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.

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);
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.

AI/ML Overview

The provided document is a 510(k) Summary for the QIAGEN GmbH QIAstat-Dx Meningitis/Encephalitis (ME) Panel. Here's a breakdown of the acceptance criteria and study information:

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state formal acceptance criteria (i.e., predefined thresholds for sensitivity and specificity). However, it reports sensitivity (Positive Percent Agreement - PPA) and specificity (Negative Percent Agreement - NPA) values from its clinical performance study. The reported performance is implicitly the "met acceptance criteria" as the device received 510(k) clearance.

Here's a summary of the clinical performance for each pathogen from the prospective clinical study (Table 16), which represents the primary evidence for diagnostic performance:

Pathogen	Sample Category	PPA (TP/TP+FN) (%)	95% CI (PPA)	NPA (TN/TN+FP) (%)	95% CI (NPA)
Escherichia coli K1	Overall	50.0 (2/4)	15.0-85.0	100.0 (1520/1520)	99.7-100.0
Haemophilus influenzae	Overall	80.0 (4/5)	37.6-96.4	99.8 (1516/1519)	99.4-99.9
Listeria monocytogenes	Overall	80.0 (4/5)	37.6-96.4	100.0 (1519/1519)	99.7-100.0
Neisseria meningitidis (encapsulated)	Overall	100.0 (1/1)	20.7-100.0	99.9 (1522/1523)	99.6-100.0
Streptococcus agalactiae	Overall	100.0 (3/3)	43.9-100.0	100.0 (1521/1521)	99.7-100.0
Streptococcus pneumoniae	Overall	100.0 (8/8)	67.6-100.0	99.6 (1360/1365)	99.1-99.8
Streptococcus pyogenes	Overall	N/A (0/0)	N/A	100.0 (1291/1291)	99.7-100.0
Enterovirus (EV)	Overall	91.7 (22/24)	74.2-97.7	99.9 (1499/1500)	99.6-100.0
Cryptococcus gattii / Cryptococcus neoformans	Overall	57.1 (4/7)	25.0-84.2	99.9 (1515/1517)	99.5-100.0

Additionally, for contaminants that were confirmed by culture (fungal and bacterial), in table 19 and 20:

Pathogen	Sample Category	Sensitivity (compared to culture) (%)	95% CI (Sensitivity)	Specificity (compared to culture) (%)	95% CI (Specificity)
Bacteria
Escherichia coli Kl	Archived	100.0 (1/1)	20.7-100.0	90.9 (10/11)	62.3-98.4
	Prospective Fresh	50.0 (1/2)	9.5-90.5	100.0 (760/760)	99.5-100.0
Haemophilus influenzae	Archived	100.0 (1/1)	20.7-100.0	90.9 (10/11)	62.3-98.4
	Prospective Frozen	100.0 (3/3)	43.9-100.0	99.7 (339/340)	98.4-99.9
Listeria monocytogenes	Prospective Fresh	100.0 (1/1)	20.7-100.0	100.0 (761/761)	99.5-100.0
	Prospective Frozen	66.7 (2/3)	20.8-93.9	100.0 (340/340)	98.9-100.0
Neisseria meningitidis (encapsulated)	Archived	100.0 (2/2)	34.2-100.0	90.0 (9/10)	59.6-98.2
Streptococcus agalactiae	Prospective Fresh	100.0 (1/1)	20.7-100.0	99.9 (760/761)	99.3-99.9
	Prospective Frozen	100.0 (1/1)	20.7-100.0	100.0 (342/342)	98.9-100.0
Streptococcus pneumoniae	Prospective Frozen	100.0 (3/3)	43.9-100.0	99.7 (339/340)	98.4-99.9
Fungi / Yeast
Cryptococcus gattii / Cryptococcus neoformans	Archived	100.0 (2/2)	34.2-100.0	100.0 (1/1)	20.7-100.0
	Prospective Fresh	100.0 (1/1)	20.7-100.0	98.5 (129/131)	94.6-99.6

2. Sample Sizes Used for the Test Set and Data Provenance

Test Set (Clinical Performance Study):
- Prospective Specimens: 1524 evaluable specimens.
  - 552 (36.2%) were frozen before testing.
  - 972 (63.8%) were tested fresh.
- Archived Specimens: 41 evaluable archived specimens (from an initial 195 collected).
- Contrived Specimens: Not specified as a "test set" in the context of clinical performance, but used to supplement for rare analytes.
  - Ranges from 79 to 96 samples per pathogen, tested at 2xLoD and 5xLoD (e.g., Cryptococcus neoformans/gattii had 79 samples). These were likely individual spiked samples.
Data Provenance:
- Country of Origin: 13 geographically diverse clinical sites across 4 countries (10 U.S. sites and 3 European sites).
- Retrospective/Prospective: The study included both prospective (March 2022 to March 2023) and retrospective (archived) specimens.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications

The document does not explicitly state the number of "experts" used to establish ground truth or their specific qualifications (e.g., radiologist with X years of experience).

Instead, the ground truth for the clinical performance study was established using:

An FDA-cleared molecular comparator method.
Two validated end point PCRs followed by bidirectional sequencing (BDS) for Streptococcus pneumoniae and Streptococcus pyogenes.
Standard of Care (SoC) testing, which varied across sites and included bacterial culture, Laboratory Developed PCR tests (LDT), FDA-cleared molecular methods, and Cryptococcus antigen screen and culture.
Discrepancy investigations were conducted for discordant results, implying a review process, but details on who performed this review are not given.

4. Adjudication Method for the Test Set

The document mentions that discrepancies between the QIAstat-Dx ME Panel and the comparator methods were investigated. This implies an adjudication process was in place to determine the true positive/negative status for discordant results. However, the specific method (e.g., 2+1, 3+1, none) is not explicitly described. The footnotes in Table 16 (Clinical Performance) provide details on how some discordant cases were resolved (e.g., "no organisms were detected with resolution method PCR/BDS," "negative result was confirmed positive with SoC culture and LDT result was positive").

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study comparing human readers with and without AI assistance was not done. This device is an in vitro diagnostic (IVD) test for nucleic acid detection, not an imaging AI device that assists human readers.

6. Standalone (i.e., algorithm only without human-in-the-loop performance) Study

Yes, a standalone performance study was done. The entire clinical performance study (Prospective, Archived, and Contrived Specimens Testing) evaluates the performance of the QIAstat-Dx ME Panel (the algorithm/device) directly against comparator methods (other molecular tests, culture, PCR/BDS), without a human-in-the-loop component. The device generates results automatically, and its accuracy is assessed based on these outputs.

7. Type of Ground Truth Used

The ground truth for the clinical performance studies was established using a combination of:

FDA-cleared molecular comparator method.
Validated end point PCRs followed by bidirectional sequencing (BDS).
Standard of Care (SoC) culture (for bacterial and fungal analytes).
Laboratory-Developed PCR tests (LDT).
Discrepancy investigations where discordant results were resolved using additional testing.

For the contrived specimens, the ground truth was known by design, as the samples were intentionally spiked with quantified strains.

8. Sample Size for the Training Set

The document does not provide details about a specific "training set" sample size for the QIAstat-Dx ME Panel. As an IVD based on real-time PCR, its "training" is typically in the form of analytical validation and optimization during its development, rather than machine learning model training with a distinct training dataset. The studies described are primarily for clinical performance validation, demonstrating the device's accuracy in a real-world setting.

9. How the Ground Truth for the Training Set Was Established

Since a dedicated "training set" for a machine learning model is not described, the concept of establishing ground truth for it is not applicable here in the conventional sense of AI/ML. The analytical validation studies (Limit of Detection, Analytical Reactivity/Inclusivity, Analytical Specificity/Exclusivity, etc.) involved known concentrations and strains of pathogens (e.g., ATCC strains, commercial stocks) in artificial or negative clinical CSF. The "ground truth" for these analytical studies was based on the known composition and concentration of these prepared samples.

Summary

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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"

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(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).

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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

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Indications for Use

510(k) Number (if known) K242256

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

Indications for Use (Describe)

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)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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510(k) Summary

General Information

Submitted by:	QIAGEN GmbHQIAGEN Strasse 1Hilden, Germany 40724
Contact Person:	Sonia PabloSenior Manager, Regulatory AffairsSTAT-Dx Life, S.L. (A QIAGEN Company)Carrer Baldiri Reixac, 408028 BarcelonaSpainPhone: +34 696 85 81 85Email: 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

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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.

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);

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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.

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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	FilmArrayMeningitis/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 multiplexednucleic acid real-time PCR-basedin vitro diagnostic test intendedfor use with the QIAstat-DxAnalyzer 1.0. The QIAstat-DxME Panel is capable ofsimultaneous detection andidentification of multiplebacterial, viral, and yeast nucleicacids from cerebrospinal fluid(CSF) specimens obtained vialumbar puncture from individualswith signs and/or symptoms ofmeningitis and/or encephalitis.The following organisms areidentified using the QIAstat-DxME Panel:Bacteria:• Escherichia coli K1• Haemophilus influenzae• Listeria monocytogenes	The FilmArrayMeningitis/Encephalitis (ME)Panel is a qualitativemultiplexed nucleic acid-basedin vitro diagnostic test intendedfor use with FilmArray,FilmArray 2.0, and FilmArrayTorch systems. The FilmArrayME Panel is capable ofsimultaneous detection andidentification of multiplebacterial, viral, and yeastnucleic acids directly fromcerebrospinal fluid (CSF)specimens obtained via lumbarpuncture from individuals withsigns and/or symptoms ofmeningitis and/or encephalitis.The following organisms areidentified using the FilmArrayME Panel:Bacteria:• Escherichia coli K1• Haemophilus influenzae• Listeria monocytogenes

	Table 1: Comparison of the QIAstat-Dx ME Panel with the predicate device
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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
	• Cryptococcusneoformans/gattii*	• Human herpesvirus 6
		• Human parechovirus
		• Varicella zoster virus
	The QIAstat-Dx ME Panel isindicated as an aid in thediagnosis of specific agents ofmeningitis and/or encephalitis andresults must be used inconjunction with other clinical,epidemiological, and laboratorydata. Results from the QIAstat-Dx ME Panel are not intended tobe used as the sole basis fordiagnosis, treatment, or otherpatient management decisions.Positive results do not rule out co-infection with organisms notincluded in the QIAstat-Dx MEPanel. The agent or agentsdetected may not be the definitecause of the disease. Negativeresults do not preclude centralnervous system (CNS) infection.Not all agents of CNS infectionare detected by this test andsensitivity in clinical use may
		Yeast:
		• Cryptococcusneoformans/gattii
		The FilmArray ME Panel isindicated as an aid in thediagnosis of specific agents ofmeningitis and/or encephalitisandresults are meant to be used inconjunction with other clinical,epidemiological, and laboratorydata. Results from theFilmArray ME Panel are notintended to be used as the solebasis for diagnosis, treatment, orother patient managementdecisions. Positive results do notrule out co-infection withorganisms not included in theFilmArray ME Panel. The agentdetected may not be the definitecause of the disease. Negativeresults do not preclude centralnervous system (CNS)infection. Not all agents of CNSinfection are detected by thistest and sensitivity in clinicaluse may differ from that
Characteristic	Device	Predicate
	differ from that described in theinstructions for use.	described in the package insert.
	The QIAstat-Dx ME Panel is notintended for testing of specimenscollected from indwelling CNSmedical devices.	The FilmArray ME Panel is notintended for testing ofspecimens collected fromindwelling CNS medicaldevices.
	The QIAstat-Dx ME Panel isintended to be used in conjunctionwith standard of care culture fororganism recovery, serotyping, andantimicrobial susceptibility testing.	The FilmArray ME Panel isintended to be used inconjunction with standard ofcare culture for organismrecovery,serotyping, and antimicrobialsusceptibility testing.
	* Cryptococcus neoformans andCryptococcus gattii are notdifferentiated.
Specimen Type	Cerebrospinal Fluid	Cerebrospinal Fluid
Analyte Detected	RNA/DNA	RNA/DNA
OrganismsDetected	See above	See above
Amplification andDetectionTechnology	PCR	PCR
Differences
Assay Controls	One internal control in eachcartridge to control for sampleprocessing that is subjected to allnucleic acid extraction andamplification steps similar topatient samples. Labeling willrecommend use of negative andpositive external controlsregularly. Use transport media asthe external negative control, andpreviously characterized positivesamples or negative samplespiked with well characterizedtarget organisms as externalpositive controls.	Two controls are included ineach reagent pouch to controlfor sample processing andboth stages of PCR and meltanalysis. Labeling recommendsthe use of external positive andnegative controls regularly.Enteric transport media can beused as an external negativecontrol, and previouslycharacterized positive samplesor negative samples spiked withwell characterized organisms asexternal 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 AcidExtractionTechnology	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 andDetectionInstrumentSystem	QIAstat-Dx Analyzer 1.0	FilmArray Instruments

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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	LoDConcentration*	DetectionRate
Escherichia coli Kl	Strain C5 [Bort];018ac:K1:H7	ATCC700973	3.48E+02 CFU/mL	30/30
Escherichia coli K1	NCTC 9001.Serovar 01:K1:H7	ATCC11775	7.86E+02 CFU/mL	30/30
Haemophilus influenzae	type b (cap)	ATCC10211	3.16E+02 CFU/mL	32/32
Haemophilus influenzae	Type e [strain AMC36-A-7	ATCC8142	2.54E+03 CFU/mL	30/30
Listeria monocytogenes	Type 1/2b	ZeptoMetrix0801534	1.86E+03 CFU/mL	21/21
Listeria monocytogenes	Type 4b. Strain Li 2	ATCC19115	6.64E+03 CFU/mL	30/30
Neisseria meningitidis(encapsulated)	Serotype B. M2092	ATCC13090	8.28E-02 CFU/mL	31/32
Neisseria meningitidis(encapsulated)	Serotype Y. M-112[BO-6]	ATCC35561	1.33E+01 CFU/mL	30/30
Streptococcus agalactiae	Z019	ZeptoMetrix0801545	1.75E+03 CFU/mL	30/30
Streptococcus agalactiae	G19 group B	ATCC13813	3.38E+03 CFU/mL	31/31
Streptococcus pneumoniae	19F	ZeptoMetrix0801439	7.14E+02 CFU/mL	29/30
Streptococcus pneumoniae	Serotype 1. NCTC7465	ATCC33400	6.22E-01 CFU/mL	29/29
Streptococcus pyogenes	Z472; Serotype M1	ZeptoMetrix0804351	1.80E+03 CFU/mL	30/30
Streptococcus pyogenes	Bruno [CIP104226]	ATCC19615	9.10E+01CFU/mL	30/30
Enterovirus A	CoxsackievirusA16	ZeptoMetrix0810107CF	3.79E+00 TCID50/mL	31/31
Enterovirus A	A6, species A.Strain Gdula	ATCCVR-1801	1.60E+02 TCID50/mL	30/30
Enterovirus B	Coxsackievirus B5	ZeptoMetrix0810019CF	8.91E+01 TCID50/mL	30/30
Enterovirus B	Coxsackievirus A9,species B	ZeptoMetrix0810017CF	4.36E+01 TCID50/mL	28/29
Enterovirus C	CoxsackievirusA17, species C.Strain G-12	ATCCVR-1023	1.58E+01 TCID50/mL	30/30
Enterovirus C	CoxsackievirusA24. Starin DN-19	ATCCVR-583	4.99E+00 TCID50/mL	30/30
Pathogen	Strain	Source /Catalog ID	LoDConcentration*	DetectionRate
Enterovirus D	EV 70, species D,strain J670/71	ATCCVR-836	$4.99E+01$ TCID50/mL	30/31
Enterovirus D	Enterovirus D68.Strain US/MO/14-18947	ATCCVR-1823	$5.06E+02$ TCID50/mL	30/30
Cryptococcus neoformans	Serotype D strainWM629, typeVNIV	ATCCMYA-4567	$2.21E+03$ CFU/mL	31/31
Cryptococcus neoformans	C. neoformans H99	ATCC208821	$1.64E+02$ CFU/mL	31/31
Cryptococcus gattii	Serotype B strainR272, type VGIIb	ATCCMYA-4094	$1.32E+04$ CFU/mL	30/30
Cryptococcus gattii	A6MR38 [CBS11545]	ATCCMYA-4877	$2.60E+03$ CFU/mL	29/29

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

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*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 ConcentrationTested
Escherichiacoli 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 ConcentrationTested
Haemophilusinfluenzae	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
Listeriamonocytogenes	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 [LMG21264, 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
Neisseriameningitidis(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
Streptococcusagalactiae	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 strainD136C(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 ConcentrationTested
	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
Streptococcuspneumoniae	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
Streptococcuspyogenes	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 strainT12 [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, StrainConn-5	3x LoD
	ATCC (VR-29)	Species B, Serotype CV-B2. StrainOhio-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 ConcentrationTested
	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. StrainKuykendall [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. StrainUS/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 (09/2014Isolate 2)	1:100 from stock
	ATCC (VR-1825)	Enterovirus D68. StrainUS/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, typeVNIV	1x LoD
	ATCC (208821)a	H99	1x LoD
	ATCC (32045)	type strain, CBS 132	3x LoD
	ATCC (MYA-4564)	Serotype A strain WM148, typeVNI	1:1000 from stock
	ATCC (13690)	M2092	1:100 from stock
Cryptococcusneoformans	ATCC (MYA-4566)	Serotype AD strain WM628, typeVNIII	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, typeVGIIb	1x LoD
	ATCC (MYA-4877)a	A6MR38	1x LoD
Cryptococcusgattii	ATCC (MYA-4560)	Serotype B strain WM179, typeVGI	1:100 from stock
	ATCC (MYA-4562)	Serotype B strain WM161, typeVGIII	1:1000 from stock
	ATCC (MYA-4563)	Serotype C strain WM779, typeVGIV	1:1000 from stock
	ATCC (MYA-4138)	A1M R265	3x LoD
Pathogen	Source(Catalog ID)	Strain/Serotype	Lowest ConcentrationTested
	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

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{14}------------------------------------------------

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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 allVGI,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-B80through 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-1through PV-3).
Escherichia coli K1	K1 strains

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

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Pathogens	Clinically relevant strains/subtypes detected
Rest of On-Panel organismwith no biologicalsubclassification (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

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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. StrainUS/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 ImmunodeficiencyVirus	Quantitative SyntheticHuman immunodeficiencyvirus 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	PathogenOff-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 EncephalitisVirus 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= Filobasidielladepauperate	K [ARSEF 2058, CBS7842]
Fungi/parasite	Filobasidium capsuligenum	ML-186
Fungi/parasite	Naegleria fowleri	Genomic DNAfrom 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, NCTC9750]
Bacteria	Corynebacterium striatum	CDC F6683
Bacteria	Corynebacteriumurealyticus	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	Haemophilusparahaemolyticus	536 [NCTC 8479]
Bacteria	Haemophilusparainfluenzae	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	Streptococcuspseudopneumoniae	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	Staphylococcushaemolyticus	SM 131
Bacteria	Staphylococcus hominis	Z031
Bacteria	Staphylococcus lugdunensis	LRA 260.05.79
Bacteria	Staphylococcussaprophyticus	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

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Page 16 of 35

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QIAGEN QI KOLE :

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

Page 17 of 35

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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.

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QIAstat-Dx ME Panel Target	Potential cross-reactive organism	Cross reactiveconcentration
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	Cryptococcusneoformans/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

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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	ConcentrationTested	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

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(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 #	ConcentrationTested	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
Haemophilusinfluenzae	9.48E+02	CFU/mL	Escherichia coli K1	1.00E+06	CFU/mL
Haemophilusinfluenzae	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
Listeriamonocytogenes	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

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LPA			HPA
Pathogen	Concentration	Units	Pathogen	Concentration	Units
Enterovirus	4.80E+02	TCID50/mL	Streptococcuspyogenes	1.00E+06	CFU/mL
Streptococcuspyogenes	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

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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

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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)			Proportion		Two-Sided 95%Confidence Limit
Target	Concentration	Site	Fraction	Percentage(%)	Lower	Upper
Cryptococcus neoformans/gattii	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
Cryptococcus neoformans/gattii	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
Enterovirus	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
Enterovirus	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
Escherichia coli K1	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
Escherichia coli K1	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
Listeria monocytogenes	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
Listeria monocytogenes	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

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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

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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	<1 year	years old	years old	years old	years old	years old	Unknown
Bacteria
Escherichia coli K1	2(0.1%)	2(1.2%)	0(0.0%)	0(0.0%)	0(0.0%)	0(0.0%)	0(0.0%)	0(0.0%)
Haemophilusinfluenzae	7(0.5%)	0(0.0%)	0(0.0%)	1(0.2%)	5(1.1%)	1(0.3%)	0(0.0%)	0(0.0%)
Listeriamonocytogenes	4(0.3%)	0(0.0%)	0(0.0%)	0(0.0%)	2(0.5%)	1(0.3%)	1(3.2%)	0(0.0%)
Neisseriameningitidis(encapsulated)	2(0.1%)	0(0.0%)	0(0.0%)	1(0.2%)	1(0.2%)	0(0.0%)	0(0.0%)	0(0.0%)
Streptococcusagalactiae	3(0.2%)	1(0.6%)	0(0.0%)	0(0.0%)	2(0.5%)	0(0.0%)	0(0.0%)	0(0.0%)
Streptococcuspneumoniae	17(1.1%)	3(1.8%)	0(0.0%)	3(0.7%)	7(1.6%)	4(1.2%)	0(0.0%)	0(0.0%)
Streptococcuspyogenes	1(0.1%)	0(0.0%)	1(0.8%)	0(0.0%)	0(0.0%)	0(0.0%)	0(0.0%)	0(0.0%)
Virus
Enterovirus (EV)	23(1.5%)	12(7.4%)	3(2.3%)	5(1.2%)	1(0.2%)	1(0.3%)	0(0.0%)	1(50.0%)
Fungi / Yeast
Cryptococcus gattii /Cryptococcusneoformans(not differentiated)	6(0.4%)	0(0.0%)	0(0.0%)	1(0.2%)	5(1.1%)	0(0.0%)	0(0.0%)	0(0.0%)
Overall Panel Result
Negative	1462(95.7%)	145(89.0%)	124(96.9%)	407(97.4%)	419(94.8%)	336(98.0%)	30(96.8%)	1(50.0%)
Positive	65(4.3%)	18(11.0%)	4(3.1%)	11(2.6%)	23(5.2%)	7(2.0%)	1(3.2%)	1(50.0%)

Table 14: Table Summary of Prevalence of Target Pathogens, as identified by QIAstat-Dx ME Panel, stratified by age

N.B. Total of positive + negative specimens is 1527 due to inclusion of 3 specimens where a QIAstat-Dx result was available but no FilmArray result was available (therefore classed as non-evaluable for the performance analysis).

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Clinical Performance

The performance characteristics of the QIAstat-Dx ME Panel was assessed by a multicentre, observational, prospective, and retrospective, clinical performance study, testing fresh and frozen cerebrospinal fluid (CSF) residual specimens obtained by lumbar puncture from patients with signs and symptoms of meningitis and/or encephalitis. The study was conducted at 13 geographically diverse study sites: ten (10) U.S. sites and three (3) European sites. Testing of prospectively collected residual specimens occurred between March 2022 and March 2023. All study sites were hospital-associated or independent clinical diagnostics laboratories that perform routine diagnostics.

Prospective Specimens Testing

A total of 1737 prospective residual CSF specimens were enrolled into the clinical study, of which 205 were withdrawn. The most common reason for specimen withdrawal was ineligibility. Additionally, some prospective samples could not be included in the agreement analysis due to missing data. The final dataset consisted of 1524 prospective specimens of which 552 (36.2%) were frozen before testing and 972 (63.8%) were tested fresh.

Table 15 provides a summary of demographic information for the 1524 specimens included in the prospective study.

Sample Category	Variable	Subgroup	N	%
Prospective Fresh	Age Group	<1 year	136	14.0
		1-17 years old	87	9.0
		18-44 years old	284	29.2
		45-64 years old	266	27.4
		65-84 years old	187	19.2
		≥85 years old	11	1.1
		Unknown	1	0.1
	Gender	Female	498	51.2
		Male	474	48.8
Prospective Frozen	Age Group	<1 year	27	4.9
		1-17 years old	41	7.4
		18-44 years old	133	24.1
		45-64 years old	174	31.5
		65-84 years old	156	28.3
		≥85 years old	20	3.6
		Unknown	1	0.2
	Gender	Female	271	49.1
		Male	280	50.7
		Not available	1	0.2
Combined	Age Group	<1 year	163	10.7
		1-17 years old	128	8.4
		18-44 years old	417	27.4
		45-64 years old	440	28.9
		65-84 years old	343	22.5
		>85 years old	31	2.0

Table 15: Demographic data for prospective evaluated specimens

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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	SampleCategory	TP/TP+FN	%	95% CI*	TN/TN+FP	%	95% CI*
Bacteria
Escherichiacoli K1a	ProspectiveFresh	2/3	66.7	20.8-93.9	969 / 969	100.0	99.6-100.0
	ProspectiveFrozen	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

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QIAGEN

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

	Page 29 of 35

		Positive Percent Agreement		Negative Percent Agreement
Pathogen	SampleCategory	TP/TP+FN	%	95% CI*	TN/TN+FP	%	95% CI*
Bacteria
Haemophilusinfluenzae b	ProspectiveFresh	0 / 1	0.0	0.0-79.3	970 / 971	99.9	99.4-100.0
	ProspectiveFrozen	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
Listeriamonocytogenes c	ProspectiveFresh	1 / 1	100.0	20.7-100.0	971 / 971	100.0	99.6-100.0
	ProspectiveFrozen	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
Neisseriameningitidis(encapsulated)d	ProspectiveFresh	1 / 1	100.0	20.7-100.0	971 / 971	100.0	99.6-100.0
	ProspectiveFrozen	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
Streptococcusagalactiae	ProspectiveFresh	2 / 2	100.0	34.2-100.0	970 / 970	100.0	99.6-100.0
	ProspectiveFrozen	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
Streptococcuspneumoniae e	ProspectiveFresh	1 / 1	100.0	20.7-100.0	845 / 848	99.6	99.0-99.9
	ProspectiveFrozen	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
Streptococcuspyogenes	ProspectiveFresh	0 / 0	N/A	N/A	778 / 778	100.0	99.5-100.0
	ProspectiveFrozen	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	ProspectiveFresh	18 / 20	90.0	69.9-97.2	951 / 952	99.9	99.4-100.0
	ProspectiveFrozen	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
Cryptococcusgattii /Cryptococcusneoformans(notdifferentiated)g	ProspectiveFresh	2 / 5	40.0	11.8-76.9	965 / 967	99.8	99.2-99.9
	ProspectiveFrozen	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

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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	<1 year	11	26.8
		1-17 years old	9	22.0
		18-44 years old	12	29.3
		45-64 years old	5	12.2
		65-84 years old	4	9.8
Archived	Gender	Female	19	46.3
Archived	Gender	Male	22	53.7

The QIAstat-Dx ME Panel retrospective archived specimen data in positive percent agreement and negative percent against the comparator methods are presented by analyte in Table 18.

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	Positive Percent Agreement			Negative Percent Agreement
Pathogen	TP/TP+FN	%	95% CI	TN/TN+FP	%	95% CI
Overall Panel	41 / 41	100.0	91.4-100.0	314 / 314	100.0	98.8-100.0
	Bacteria
Escherichia coli Kl	2 / 2	100.0	34.2-100.0	39 / 39	100.0	91.0-100.0
Haemophilusinfluenzae	6 / 6	100.0	61.0-100.0	35 / 35	100.0	90.1-100.0
Listeriamonocytogenes	0 / 0	N/A	N/A	41 / 41	100.0	91.4-100.0
Neisseria meningitidis(encapsulated)	3 / 3	100.0	43.9-100.0	38 / 38	100.0	90.8-100.0
Streptococcusagalactiae	9 / 9	100.0	70.1-100.0	32 / 32	100.0	89.3-100.0
Streptococcuspneumoniae	4 / 4	100.0	51.0-100.0	18 / 18	100.0	82.4-100.0
Streptococcuspyogenes	0 / 0	N/A	N/A	23 / 23	100.0	85.7-100.0
Virus
Enterovirus (EV)	9 / 9	100.0	70.1-100.0	32 / 32	100.0	89.3-100.0
Fungi / Yeast
Cryptococcus gattii /Cryptococcusneoformans (notdifferentiated)	8 / 8	100.0	67.6-100.0	33 / 33	100.0	89.6-100.0

Table 18: Archived Clinical Performance Agreement (PPA) between QlAstat-Dx ME Panel and Comparator Method by Target*

*Please note that due to volume constraints, not all targets have the same number of specimens tested.

For each analyte where archived samples were available, a PPA of 100% was achieved.

Co-Infections

No specimens with multiple detections were identified by the QIAstat-Dx ME Panel during the performance evaluation.

Clinical sensitivity and specificity determined against culture

The performance measure of sensitivity and specificity was calculated only for bacterial and fungi / yeast analytes for which the gold-standard CSF culture result was available in the standard of care for the specimen. This data was used in additional performance calculations outlined in Table 19 and Table 20.

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		Sensitivity(compared to culture)			Specificity(compared to culture)
Pathogen	SampleCategory	TP/TP+FN	%	95% CI	TN/TN+FP	%	95% CI
Bacteria
Escherichiacoli Kl	Archived	1/1	100.0	20.7-100.0	10/11	90.9	62.3-98.4
	ProspectiveFresh	1/2	50.0	9.5-90.5	760/760	100.0	99.5-100.0
	ProspectiveFrozen	0/0	N/A	N/A	343/343	100.0	98.9-100.0
Haemophilusinfluenzae	Archived	1/1	100.0	20.7-100.0	10/11	90.9	62.3-98.4
	ProspectiveFresh	0/0	N/A	N/A	761/762	99.9	99.3-100.0
	ProspectiveFrozen	3/3	100.0	43.9-100.0	339/340	99.7	98.4-99.9
Listeriamonocytogenes	Archived	0/0	N/A	N/A	12/12	100.0	75.8-100.0
	ProspectiveFresh	1/1	100.0	20.7-100.0	761/761	100.0	99.5-100.0
	ProspectiveFrozen	2/3	66.7	20.8-93.9	340/340	100.0	98.9-100.0
Neisseriameningitidis(encapsulated)	Archived	2/2	100.0	34.2-100.0	9/10	90.0	59.6-98.2
	ProspectiveFresh	0/0	N/A	N/A	761/762	99.9	99.3-100.0
	ProspectiveFrozen	0/0	N/A	N/A	342/343	99.7	98.4-99.9
Streptococcusagalactiae	Archived	0/0	N/A	N/A	12/12	100.0	75.8-100.0
	ProspectiveFresh	1/1	100.0	20.7-100.0	760/761	99.9	99.3-100.0
	ProspectiveFrozen	1/1	100.0	20.7-100.0	342/342	100.0	98.9-100.0
Streptococcuspneumoniae	Archived	0/0	N/A	N/A	11/12	91.7	64.6-98.5
	ProspectiveFresh	0/0	N/A	N/A	757/762	99.3	98.5-99.7
	ProspectiveFrozen	3/3	100.0	43.9-100.0	339/340	99.7	98.4-99.9
Streptococcuspyogenes	Archived	0/0	N/A	N/A	12/12	100.0	75.8-100.0
	ProspectiveFresh	0/0	N/A	N/A	761/762	99.9	99.3-100.0
	ProspectiveFrozen	0/0	N/A	N/A	343/343	100.0	98.9-100.0

Table 19: Bacterial Culture comparison for diagnostic sensitivity and specificity by sample category

Cryptococcus was also calculated in comparison to fungal culture performed as a standard of care diagnostic at clinical testing sites. For data that were available, QIAstat-Dx ME Panel performance is summarized in Table 20.

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		Positive Percent Agreement			Negative Percent Agreement
Pathogen	Sample Category	TP/TP+FN	%	95% CI	TN/TN+FP	%	95% CI
Fungi / Yeast
Cryptococcusgattii	Archived	2 / 2	100.0	34.2-100.0	1 / 1	100.0	20.7-100.0
	Prospective	1 / 1	100.0	20.7-100.0	129 / 131	98.5	94.6-99.6
Cryptococcusneoformans(notdifferentiated)	Fresh
	ProspectiveFrozen	0 / 0	N/A	N/A	24 / 24	100.0	86.2-100.0

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.

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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	ConcentrationLevel	Frequencyof PositiveResults	Proportion(%) ofPositiveResults	Lower95%ConfidenceLimit	Upper95%ConfidenceLimit
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
	Streptococcusagalactiae	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
	Streptococcuspneumoniae	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 /Cryptococcusneoformans (notdifferentiated)	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

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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.

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.