivity and specificity and were investigated in cases of discordant result.

Clinical sensitivity or positive percent agreement (PPA) was calculated as 100% x (TP / (TP + FN)). True positive (TP) indicates that both QIAstat-Dx ME Panel and comparator method have a positive result for the specific pathogen. False negative (FN) indicates that the OIAstat-Dx result is negative while the comparator result is positive for the specific pathogen. Clinical Specificity or Negative Percent agreement (NPA) was calculated as 100% x (TN / (TN + FP)). True negative (TN) indicates that both the OIAstat- Dx Panel and the comparator method have negative results for the specific pathogen. False positive (FP) indicates that the OIAstat- Dx Panel result is positive for the specific pathogen, but the comparator result is negative. The two-sided 95% confidence intervals were calculated (Wilson Score Method).

The QIAstat-Dx ME Panel prospective data for positive percent agreement (PPA) and negative percent agreement (NPA) against the comparator methods are presented by analyte in Table 16. Discrepancies between the QIAstat-Dx ME Panel and the comparator methods were investigated and discrepancy investigations are footnoted in Table 16 below.

		Positive Percent Agreement			Negative Percent Agreement
Pathogen	Sample
Category	TP/TP+FN	%	95% CI*	TN/TN+FP	%	95% CI*
Bacteria
Escherichia
coli K1a	Prospective
Fresh	2/3	66.7	20.8-93.9	969 / 969	100.0	99.6-100.0
	Prospective
Frozen	0 / 1	0.0	0.0-79.3	551 / 551	100.0	99.3-100.0
	Overall	2 / 4	50.0	15.0-85.0	1520 / 1520	100.0	99.7-100.0

Table 16: Prospectively Collected Clinical Specimens (fresh and frozen) Performance Agreement (PPA and NPA) between QIAstat-Dx ME Panel and Comparator Method by Target

32

QIAGEN

QIAstat-Dx® Meningitis/Encephalitis (ME) Panel
510(k) Summary

	Page 29 of 35

		Positive Percent Agreement		Negative Percent Agreement
Pathogen	Sample
Category	TP/TP+FN	%	95% CI*	TN/TN+FP	%	95% CI*
Bacteria
Haemophilus
influenzae b	Prospective
Fresh	0 / 1	0.0	0.0-79.3	970 / 971	99.9	99.4-100.0
	Prospective
Frozen	4 / 4	100.0	51.0-100.0	546 / 548	99.6	98.7-99.9
	Overall	4 / 5	80.0	37.6-96.4	1516 / 1519	99.8	99.4-99.9
Listeria
monocytogenes c	Prospective
Fresh	1 / 1	100.0	20.7-100.0	971 / 971	100.0	99.6-100.0
	Prospective
Frozen	3 / 4	75.0	30.1-95.4	548 / 548	100.0	99.3-100.0
	Overall	4 / 5	80.0	37.6-96.4	1519 / 1519	100.0	99.7-100.0
Neisseria
meningitidis
(encapsulated)d	Prospective
Fresh	1 / 1	100.0	20.7-100.0	971 / 971	100.0	99.6-100.0
	Prospective
Frozen	0 / 0	N/A	N/A	551 / 552	99.8	99.0-100.0
	Overall	1 / 1	100.0	20.7-100.0	1522 / 1523	99.9	99.6-100.0
Streptococcus
agalactiae	Prospective
Fresh	2 / 2	100.0	34.2-100.0	970 / 970	100.0	99.6-100.0
	Prospective
Frozen	1 / 1	100.0	20.7-100.0	551 / 551	100.0	99.3-100.0
	Overall	3 / 3	100.0	43.9-100.0	1521 / 1521	100.0	99.7-100.0
Streptococcus
pneumoniae e	Prospective
Fresh	1 / 1	100.0	20.7-100.0	845 / 848	99.6	99.0-99.9
	Prospective
Frozen	7 / 7	100.0	64.6-100.0	515 / 517	99.6	98.6-99.9
	Overall	8 / 8	100.0	67.6-100.0	1360 / 1365	99.6	99.1-99.8
Streptococcus
pyogenes	Prospective
Fresh	0 / 0	N/A	N/A	778 / 778	100.0	99.5-100.0
	Prospective
Frozen	0 / 0	N/A	N/A	513 / 513	100.0	99.3-100.0
	Overall	0 / 0	N/A	N/A	1291 / 1291	100.0	99.7-100.0
Virus
Enterovirus
(EV)f	Prospective
Fresh	18 / 20	90.0	69.9-97.2	951 / 952	99.9	99.4-100.0
	Prospective
Frozen	4 / 4	100.0	51.0-100.0	548 / 548	100.0	99.3-100.0
	Overall	22 / 24	91.7	74.2-97.7	1499 / 1500	99.9	99.6-100.0
Fungi / Yeast
Cryptococcus
gattii /
Cryptococcus
neoformans
(not
differentiated)g	Prospective
Fresh	2 / 5	40.0	11.8-76.9	965 / 967	99.8	99.2-99.9
	Prospective
Frozen	2 / 2	100.0	34.2-100.0	550 / 550	100.0	99.3-100.0
	Overall	4 / 7	57.1	25.0-84.2	1515 / 1517	99.9	99.5-100.0

33

ª For the prospective fresh Escherichia coli KI discordant sample, no organisms were detected with resolution method PCR/BDS. For the frozen Escherichia coli K1 discordant sample no organisms were detected with bacterial culture.

b For the prospective fresh Haemophilus influenzae discordant sample, no organisms were detected by the SoC bacterial culture and resolution testing with PCR/BDS. Of the three (3) false positive Haemophilus influenzae samples, no organisms were detected in one frozen by SoC culture and PCR/BDS resolution method was also negative. No additional testing results associated with the final frozen false positive sample were available.

6 For the frozen Listeria monocytogenes discordant sample the negative result was confirmed positive with SoC culture and LDT result was positive.

d For the frozen Neisseria meningitidis (encapsulated) sample, no organisms were detected by SoC culture and LDT resolution testing with PCR/BDS also returned a negative result for this sample.

e Of the five (5) false positive Streptococcus pneumoniae samples, no organisms were detected in four (3 fresh and 1 frozen) prospective samples with SoC culture. One prospective frozen sample had no SoC result available. However, an FDA cleared method conducted as part of the study also produced a negative result.

f For the prospective fresh Enterovirus discordant samples, no organisms were detected in one sample by two independent SoC LDT assays. The negative result for the second sample returned a negative result with PCR/BDS. For the prospective fresh false positive Enterovirus sample, a negative result was returned when tested with PCR/BDS.

8 Of the three false negative Cryptococcus neoformans (not differentiated) results, no organisms were detected in two fresh samples with fungal culture and PCR/BDS. The remaining false negative fresh sample was confirmed negative for Cryptococcus gattii / Cryptococcus neoformans (not differentiated) with PCR/BDS. Of the two false positive results, no organisms were detected for one fresh sample with PCR/BDS. No organisms were detected in the second fresh sample with SoC fungal culture.

Archived Specimens Testing

Several analytes were either not encountered or had a low prevalence in the prospective arm of the study, so an effort was made to collect and test as many archived samples (positive targeted archived specimens) as possible for all targets within the panel. A total of 195 retrospective archived specimens were enrolled into the study. One hundred and fifty-four (154) archived specimens were excluded from the analysis as positivity was not confirmed by the comparator method. A total of 41 evaluable archived specimens were used in the analysis to support the QIAstat-Dx ME Panel performance evaluation and Table 17 provides a summary of demographic information for the archived specimens.

Table 17: Demographic Summary of Evaluable Archived Specimens for QIAstat-Dx ME Panel Clinical Evaluation

Sample Category	Variable	Subgroup	N	%
Archived	Age Group	Cryptococcus
gattii	Archived	2 / 2	100.0	34.2-100.0
	Prospective	1 / 1	100.0	20.7-100.0
Cryptococcus
neoformans
(not
differentiated)	Fresh
	Prospective
Frozen	0 / 0	N/A	N/A	24 / 24

Table 20: Fungal Culture Comparison for diagnostic sensitivity and specificity by sample category

Validity of results

The failure rate for initial clinical specimens tests for prospective fresh was 2.7% (26/977), for prospective frozen was 1.3% (7/555) for prospective frozen and for archived samples was 1.7% (3/176). Those failures consisted of 5 instrument errors and 2 invalid results, and 29 were other run failures. All specimens except 5 (3 prospective fresh and 2 prospective frozen) were retested and were successful after retest, yielding a final success rate of 99.7% for prospective fresh, 99.6% for prospective frozen and 100.0% for archived samples. The failure rates breakdown due to instrument, invalid results and other run failures is summarized in Table 21. Withdrawn specimens were not included in the validity assessment; however some prospective samples which were excluded from the agreement analysis were included for the validity assessment (3 prospective fresh, 5 prospective frozen and 135 archive specimens).

Table 21: Summary of the Number of Samples with Failed Test Results (Initial and
Final)
Placeholder Column 1	Placeholder Column 2
Placeholder Data 1	Placeholder Data 2

| | | Initial Runs | | Final Runs
(after repeats) | |
|--------------------|----------------|--------------|-----|-------------------------------|-----|
| Sample Category | Failure Reason | N/Total | % | N/Total | % |
| Prospective Fresh | Invalid* | 0 / 977 | 0.0 | 0 / 977 | 0.0 |
| Prospective Fresh | Instrument | 3 / 977 | 0.3 | 0 / 977 | 0.0 |
| Prospective Fresh | Other** | 23 / 977 | 2.4 | 3 / 977 | 0.3 |
| Prospective Frozen | Invalid | 0 / 555 | 0.0 | 0 / 555 | 0.0 |
| Prospective Frozen | Instrument | 1 / 555 | 0.2 | 0 / 555 | 0.0 |
| Prospective Frozen | Other | 6 / 555 | 1.1 | 2 / 555 | 0.4 |
| Archived | Invalid | 2 / 176 | 1.1 | 0 / 176 | 0.0 |
| Archived | Instrument | 1 / 176 | 0.6 | 0 / 176 | 0.0 |
| Archived | Other | 0 / 176 | 0.0 | 0 / 176 | 0.0 |

*Internal Control failures with at least one analyte detected and the other analytes reported as 'invalid'

** Run failures related to workflow checkpoints.

37

Contrived Specimens Testing

Contrived specimens testing was conducted to support the nine (9) targets on the panel as there were insufficient positive specimens obtained from both prospective and archived collection efforts. Contrived specimens were prepared by spiking five different quantified strains representative of the genetic diversity of each pathogen. For each pathogen, the LoD concentration was manufactured at 2x (at least 50%) and 5x LoD spiked into screened individual unique samples of negative CSF. Contrived specimens were tested alongside negative specimens in a blinded manner. The results are summarized in Table 22.

Table 22: Overall Proportion of Positive Results Post Exclusions or withdrawals of Contrived Positive Samples for Each Target

| Classification
(genome type) | Pathogen | Concentration
Level | Frequency
of Positive
Results | Proportion
(%) of
Positive
Results | Lower
95%
Confidence
Limit | Upper
95%
Confidence
Limit |
|---------------------------------|-----------------------------------------------------------------------------|------------------------|-------------------------------------|---------------------------------------------|-------------------------------------|-------------------------------------|
| Bacteria | Escherichia coli Kl | 2xLoD | 48 / 48 | 100.0 | 92.6 | 100.0 |
| | | 5xLoD | 37 / 37 | 100.0 | 90.6 | 100.0 |
| | | Total | 85 / 85 | 100.0 | 95.7 | 100.0 |
| | Haemophilus influenzae | 2xLoD | 57 / 57 | 100.0 | 93.7 | 100.0 |
| | | 5xLoD | 36 / 36 | 100.0 | 90.4 | 100.0 |
| | | Total | 93 / 93 | 100.0 | 96.0 | 100.0 |
| | Listeria monocytogenes | 2xLoD | 47 / 49 | 95.9 | 86.3 | 98.9 |
| | | 5xLoD | 38 / 38 | 100.0 | 90.8 | 100.0 |
| | | Total | 85 / 87 | 97.7 | 92.0 | 99.4 |
| | Neisseria meningitidis
(encapsulated) | 2xLoD | 46 / 48 | 95.8 | 86.0 | 98.8 |
| | | 5xLoD | 39 / 40 | 97.5 | 87.1 | 99.6 |
| | | Total | 85 / 88 | 96.6 | 90.5 | 98.8 |
| | Streptococcus
agalactiae | 2xLoD | 49 / 49 | 100.0 | 92.7 | 100.0 |
| | | 5xLoD | 39 / 39 | 100.0 | 91.0 | 100.0 |
| | | Total | 88 / 88 | 100.0 | 95.8 | 100.0 |
| | Streptococcus
pneumoniae | 2xLoD | 55 / 57 | 96.5 | 88.1 | 99.0 |
| | | 5xLoD | 39 / 39 | 100.0 | 91.0 | 100.0 |
| | | Total | 94 / 96 | 97.9 | 92.7 | 99.4 |
| | Streptococcus pyogenes | 2xLoD | 47 / 49 | 95.9 | 86.3 | 98.9 |
| | | 5xLoD | 40 / 40 | 100.0 | 91.2 | 100.0 |
| | | Total | 87 / 89 | 97.8 | 92.2 | 99.4 |
| Virus | Enterovirus (EV) | 2xLoD | 48 / 49 | 98.0 | 89.3 | 99.6 |
| | | 5xLoD | 39 / 39 | 100.0 | 91.0 | 100.0 |
| | | Total | 87 / 88 | 98.9 | 93.8 | 99.8 |
| Fungi / Yeast | Cryptococcus gattii /
Cryptococcus
neoformans (not
differentiated) | 2xLoD | 41 / 41 | 100.0 | 91.4 | 100.0 |
| | | 5xLoD | 38 / 38 | 100.0 | 90.8 | 100.0 |
| | | Total | 79 / 79 | 100.0 | 95.4 | 100.0 |

38

Target Proportion of Positive Result ≥95% was achieved for all prepared contrived samples 2xLoD and 5xLoD in all tested analytes.

Conclusions

The QIAstat-Dx ME Panel is substantially equivalent to the to the predicate device, FilmArray Meningitis/Encephalitis (ME) Panel.