Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a standard real-time RT-PCR assay and instrument for detecting SARS-CoV-2 RNA. There is no mention of AI, ML, or any related technologies in the device description, intended use, or performance studies. The analysis is based on standard molecular diagnostic techniques.

Therapeutic

No.

The device is an in vitro diagnostic (IVD) assay used to detect SARS-CoV-2. It is an "aid in the diagnosis of SARS-CoV-2 infection" and its results are "meant to be used in conjunction with other clinical, epidemiologic, and laboratory data". It does not describe any function that directly treats or prevents a disease, which are characteristics of a therapeutic device.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states, "The Simplexa™ COVID-19 Direct assay is an aid in the diagnosis of SARS-CoV-2 infection." This directly indicates its role as a diagnostic device.

SaMD (Software as a Medical Device)

No

The device description explicitly states that the system consists of the Simplexa COVID-19 Direct reaction mix, the LIAISON MDX (with LIAISON MDX Studio Software), the Direct Amplification Disc and associated accessories. This includes physical components beyond just software.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the assay is "intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of nucleic acid from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)". The term "in vitro" means "in glass" or "in the lab," referring to tests performed outside of a living organism.
Purpose: The assay is designed to detect the presence of SARS-CoV-2 RNA in patient samples (nasopharyngeal and nasal swabs) to aid in the diagnosis of COVID-19. This is a classic function of an in vitro diagnostic device.
Sample Type: It uses biological samples (swabs) collected from the patient, which are then analyzed in a laboratory setting.
Device Description: The description details a laboratory-based system (real-time RT-PCR) that analyzes the nucleic acid from the sample.

All these points align with the definition of an In Vitro Diagnostic device, which is a medical device intended for use in vitro for the examination of specimens derived from the human body solely or principally for the purpose of providing information concerning a physiological or pathological state, or concerning a congenital abnormality, or to monitor therapeutic measures.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The DiaSorin Molecular Simplexa™ COVID-19 Direct is real-time RT-PCR assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of nucleic acid from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal swabs (NPS) and nasal swabs (NS) from symptomatic individuals suspected of COVID 19 by their healthcare provider. The Simplexa™ COVID-19 Direct assay is an aid in the diagnosis of SARS-CoV-2 infection.

Positive results are indicative of the presence of SARS-CoV-2 RNA. Clinical correlation with patient history and other diagnostic information is necessary to determine patient infection status. Positive results do not rule out co-infection with other pathogens. Negative results do not preclude SARS-CoV-2 infection and should not be used as tor patient management decisions.

Results are meant to be used in conjunction with other clinical, epidemiologic, and laboratory data, in accordance with the guidelines provided by the relevant public health authorities.

Product codes

OOX, OOI

Device Description

The Simplexa COVID-19 Direct is a real-time RT-PCR (rRT-PCR) system that enables the direct amplification and detection of SARS-CoV-2 (COVID-19) RNA from nasopharyngeal swab or nasal swab that has not undergone nucleic acid extraction. The system consists of the Simplexa COVID-19 Direct reaction mix, the LIAISON MDX (with LIAISON MDX Studio Software), the Direct Amplification Disc and associated accessories. The assay uses forward and reverse primers and associated fluorescent probe(s) included in the reaction mix to amplify SARS-CoV-2 cDNA reverse transcribed from RNA. The primers and probe sets are designed to detect SARS-CoV-2 ORF 1ab and S gene from the viral RNA in nasopharyngeal swab or nasal swab. An RNA internal control, with associated primers and a fluorescent probe, is included in the reaction mix to detect RT-PCR failure and/or inhibition.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasopharyngeal swabs (NPS); nasal swabs (NS)

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

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

Not Found

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

The clinical performance of the Simplexa™ COVID-19 Direct was established in multi-site clinical evaluation. A total of 1,150 prospective fresh and/or prospective frozen Nasopharyngeal (NPS) and/or Nasal Swabs (NS) samples were collected from four (4) geographically diverse collection sites, one of which was outside the United States (OUS). A total of 409 NPS samples and a total of 741 nasal swabs were collected. One hundred fourteen (114) samples were excluded from the analysis due to insufficient evidence to include certain media types, 24 were excluded due to invalid results, and one (1) specimen was excluded due to an indeterminate result from the composite reference method. The Simplexa™ COVID-19 Direct Clinical Agreement testing was performed at three (3) external clinical sites and one (1) internal site from October 2020 to April 2021. Three COVID-19 EUA approved NAAT assays were used to establish a comparator reference method (CRM) to evaluate the performance of the candidate device. Two out of three positive results determined "Detected" CRM and two out of three negative results determined "Not Detected" CRM.

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

CLINICAL AGREEMENT
Study Type: Clinical evaluation
Sample Size: 1011 total (1,150 collected, 114 excluded due to insufficient evidence, 24 excluded due to invalid results, 1 excluded due to indeterminate CRM result)
Results:

Total Specimen Clinical Summary:
- PPA: 98.2% (108/110) with 95% CI: 93.6% to 99.5%
- NPA: 99.6% (897/901) with 95% CI: 98.9% to 99.8%
NPS Clinical Summary:
- PPA: 98.4% (60/61) with 95% CI: 91.3% to 99.7%
- NPA: 99.6% (237/238) with 95% CI: 97.7% to 99.9%
NS Collection Clinical Summary:
- PPA: 98.0% (48/49) with 95% CI: 89.3% to 99.6%
- NPA: 99.5% (660/663) with 95% CI: 98.7% to 99.8%

REPRODUCIBILITY
Study Type: Reproducibility study
Sample Size: 90 replicates per panel member (total of 4 panel members)
Results:

NPS_WA-1_LP:
- S gene: Agreement with Expected Results 94.4% (85/90), 95% CI: 88%-98%
- ORF1ab: Agreement with Expected Results 95.6% (86/90), 95% CI: 89%-98%
- Total (algorithm based): Agreement with Expected Results 98.9% (89/90), 95% CI: 94%-100%
NPS_WA-1_MP:
- S gene: Agreement with Expected Results 95.6% (86/90), 95% CI: 89%-98%
- ORF1ab: Agreement with Expected Results 100.0% (90/90), 95% CI: 96%-100%
- Total (algorithm based): Agreement with Expected Results 100.0% (90/90), 95% CI: 96%-100%
Negative (UTM):
- S gene: Agreement with Expected Results 100.0% (90/90), 95% CI: 96%-100%
- ORF1ab: Agreement with Expected Results 100.0% (90/90), 95% CI: 96%-100%
- Total (algorithm based): Agreement with Expected Results 100.0% (90/90), 95% CI: 96%-100%
Positive Control (PC):
- S gene: Agreement with Expected Results 100.0% (90/90), 95% CI: 96%-100%
- ORF1ab: Agreement with Expected Results 100.0% (90/90), 95% CI: 96%-100%
- Total (algorithm based): Agreement with Expected Results 100.0% (90/90), 95% CI: 96%-100%

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION
Study Type: LoD determination
Results:

Inactivated Viral Particles:
- NPS LoD: 500 copies/mL (100% detection for SARS-CoV-2 (40/40), 90% for S gene (36/40), 100% for ORF1ab (40/40))
- NS LoD: 242 copies/mL (100% detection for SARS-CoV-2 (20/20), 80% for S gene (16/20), 80% for ORF1ab (16/20))
WHO International Standard:
- NPS LoD: 500 IU/mL (97.5% detection for SARS-CoV-2 (39/40), 95% for S gene (38/40), 85% for ORF1ab (34/40))
FDA SARS-CoV-2 Reference Panel:
- Product LoD: 6 x 10^3 NDU/mL (NPS)

REACTIVITY/INCLUSIVITY
Study Type: In silico and wet testing
Results:

In Silico:
- Nov. 1, 2021 to Jan. 31, 2022: 100% identity to SARS-CoV-2 gene design (2,170,584 sequences analyzed)
- Feb. 1, 2022 to Apr. 30, 2022: 100% identity to SARS-CoV-2 gene design (377,668 sequences analyzed)
- May 1, 2022 to July 31, 2022: 100% identity to SARS-CoV-2 gene design (211,224 sequences analyzed)
Wet Testing:
- All tested SARS-CoV-2 strains (Hong Kong, England, South Africa, Japan, hCoV19) detected at 100% (3/3 replicates) at 1000 copies/mL concentration.

CROSS-REACTIVITY
Study Type: In silico and wet testing
Results: No cross-reactivity observed with 47 different viruses, bacteria, and fungi, and pooled human nasal fluid. All samples showed 0.0% detection for S gene and ORF1ab, and 100.0% detection for the Internal Control (IC).

POTENTIAL INTERFERING SUBSTANCES
Study Type: Interference testing
Results:

Most substances showed 100% detection for both COVID-19 Qualitative Results and IC Qualitative Results.
Saliva showed 83.3% detection (5/6) for both COVID-19 and IC Qualitative Results at 10% (v/v), but 100% detection at 5% (v/v). Interference from saliva was observed at a concentration above 5%.
Zanamivir showed 100% detection (6/6) for COVID-19 Qualitative Results and 83.3% detection (5/6) for IC Qualitative Results.

INTERFERENCE BY OTHER MICROORGANISMS
Study Type: Microbial interference
Results: No inhibition by other organisms was observed at the tested concentrations for 47 different organisms and pooled human nasal fluid. All tested organisms showed 100.0% detection for COVID-19 Qualitative Results at 2x LoD of inactivated COVID-19 viral particles, except for Mumps (95.0% detection, 19/20). Lactobacillus plantarum 17-5 showed interference when tested above 5 x 10^5 CFU/mL.

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

Clinical Agreement:

PPA (Percent Positive Agreement): 98.2% (108/110) with 95% CI: 93.6% to 99.5% (Total Specimen)
NPA (Negative Percent Agreement): 99.6% (897/901) with 95% CI: 98.9% to 99.8% (Total Specimen)

Analytical Sensitivity/Limit of Detection (LoD):

NPS LoD: 500 copies/mL (100% detection)
NS LoD: 242 copies/mL (100% detection)
NPS LoD using WHO International Standard: 500 IU/mL (97.5% detection)
FDA SARS-CoV-2 Reference Panel LoD: 6 x 10^3 NDU/mL

Predicate Device(s)

BioFire COVID-19 Test 2 (K211079)

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3981 Device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test.

(a)
Identification. A device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test is an in vitro diagnostic device intended for the detection and identification of SARS-CoV-2 and other microbial agents when in a multi-target test in human clinical respiratory specimens from patients suspected of respiratory infection who are at risk for exposure or who may have been exposed to these agents. The device is intended to aid in the diagnosis of respiratory infection in conjunction with other clinical, epidemiologic, and laboratory data or other risk factors.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The intended use in the labeling required under § 809.10 of this chapter must include a description of the following: Analytes and targets the device detects and identifies, the specimen types tested, the results provided to the user, the clinical indications for which the test is to be used, the specific intended population(s), the intended use locations including testing location(s) where the device is to be used (if applicable), and other conditions of use as appropriate.
(2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device; alternatively, the sample collection device must be cleared in a premarket submission as a part of this device.
(3) The labeling required under § 809.10(b) of this chapter must include:
(i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens;
(ii) Detailed descriptions of the performance characteristics of the device for each specimen type claimed in the intended use based on analytical studies including the following, as applicable: Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, precision, reproducibility, and clinical studies;
(iii) Detailed descriptions of the test procedure(s), the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing;
(iv) A warning statement that viral culture should not be attempted in cases of positive results for SARS-CoV-2 and/or any similar microbial agents unless a facility with an appropriate level of laboratory biosafety (
e.g., BSL 3 and BSL 3+, etc.) is available to receive and culture specimens; and(v) A prominent statement that device performance has not been established for specimens collected from individuals not identified in the intended use population (
e.g., when applicable, that device performance has not been established in individuals without signs or symptoms of respiratory infection).(vi) Limiting statements that indicate that:
(A) A negative test result does not preclude the possibility of infection;
(B) The test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician;
(C) There is a risk of incorrect results due to the presence of nucleic acid sequence variants in the targeted pathogens;
(D) That positive and negative predictive values are highly dependent on prevalence;
(E) Accurate results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe proper procedures in any one of these steps can lead to incorrect results; and
(F) When applicable (
e.g., recommended by the Centers for Disease Control and Prevention, by current well-accepted clinical guidelines, or by published peer-reviewed literature), that the clinical performance may be affected by testing a specific clinical subpopulation or for a specific claimed specimen type.(4) Design verification and validation must include:
(i) Detailed documentation, including performance results, from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, as appropriate, additional characterized clinical samples. The clinical study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained using a comparator that FDA has determined is appropriate. Detailed documentation must include the clinical study protocol (including a predefined statistical analysis plan), study report, testing results, and results of all statistical analyses.
(ii) Risk analysis and documentation demonstrating how risk control measures are implemented to address device system hazards, such as Failure Modes Effects Analysis and/or Hazard Analysis. This documentation must include a detailed description of a protocol (including all procedures and methods) for the continuous monitoring, identification, and handling of genetic mutations and/or novel respiratory pathogen isolates or strains (
e.g., regular review of published literature and periodic in silico analysis of target sequences to detect possible mismatches). All results of this protocol, including any findings, must be documented and must include any additional data analysis that is requested by FDA in response to any performance concerns identified under this section or identified by FDA during routine evaluation. Additionally, if requested by FDA, these evaluations must be submitted to FDA for FDA review within 48 hours of the request. Results that are reasonably interpreted to support the conclusion that novel respiratory pathogen strains or isolates impact the stated expected performance of the device must be sent to FDA immediately.(iii) A detailed description of the identity, phylogenetic relationship, and other recognized characterization of the respiratory pathogen(s) that the device is designed to detect. In addition, detailed documentation describing how to interpret the device results and other measures that might be needed for a laboratory diagnosis of respiratory infection.
(iv) A detailed device description, including device components, ancillary reagents required but not provided, and a detailed explanation of the methodology, including molecular target(s) for each analyte, design of target detection reagents, rationale for target selection, limiting factors of the device (
e.g., saturation level of hybridization and maximum amplification and detection cycle number, etc.), internal and external controls, and computational path from collected raw data to reported result (e.g., how collected raw signals are converted into a reported signal and result), as applicable.(v) A detailed description of device software, including software applications and hardware-based devices that incorporate software. The detailed description must include documentation of verification, validation, and hazard analysis and risk assessment activities, including an assessment of the impact of threats and vulnerabilities on device functionality and end users/patients as part of cybersecurity review.
(vi) For devices intended for the detection and identification of microbial agents for which an FDA recommended reference panel is available, design verification and validation must include the performance results of an analytical study testing the FDA recommended reference panel of characterized samples. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(vii) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens, the design verification and validation must include a detailed description of the identity, phylogenetic relationship, or other recognized characterization of the Influenza A and B viruses that the device is designed to detect, a description of how the device results might be used in a diagnostic algorithm and other measures that might be needed for a laboratory identification of Influenza A or B virus and of specific Influenza A virus subtypes, and a description of the clinical and epidemiological parameters that are relevant to a patient case diagnosis of Influenza A or B and of specific Influenza A virus subtypes. An evaluation of the device compared to a currently appropriate and FDA accepted comparator method. Detailed documentation must be kept of that study and its results, including the study protocol, study report for the proposed intended use, testing results, and results of all statistical analyses.
(5) When applicable, performance results of the analytical study testing the FDA recommended reference panel described in paragraph (b)(4)(vi) of this section must be included in the device's labeling under § 809.10(b) of this chapter.
(6) For devices with an intended use that includes detection of Influenza A and Influenza B viruses and/or detection and differentiation between the Influenza A virus subtypes in human clinical specimens in addition to detection of SARS-CoV-2 and similar microbial agents, the required labeling under § 809.10(b) of this chapter must include the following:
(i) Where applicable, a limiting statement that performance characteristics for Influenza A were established when Influenza A/H3 and A/H1-2009 (or other pertinent Influenza A subtypes) were the predominant Influenza A viruses in circulation.
(ii) Where applicable, a warning statement that reads if infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to State or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
(iii) Where the device results interpretation involves combining the outputs of several targets to get the final results, such as a device that both detects Influenza A and differentiates all known Influenza A subtypes that are currently circulating, the device's labeling must include a clear interpretation instruction for all valid and invalid output combinations, and recommendations for any required followup actions or retesting in the case of an unusual or unexpected device result.
(iv) A limiting statement that if a specimen yields a positive result for Influenza A, but produces negative test results for all specific influenza A subtypes intended to be differentiated (
i.e., H1-2009 and H3), this result requires notification of appropriate local, State, or Federal public health authorities to determine necessary measures for verification and to further determine whether the specimen represents a novel strain of Influenza A.(7) If one of the actions listed at section 564(b)(1)(A) through (D) of the Federal Food, Drug, and Cosmetic Act occurs with respect to an influenza viral strain, or if the Secretary of Health and Human Services determines, under section 319(a) of the Public Health Service Act, that a disease or disorder presents a public health emergency, or that a public health emergency otherwise exists, with respect to an influenza viral strain:
(i) Within 30 days from the date that FDA notifies manufacturers that characterized viral samples are available for test evaluation, the manufacturer must have testing performed on the device with those influenza viral samples in accordance with a standardized protocol considered and determined by FDA to be acceptable and appropriate.
(ii) Within 60 days from the date that FDA notifies manufacturers that characterized influenza viral samples are available for test evaluation and continuing until 3 years from that date, the results of the influenza emergency analytical reactivity testing, including the detailed information for the virus tested as described in the certificate of authentication, must be included as part of the device's labeling in a tabular format, either by:
(A) Placing the results directly in the device's labeling required under § 809.10(b) of this chapter that accompanies the device in a separate section of the labeling where analytical reactivity testing data can be found, but separate from the annual analytical reactivity testing results; or
(B) In a section of the device's label or in other labeling that accompanies the device, prominently providing a hyperlink to the manufacturer's public website where the analytical reactivity testing data can be found. The manufacturer's website, as well as the primary part of the manufacturer's website that discusses the device, must provide a prominently placed hyperlink to the website containing this information and must allow unrestricted viewing access.

Summary

0

September 13, 2022

Image /page/0/Picture/1 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

DiaSorin Molecular LLC Tara Viviani Director, Regulatory Affairs 11331 Valley View Street Cypress, California 90630

Re: K212147

Trade/Device Name: Simplexa COVID-19 Direct Regulation Number: 21 CFR 866.3981 Regulation Name: Device to detect and identify nucleic acid targets in respiratory specimens from microbial agents that cause the SARS-CoV-2 respiratory infection and other microbial agents when in a multi-target test Regulatory Class: Class II Product Code: OOX Dated: August 12, 2022 Received: August 15, 2022

Dear Tara Viviani:

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 (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 located 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.

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

1

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 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-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 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 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).

Sincerely,

Himani Bisht. Ph.D. Assistant Director Viral Respiratory and HPV Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K212147

Device Name Simplexa™ COVID-19 Direct

Indications for Use (Describe)

The DiaSorin Molecular Simplexa™ COVID-19 Direct is real-time RT-PCR assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of nucleic acid from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal swabs (NPS) and nasal swabs (NS) from symptomatic individuals suspected of COVID 19 by their healthcare provider. The Simplexa™ COVID-19 Direct assay is an aid in the diagnosis of SARS-CoV-2 infection.

Positive results are indicative of the presence of SARS-CoV-2 RNA. Clinical correlation with patient history and other diagnostic information is necessary to determine patient infection status. Positive results do not rule out co-infection with other pathogens. Negative results do not preclude SARS-CoV-2 infection and should not be used as tor patient management decisions.

Results are meant to be used in conjunction with other clinical, epidemiologic, and laboratory data, in accordance with the guidelines provided by the relevant public health authorities.

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.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

3

Image /page/3/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the DNA graphic are the words "DiaSorin" in dark blue on the top line and "Molecular" in green on the bottom line.

| Applicant | DiaSorin Molecular LLC.
11331 Valley View Street
Cypress, California 90630
USA |
|--------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Establishment Registration No. | 2023365 |
| Contact Person | Tara Viviani
Senior Director, Molecular Regulatory Affairs
Tel. 562.240.6271
Tara. Viviani@DiaSorin.com |
| Summary Date | September 13, 2022 |
| Proprietary Name | Simplexa™ COVID-19 Direct |
| US Product Codes/Names and
Regulation Numbers | QQX- Device to detect and identify nucleic acid targets in respiratory
specimens from microbial agents that cause the SARS-CoV-2
respiratory infection and other microbial agents when in a multi-
target test (21 CFR § 866.3981)
OOI - Instrumentation for clinical multiplex test systems (21 CFR §
862.2570) |
| Classification | Class II |
| Predicate Devices | BioFire COVID-19 Test 2 (K211079) |

Intended Use

Simplexa™ COVID-19 Direct REF MOL4150

The DiaSorin Molecular Simplexa™ COVID-19 Direct is a real-time RT-PCR assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of nucleic acid from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal swabs (NPS) and nasal swab specimens from symptomatic individuals suspected of COVID-19 by their healthcare provider. The Simplexa™ COVID-19 Direct assay is an aid in the diagnosis of SARS-CoV-2 infection.

Positive results are indicative of the presence of SARS-CoV-2 RNA. Clinical correlation with patient history and other diagnostic information is necessary to determine patient infection status. Positive results do not rule out co-infection with other pathogens. Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for patient management decisions.

Results are meant to be used in conjunction with other clinical, epidemiologic, and laboratory data, in accordance with the guidelines provided by the relevant public health authorities.

Device Description

The Simplexa COVID-19 Direct is a real-time RT-PCR (rRT-PCR) system that enables the direct amplification and detection of SARS-CoV-2 (COVID-19) RNA from nasopharyngeal swab or nasal swab that has not undergone nucleic acid extraction. The system consists of the Simplexa COVID-19 Direct reaction mix, the LIAISON MDX (with LIAISON MDX Studio Software), the Direct Amplification Disc and

4

Image /page/4/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a stylized DNA helix in shades of green and blue on the left, followed by the text "DiaSorin" in dark blue on the top line and "Molecular" in green on the bottom line. The logo is clean and modern, suggesting a focus on biotechnology and molecular diagnostics.

associated accessories. The assay uses forward and reverse primers and associated fluorescent probe(s) included in the reaction mix to amplify SARS-CoV-2 cDNA reverse transcribed from RNA. The primers and probe sets are designed to detect SARS-CoV-2 ORF 1ab and S gene from the viral RNA in nasopharyngeal swab or nasal swab. An RNA internal control, with associated primers and a fluorescent probe, is included in the reaction mix to detect RT-PCR failure and/or inhibition.

Simplexa™ COVID-19 Direct REF MOL4150

| Component Name | REF | EC Symbol
on Label | Abbreviated
Name | Cap
Color | Number
of Vials | Reactions
per
Vial/Kit | Volume
per Vial |
|-------------------------------------------|---------|-----------------------|---------------------|--------------|--------------------|------------------------------|--------------------|
| Simplexa™ COVID-19
Direct Reaction Mix | MOL4151 | REAG | Co19 | Brown | 24 | 1/24 | 50 $ μ $ L |

Simplexa™ COVID-19 Direct Components and Descriptions

Kit Component	Contents
Simplexa™
COVID-19 Direct
Reaction Mix (RM)	DNA polymerase, reverse transcriptase, RNase inhibitor, buffer, dNTPs, encapsulated RNA template (Internal Control), fluorescent probes and corresponding forward and reverse primers specific for detection of SARS-CoV-2 RNA and for the RNA Internal Control.
	Target	Probe
Fluorophore
(Dye)	Excitation (nm)	Emission
(nm)	Targeted Gene
	S gene	FAM	445-505	507-533	S gene
	ORF1ab	JOE	503-543	547-573	ORF1ab
	Internal
Control RNA
(IC)	Q670	622-658	652-708	N/A
Simplexa™
COVID-19 Direct
Barcode	Assay specific parameters and lot information

Materials Supplied Separately

Direct Amplification Disc Kit (REF MOL1455) - Direct Amplification Discs for use on the LIAISON® MDX

Materials Required But Not Supplied

LIAISON® MDX with LIAISON® MDX Studio Software version 1.1 or higher.

Simplexa™ COVID-19 Positive Control Gen II Pack (REF MOL4165).

50 µL fixed volume pipette (VWR Signature™ Fixed Volume Ergonomic High-Performance Pipettor Model VWR FE50 or equivalent).

Sterile, nuclease-free disposable pipette tips with filters (Extra Long tips ≥ 91 mm are recommended for pipetting directly from primary collection tubes).

Freezer (manual defrost) at -10 to -30 °C (for kit component and/or specimen frozen storage).

Refrigerator at 2 to 8 °C (for specimens).

Disposable, powder-free gloves.

Vortex for mixing patient samples.

Centrifuge for collecting contents to bottom of tubes.

5

Image /page/5/Picture/0 description: The image contains the logo for DiaSorin Molecular. The logo consists of a stylized DNA double helix on the left, rendered in shades of green and blue. To the right of the helix, the word "DiaSorin" is written in a dark blue, serif font. Below "DiaSorin", the word "Molecular" is written in a lighter green color, also in a serif font.

510(k) Summary Simplexa™ COVID-19 Direct MOL4150 September 12, 2022 Page 3 of 13

Comparison to Predicate Device

| Comparison to
Predicate Device | Predicate Device:
BioFire COVID-19 Test 2 (K211079) | Candidate Device:
Simplexa COVID-19 Direct
Simplex COVID-19 Positive Control Gen II Pack |
|-------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Product Code | QQX | Same |
| Regulation
Number | 21 CFR 866.3981 | Same |
| Organism
Detected | SARS-CoV-2 | Same |
| Measurand | RNA from SARS-CoV-2 | Same |
| Intended Use Kit | The BioFire COVID-19 Test 2 is a
qualitative nested multiplexed RT-PCR in
vitro diagnostic test intended for use with
the BioFire FilmArray 2.0 and BioFire
FilmArray Torch Systems. The BioFire
COVID-19 Test 2 detects nucleic acids from
severe acute respiratory syndrome
coronavirus 2 (SARS-CoV-2) in
nasopharyngeal swabs (NPS) from
symptomatic individuals suspected of
COVID-19 by their healthcare provider.
Results are for the identification of SARS-
Co V-2 RNA. The SARS-Co V-2 RNA is
generally detectable in NPS specimens
during the acute phase of infection. Positive
results are indicative of the presence of
SARS-CoV-2 RNA; clinical correlation with
patient history and other diagnostic
information is necessary to determine
patient infection status. Positive results do
not rule out co-infection with other
pathogens. Results are meant to be used in
conjunction with other clinical,
epidemiologic, and laboratory data, in
accordance with the guidelines provided by
the relevant public health authorities. The
BioFire COVID-19 Test 2 is intended for use
by trained medical and laboratory
professionals in a laboratory setting or
under the supervision of a trained laboratory
professional. | Simplexa™ COVID-19 Direct REF MOL4150

The DiaSorin Molecular Simplexa™ COVID-19
Direct is a real-time RT-PCR assay intended for
use on the LIAISON® MDX instrument for the in
vitro qualitative detection of nucleic acid from
severe acute respiratory syndrome coronavirus
2 (SARS-CoV-2) in nasopharyngeal swabs
(NPS) and nasal swabs (NS) from symptomatic
individuals suspected of COVID-19 by their
healthcare provider. The Simplexa™ COVID-19
Direct assay is an aid in the diagnosis of SARS-
CoV-2 infection.

Positive results are indicative of the presence of
SARS-CoV-2 RNA. Clinical correlation with
patient history and other diagnostic information
is necessary to determine patient infection
status. Positive results do not rule out co-
infection with other pathogens. Negative results
do not preclude SARS-CoV-2 infection and
should not be used as the sole basis for patient
management decisions.

CLINICAL AGREEMENT

The clinical performance of the Simplexa™ COVID-19 Direct was established in multi-site clinical evaluation. A total of 1,150 prospective fresh and/or prospective frozen Nasopharyngeal (NPS) and/or

6

Image /page/6/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a green and blue DNA helix graphic on the left, with the text "DiaSorin" in dark blue to the right of the helix. Below "DiaSorin" is the word "Molecular" in green.

510(k) Summarv Simplexa™ COVID-19 Direct MOL4150 September 12, 2022 Page 4 of 13

Nasal Swabs (NS) samples were collected from four (4) geographically diverse collection sites, one of which was outside the United States (OUS). A total of 409 NPS samples and a total of 741 nasal swabs were collected. One hundred fourteen (114) samples were excluded from the analysis due to insufficient evidence to include certain media types, 24 were excluded due to invalid results, and one (1) specimen was excluded due to an indeterminate result from the composite reference method. The Simplexa™ COVID-19 Direct Clinical Agreement testing was performed at three (3) external clinical sites and one (1) internal site from October 2020 to April 2021. Three COVID-19 EUA approved NAAT assays were used to establish a comparator reference method (CRM) to evaluate the performance of the candidate device. Two out of three positive results determined "Detected" CRM and two out of three negative results determined "Not Detected" CRM. The clinical performance of Simplexa™ COVID-19 Direct is summarized in the Tables 1-3 below.

Table 1: Total Specimen Clinical Summary: Simplexa™ COVID-19 Direct vs. EUA NAAT CRM

| Simplexa™ COVID-19 Direct

Prospective	CRM
Upper Respiratory	Detected	Not Detected	Total
Detected	108	4	112
Not Detected	2	897	899
Total	110	901	1011
PPA:	98.2% (108/110)
95% CI: 93.6% to 99.5%
NPA:	99.6% (897/901)
95% CI: 98.9% to 99.8%

Table 2: NPS Clinical Summary: Simplexa™ COVID-19 Direct vs. EUA NAAT CRM

| Simplexa™ COVID-19 Direct

Prospective NPS	CRM
	Detected	Not Detected	Total
Detected	60	1a	61
Not Detected	1	237	238
Total	61	238	299
PPA:	98.4% (60/61)
95% CI: 91.3% to 99.7%
NPA:	99.6% (237/238)
95% CI: 97.7% to 99.9%

ª One sample positive by Simplexa was detected one of the NAATs included in the CRM.

CRM = Composite Reference Method, PPA = Percent Positive Agreement, NPA = Negative Percent Agreement

7

Image /page/7/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a stylized DNA double helix on the left, with the words "DiaSorin" in dark blue on the right. Below "DiaSorin" is the word "Molecular" in green. The DNA helix is colored with a gradient from green to blue.

Table 3: NS Collection: Simplexa™ COVID-19 Direct vs. EUA NAAT CRM

| Simplexa™ COVID-19 Direct
Prospective

NS	CRM
	Detected	Not Detected	Total
Detected	48	3a	51
Not Detected	1	660	661
Total	49	663	712
PPA:	98.0% (48/49)
95% CI: 89.3% to 99.6%
NPA:	99.5% (660/663)
95% CI: 98.7% to 99.8%

a One sample positive by Simplexa was positive by one of the NAATs in the CRM.

CRM = Composite Reference Method, PPA = Percent Positive Agreement, NPA = Negative Percent Agreement

REPRODUCIBILITY

The Simplexa™ COVID-19 Direct reproducibility study was performed at two (2) external clinical sites and one (1) internal site to evaluate repeatability, between operator, between site and total reproducibility of the assay. The panel consisted of a total of four (4) panel members including two (2) contrived panel samples, one (1) positive control and one (1) universal transport media (UTM) neqative sample. The contrived panel members were prepared at approximately one to two times (1x to 2x) the Limit of Detection (LoD) for a low positive (LP) and three to five times (3x to 5x) LoD for a moderate positive (MP). Both contrived panel members were built with SARS-CoV-2 strain 2019-nCoV/USA-WA1/2020 spiked into native negative nasopharyngeal swab matrix. Each panel member was tested in triplicate per run for two (2) runs per day for a total of five (5) non-consecutive testing days at three (3) testing sites for a total of ninety (90) replicates. Each site had two (2) operators who each assayed the entire panel once per day, for a total of two (2) sets of data per day. The quantitative summary of variance components for each panel member based on the algorithm is in Table 4 below.

		Table 4. Simplexa™ COVID-19 Direct Reproducibility

		Site 1
Sample		Agree-
ment with
Expected
Results	Avg.
Ct ±
SD
(%CV)	Agree-
ment with
Expected
Results	Avg.
Ct ±
SD
(%CV)	Agree-
ment with
Expected
Results	Avg.
Ct ±
SD
(%CV)	Agree-
ment with
Expected
Results	95% CI
NPS_WA-
1_LP	S gene	93.3%
(28/30)	31.6 ±
0.95
(3.0%)	90.0%
(27/30)	32.4 ±
1.14
(3.5%)	100.0%
(30/30)	31.6 ±
0.72
(2.3%)	94.4%
(85/90)	88% -
98%
	ORF1ab	90.0%
(27/30)	32.2 ±
0.97
(3.0%)	96.7%
(29/30)	32.7 ±
1.18
(3.6%)	100.0%
(30/30)	31.9 ±
1.09
(3.4%)	95.6%
(86/90)	89% -
98%
	Totala	100.0%
(30/30)	N/A	96.7%
(29/30)	N/A	100.0%
(30/30)	N/A	98.9%
(89/90)	94% -
100%
NPS_WA-
1_MP	S gene	100.0%
(30/30)	30.5 ±
0.80
(2.6%)	90.0%
(27/30)	30.8 ±
1.11
(3.6%)	96.7%
(29/30)	31.1 ±
1.14
(3.7%)	95.6%
(86/90)	89% -
98%

8

Image /page/8/Picture/0 description: The image shows the text 'K212147' in a bold, sans-serif font. The text is arranged horizontally, with the letter 'K' followed by the numbers '212147'. The text is black and the background is white.

Image /page/8/Picture/1 description: The image shows the logo for DiaSorin Molecular. The logo consists of a green DNA helix graphic on the left, followed by the text "DiaSorin" in dark blue. Below "DiaSorin" is the word "Molecular" in green. The logo is clean and professional, suggesting a company involved in molecular diagnostics or research.

Simplexa™ COVID-

Sample		Site 1	Site 2	Site 4	All Sites
		Agree-
ment with
Expected
Results	Avg.
Ct ±
SD
(%CV)	Agree-
ment with
Expected
Results	Avg.
Ct ±
SD
(%CV)	Agree-
ment with
Expected
Results	Avg.
Ct ±
SD
(%CV)	Agree-
ment with
Expected
Results	95% CI
	ORF1ab	100.0%
(30/30)	31.3 ±
0.87
(2.8%)	100.0%
(30/30)	31.6 ±
1.39
(4.4%)	100.0%
(30/30)	30.7 ±
0.98
(3.2%)	100.0%
(90/90)	96% -
100%
	Totala	100.0%
(30/30)	N/A	100.0%
(30/30)	N/A	100.0%
(30/30)	N/A	100.0%
(90/90)	96% -
100%
Negative
(UTM)	S gene	100.0%
(30/30)	0.0 ±
0.00
(N/A%)	100.0%
(30/30)	0.0 ±
0.00
(N/A%)	100.0%
(30/30)	0.0 ±
0.00
(N/A%)	100.0%
(90/90)	96% -
100%
	ORF1ab	100.0%
(30/30)	0.0 ±
0.00
(N/A%)	100.0%
(30/30)	0.0 ±
0.00
(N/A%)	100.0%
(30/30)	0.0 ±
0.00
(N/A%)	100.0%
(90/90)	96% -
100%
	Totala	100.0%
(30/30)	N/A	100.0%
(30/30)	N/A	100.0%
(30/30)	N/A	100.0%
(90/90)	96% -
100%
Positive
Control
(PC)	S gene	100.0%
(30/30)	25.8 ±
0.26
(1.0%)	100.0%
(30/30)	25.7 ±
0.18
(0.7%)	100.0%
(30/30)	25.7 ±
0.42
(1.6%)	100.0%
(90/90)	96% -
100%
	ORF1ab	100.0%
(30/30)	26.3 ±
0.32
(1.2%)	100.0%
(30/30)	26.0 ±
0.12
(0.5%)	100.0%
(30/30)	25.8 ±
0.36
(1.4%)	100.0%
(90/90)	96% -
100%
	Totala	100.0%
(30/30)	N/A	100.0%
(30/30)	N/A	100.0%
(30/30)	N/A	100.0%
(90/90)	96% -
100%
Total	S gene	98.3% (118/120)	95.0% (114/120)	99.2% (119/120)	97.5%
(351/360)	95% -
99%
	ORF1ab	97.5% (117/120)	99.2% (119/120)	100.0% (120/120)	98.9%
(356/360)	97% -
100%
	Totala	100.0% (120/120)	99.2% (119/120)	100.0% (120/120)	99.7%
(359/360)	98% -
100%

"Total based on algorithm that at least one target (S gene or ORF lab) is Detected for the sample to be Detected for SARS-CoV-2.

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

The Limit of Detection (LoD) for NPS and NS was determined to be the lowest detectable concentration of inactivated viral particles (copies/mL) for strain 2019-nCoV/USA-WA1/2020 at which ≥ 95% of all replicates tested positive. Initially, the tentative LoD was identified with serial dilutions of the inactivated viral particles in five (5) replicates for NPS matrix in UTM or four (4) replicates for NS matrix in UTM. The lowest concentration at which all replicates were positive was interpreted as the tentative LoD. For NPS, the LoD was then confirmed by testing forty (40) replicates with concentrations at the tentative LoD while for NS, the LoD was confirmed by testing twenty (20) replicates with concentrations at the tentative LoD. For both NPS and NS, the final LoD was confirmed to be the lowest concentration resulting in positive detection with a minimum 95% positivity. Table 5 shows the results of the studies.

The final LoD for NPS, according to the assay results interpretation was 500 copies/mL. The LoD for NS, according to the assay results interpretation was 242 copies/mL.

9

Image /page/9/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a DNA helix-like graphic on the left, colored in shades of green and blue. To the right of the graphic are the words "DiaSorin" in a dark blue, sans-serif font, with the word "Molecular" underneath in a lighter green color.

Simplexa™ COVID

Table 5: Analytical Sensitivity/Limit of Detection for Inactivated Viral Particles in NPS or NS Matrix in UTM

| Specimen
Type | SARS-CoV-2
LoD
(copies/mL) | % Detection
(# Detected / # Tested) | S gene (FAM)
% Detection
(# Detected/#
Tested) | Mean Ct ± SD
(%CV) | ORF1ab (JOE)
% Detection
(# Detected/#
Tested) | Mean Ct ± SD
(%CV) |
|------------------|----------------------------------|----------------------------------------|---------------------------------------------------------|-----------------------|---------------------------------------------------------|-----------------------|
| NPS | 500 | 100% (40/40)
a | 90% (36/40) | 32.5 ± 1.60
(4.9%) | 100% (40/40) | 31.6 ± 1.56
(4.9%) |
| NS | 242 | 100% (20/20)
a | 80% (16/20) | 34.0 + 0.83
(2.4%) | 80% (16/20) | 32.9 + 0.75
(2.3%) |

Ct = Cycle threshold, SD = Standard Deviation, %CV = Percent Coefficient of Variation

ª % Detection based on algorithm that at least one target (S gene or ORF1ab) is Detected for SARS-CoV-2.

The Limit of Detection (LoD) using inactivated SARS-CoV-2 WHO International Standard viral particles in NPS matrix was determined. The tentative LoD was identified with serial dilutions of the viral particles. The final LoD was confirmed to be the lowest concentration quantified in International Units per milliliter (U/mL) resulting in positive detection with a minimum 95% positivity. Table 6 shows the results of the study.

The final LoD for NPS, according to the assay results interpretation was 500 IU/mL..

		Table 6: Analytical Sensitivity/Limit of Detection for SARS-CoV-2 WHO International Standard

| SARS-CoV-2
WHO
International
Standard
(IU/mL) | % Detection (#
Detected / # Tested) | S gene (FAM) | | ORF1ab (JOE) | |
|-----------------------------------------------------------|----------------------------------------|-------------------------------------------|-----------------------------------------------|-------------------------------------------|--------------------------------------------|
| | | % Detection (#
Detected / #
Tested) | Mean Ct $ \pm $ SD
(%CV) | % Detection (#
Detected / #
Tested) | Mean Ct $ \pm $ SD (%CV) |
| 500 | 97.5% (39/40) | 95% (38/40) | 33.2 $ \pm $ 1.28 (3.8%) | 85% (34/40) | 33.6 $ \pm $ 1.36 (4.1%) |

Ct = Cycle threshold, SD = Standard Deviation, %CV = Percent Coefficient of Variation, IU = International Units

FDA SARS-CoV-2 Reference Panel Testing: The evaluation of sensitivity and MERS-CoV cross-reactivity was performed using reference material (T1), testing blinded samples and according to a standard protocol provided by the FDA. The study included a range finding study and a confirmatory study for the Limit of Detection (LoD). Blinded sample testing was used to establish specificity and to confirm the LoD. The results are summarized in Table 7.

	Table 7: Summary of LoD Confirmation Result using the FDA SARS-CoV-2 Reference Panel

Reference Materials Provided by FDA	Specimen Type	Product LoD	Cross-Reactivity
SARS-CoV-2	NPS	6 x 103 NDU/mL	N/A
MERS-CoV		N/A	ND

NDU/mL = RNA NAAT detectable units/mL, N/A = Not applicable, ND = Not detected, NPS = Nasopharyngeal Swab

10

Image /page/10/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in dark blue, with the word "Molecular" underneath in light green.

Image /page/10/Picture/1 description: The image shows the text 'K212147' in a bold, sans-serif font. The characters are large and fill most of the frame. The color of the text is black, and the background is white, providing a high contrast.

Simplexa™ COVID ge 8 of 13

REACTIVITY/INCLUSIVITY

In Silico

An in silico inclusivity analysis of the COVID-19 target primers and probes in the Simplexa™ COVID-19 Direct assay was performed. All primer and probe sets designed for detection of the ORF1ab and S gene were tested against the complete SARS-CoV-2 genome sequences available in the GISAID database submitted from November 01, 2021 to January 31, 2022. The analysis included 2,170,584 sequences in the amplicon regions of the ORF1ab and S gene primer/probe regions. Only target sequences with full coverage of all three ORF1ab and S gene forward and reverse primer as well as probe region were included in the analyses. The analysis showed that the Simplexa™ COVID-19 Direct target regions had no mismatch to 2,170,382 sequences (~99.99%) and were predicted to be detected by the assay based on sequence homology. There were 202 sequences (~0.01%) with mismatches in at least one primer or probe binding region, region in either ORF1ab or S gene target region. A melting temperature (Tm) analysis was conducted for those sequences with mismatches in the binding sites of both gene assay target regions. A Tm calculation was performed with assay-specific conditions using a Tm Mismatch Bioinformatics Tool. Tm values observed above their respective annealing temperature had mismatches that were not located at the 3' end for the primers; as such, detection of these sequences is not affected by the mismatches.

An additional in silico inclusivity analysis was performed for complete SARS-CoV-2 genome sequences available in the GISAID database submitted from February 01, 2022 to April 30, 2022 including sequences of the Omicron BA.4 and BA.5 subvariants. The analysis included 377,668 sequences in the amplicon regions of the ORF1ab and S gene primer/probe regions. Only target sequences with full coverage of all three ORF1ab and S gene forward and reverse primer as well as probe region were included in the analyses. The analysis showed that the Simplexa™ COVID-19 Direct target regions had no mismatch to 372,411 sequences (~98.6%) and were predicted to be detected by the assay based on sequence homology. There were 5213 (~1.4%) sequences with no mismatches for one gene oligo set (either ORF1ab or S gene), and there were 44 sequences (~0.01%) with mismatches in at least one primer or probe binding region, region in either ORF1ab or S gene target region. A Tm analysis was conducted as described above and the results demonstrated that detection of these sequences is not affected by the mismatches. Table 8 below summarizes the Tm analysis results.

An additional in silico inclusivity analysis was performed for complete SARS-CoV-2 genome sequences available in the GISAID database submitted from May 01, 2022 to July 31, 2022 including sequences of the Omicron BA.2.12.1, BA.2.75, BA.4 and BA.5 subvariants. The analysis included 211,224 sequences in the amplicon regions of the ORF1ab and S gene primer/probe regions. Only target sequences with full coverage of all three ORF1ab and S gene forward and reverse primer as well as probe region were included in the analyses. The analysis showed that the Simplexa™ COVID-19 Direct target regions had no mismatch to 208,582 sequences (~98.7%) and were predicted to be detected by the assay based on sequence homology. There were 2602 (~1.2%) sequences with no mismatches for one gene oligo set (either ORF1ab or S gene), and there were 40 sequences (~0.02%) with mismatches in at least one primer or probe binding region, region in either ORF1ab or S gene target region. A Tm analysis was conducted as described above and the results demonstrated that detection of these sequences is not affected by the mismatches. Table 8 below summarizes the Tm analysis results.

11

Image /page/11/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in a dark blue, sans-serif font, with the word "Molecular" underneath in a lighter green color. The overall design is clean and modern.

510(k) S Simplexa™ COVID-19 Direct September Page 9 of 13

Table 8. Summary of Tm Analysis Results
Timeframe of Sequences
Analyzed	Number of
accessions in GISAID
Database for the
timeframe	Number of sequences
where at least one
target oligo set meets
Tm criteria	Identity to SARS-CoV-
2 gene design
Nov. 1, 2021 to Jan. 31, 2022	2,170,584	2,170,584	100%
Feb. 1, 2022 to Apr. 30, 2022	377,668	377,668	100%
May 1, 2022 to July 31, 2022	211,224	211,224	100%

Wet testing

Analytical reactivity was evaluated using five (5) strains of SARS-CoV-2 viral particles that were available. All five (5) strains of SARS-CoV-2 (Hong Kong/VM200001061/2020, England/204820464/2020, South Africa/KRISP-EC-K005325/2020, Japan/TY7-503/2021, hCoV19/USA/PHC658/2021) were detected at a concentration of 1000 copies/mL All replicates of the test samples had a result of "Detected" according to the interpretation algorithm.

Table 9. Analytical Reactivity Results

| COVID-19 Strain | Tested Concentration | COVID-19 Qualitative Results
% Detection (# Detected
/#Tested) |
|---------------------------------------|----------------------|----------------------------------------------------------------------|
| Hong Kong/VM200001061/2020 | 1000 copies/mL | 100% (3/3) |
| England/204820464/2020 | 1000 copies/mL | 100% (3/3) |
| South Africa/KRISP-ECK005325/
2020 | 1000 copies/mL | 100% (3/3) |
| Japan/TY7-503/2021 | 1000 copies/mL | 100% (3/3) |
| hCoV19/USA/PHC658/2021 | 1000 copies/mL | 100% (3/3) |

CROSS-REACTIVITY

Cross-reactivity of the Simplexa™ COVID-19 Direct assay was evaluated using both in silico analysis and by testing whole organisms or purified nucleic acid from other organisms. Analytical specificity/crossreactivity was tested using 47 different viruses, bacteria, and fungi that could be found in nasopharyngeal swab specimens. In addition, pooled human nasal fluid was tested to represent diverse microbial flora. Test specimens for laboratory testing were prepared by spiking cultured isolates/inactivated organisms/purified nucleic acids (whole genome) (i.e., a minimum of 10° CFU/mL or higher for bacteria and typically 105 TCIDsolmL or PFU/mL or higher for viruses) into COVID-19 negative pooled NPS matrix collected in UTM All samples were tested in three replicates. The results from the cross-reactivity testing are summarized in Table 10.

| Organism Concentration1 (# Detected/#Tested) | |------------------------ | (FAM) (JOE) | IC
(Q670) |
| Adenovirus C (Type 1) | Adenovirus 7A | Bordetella pertussis | Candida albicans | Chlamydophila pneumoniae | Chlamydophila | | Organism | (FAM) (JOE) | IC
(Q670) |
| Corynebacterium diphtheriae | Coxiella burnetii (genomic DNA) | Cytomegalovirus | Enterovirus 68 | Epstein-Barr virus | Escherichia coli | Haemophilus influenzae | Human coronavirus 229E* | Human coronavirus HKU1 (RNA) copies/mL | 0.0% (0/3) | Human coronavirus NL63* | Human coronavirus OC43 | Human genomic DNA (Leukocytes) | Human metapneumovirus (hMPV)* | Influenza A/Perth/16/2009 | Influenza B/Florida/02/06 | Lactobacillus plantarum 17-5 | Legionella longbeachae | Legionella pneumophila | Leptospira interrogans | Measles | MERS-coronavirus | Moraxella catarrhalis | Mumps | Mycobacterium DNA) | 1 x 106 copies/mL | Mycoplasma pneumoniae | Neisseria elongata | Neisseria meningitidis | Parainfluenza virus 1 | Parainfluenza virus 2 | Parainfluenza virus 3 | Parainfluenza virus 4 | Parechovirus 3 | Pseudomonas aeruginosa | Respiratory syncytial Virus A | Respiratory syncytial Virus B | Rhinovirus | SARS-coronavirus (RNA) | Staphylococcus aureus | Staphylococcus epidermidis | Streptococcus pneumoniae | Streptococcus pyogenes | Streptococcus salivarius | Pooled Human Nasal Fluid | Tested
| Qualitative Results: % Detection
| |
---------------------|-----------------------------|----------------------------------------------------------|-----------------|--------------|
| | S gene
| ORF1ab
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 TCID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 IFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
psittaci (genomic DNA) | 1 x 106 copies/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| Tested | Qualitative Results: % Detection | | |
| | (# Detected/#Tested) | | |
| Concentration1 | S gene
| ORF1ab
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 copies/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 copies/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 3 x 104 TCID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 genome
| 0.0% (0/3) | 100.0% (3/3) |
| 3 x 104 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 TCID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 cells/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 3 x 104 TCID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 EID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1:10 Dilution | 0.0% (0/6) | 0.0% (0/6) | 100.0% (6/6) |
| 1 x 105 TCID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 TCID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
tuberculosis (genomic
| 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CCU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 TCID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 TCID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 TCID50/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 U/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 105 copies/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1 x 106 CFU/mL | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |
| 1:1 Dilution | 0.0% (0/3) | 0.0% (0/3) | 100.0% (3/3) |

Table 10: Laboratory Tested Cross-Reactivity Analysis

12

Image /page/12/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in dark blue on the top line and "Molecular" in green on the bottom line.

510(k) Summary Simplexa™ COVID-19 Direct MOL4150 September 12, 2022

Page 10 of 13

*A lower concentration was tested due to inability to obtain stock material with high titer

1CCU/mL = Color Changing Units/milliter, CFU/mL = Colony Forming Units/milliliter, IFU/mL = Infectious units/milliliter, U/mL = Units/milliliter, TCID50/mL = Tissue Culture Infectious Dose/milliliter

Bacillus anthracis, Influenza C and Pneumocystis jirovecii were not available. In addition, human coronavirus 229E, human coronavirus NL63 and human metapneumovirus (hMPV) were not available at a high enough concentration to permit wet testing at the concentration levels stated above. As a result of in silico BLAST analysis, the assay was found not to cross-react with these organisms.

13

Image /page/13/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a stylized DNA double helix on the left, rendered in shades of green and blue. To the right of the helix is the company name, "DiaSorin," in a bold, dark blue font. Below "DiaSorin" is the word "Molecular" in a lighter green color, completing the logo.

POTENTIAL INTERFERING SUBSTANCES

Potential interfering substances from respiratory specimens were tested for ability to generate false negative results. The test samples contained inactivated SARS-CoV-2 at a concentration of 3x LoD in pooled negative nasopharyngeal swab matrix. Testing was performed with three (3) replicates per substance. The FluMist nasal vaccine was not tested as an interfering substance since it was unavailable at the time of the study.

| Potentially Interfering
Substance | Active
Ingredient | Tested
Concentration | COVID-19
Qualitative
Results: %
Detection
(# Detected
/#Tested) | IC Qualitative
Results: %
Detection
(# Detected
/#Tested) |
|---------------------------------------------------------------|-------------------------------------------------------------------------------------------|-------------------------|--------------------------------------------------------------------------------|-----------------------------------------------------------------------|
| Antibiotic nasal ointment
(Mupirocin) | Mupirocin | 6.6 mg/mL | 100.0% (3/3) | 100.0% (3/3) |
| Anti-viral drug (Oseltamivir) | Oseltamivir | 3.3 mg/mL | 100.0% (3/3) | 100.0% (3/3) |
| Cold Eeze (Throat lozenges,
Oral anesthetic and analgesic) | Zincum
gluconicum 2X | 2.5% (w/v) | 100.0% (3/3) | 100.0% (3/3) |
| Homeopathic allergy relief
medicine | N/A | 10% (v/v) | 100.0% (3/3) | 100.0% (3/3) |
| Mucin (Bovine submaxillary gland,
type I-S) | N/A | 5 mg/mL | 100.0% (3/3) | 100.0% (3/3) |
| Nasal corticosteroids (Fluticasone) | Fluticasone | 5% (v/v) | 100.0% (3/3) | 100.0% (3/3) |
| Allergy Relief Swabs (Nasal Gel,
Zicam) | Luffa
opperculata,
Galphimia
glauca,
histaminum
hydrochloricum,
Sulphur | 5% (w/v) | 100.0% (3/3) | 100.0% (3/3) |
| Nasal spray or drops
(Oxymetazoline) | Oxymetazoline | 15% (v/v) | 100.0% (3/3) | 100.0% (3/3) |
| Saliva | N/A | 10% (v/v) | 83.3% (5/6) | 83.3% (5/6) |
| | N/A | 5% (v/v) | 100.0% (6/6) | 100.0% (6/6) |
| Systemic antibacterial
(Tobramycin) | Tobramycin | 4 µg/mL | 100.0% (3/3) | 100.0% (3/3) |
| Whole Blood | N/A | 2% (v/v) | 100.0% (3/3) | 100.0% (3/3) |
| Zanamivir | N/A | 3 mg/mL | 100.0% (6/6) | 83.3% (5/6) |

Table 11: Potential Interfering Substances

*Interference from saliva was observed at a concentration above 5%

mg = milligram, mL = milliliter, v/y = volume, w/v = weight to volume, ug = microgram, NA = Not Applicable

INTERFERENCE BY OTHER MICROORGANISMS

The Simplexa™ COVID-19 Direct assay was evaluated by testing the ability to identify SARS-CoV-2 when other potentially inhibitory organisms were present. The panel of forty-seven (47) potentially inhibitory organisms was individually spiked into a pool with a low concentration at two times (2x) LoD of inactivated COVID-19 viral particles in NPS matrix. In addition, pooled human nasal fluid was tested to represent diverse microbial flora. For organisms not titered in CFU/mL, other industry acceptable units were used as indicated. Samples were assayed in triplicate to screen for potential inhibition. No inhibition by other organisms was observed at the concentrations indicated in Table 12. Lactobacillus plantarum 17-5 showed interference when tested above (5 x 105 CFU/mL).

14

Image /page/14/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in dark blue on the top line and "Molecular" in green on the bottom line.

510(k) Summary Simplexa™ COVID-19 Direct MOL4150 September 12, 2022 Page 12 of 13

| Organism | Tested
Concentration¹ | COVID-19 Qualitative
Results: % Detection
(# Detected /#Tested) |
|------------------------------------------|--------------------------|-----------------------------------------------------------------------|
| Adenovirus C (Type 1) | 1 x 10⁵ U/mL | 100.0% (3/3) |
| Adenovirus 7A | 1 x 10⁵ TCID₅₀/mL | 100.0% (3/3) |
| Bordetella pertussis | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Candida albicans | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Chlamydia pneumoniae | 1 x 10⁶ IFU/mL | 100.0% (3/3) |
| Chlamydophila psittaci (genomic DNA) | 2.5 x 10⁵ copies/mL* | 100.0% (3/3) |
| Corynebacterium diphtheriae | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Coxiella burnetii (genomic DNA) | 1 x 10⁶ copies/mL | 100.0% (3/3) |
| Cytomegalovirus | 1 x 10⁵ U/mL | 100.0% (3/3) |
| Enterovirus 68 | 1 x 10⁵ U/mL | 100.0% (3/3) |
| Epstein-Barr virus | 1 x 10⁵ copies/mL | 100.0% (3/3) |
| Escherichia coli | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Haemophilus influenzae | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Human coronavirus 229E | 1.5 x 10⁴ TCID₅₀/mL* | 100.0% (3/3) |
| Human coronavirus HKU1 (RNA) | 1 x 10⁵ genome copies/mL | 100.0% (3/3) |
| Human coronavirus NL63 | 1.5 x 10⁴ U/mL* | 100.0% (3/3) |
| Human coronavirus OC43 | 1 x 10⁵ TCID₅₀/mL | 100.0% (3/3) |
| Human genomic DNA (Leukocytes) | 1 x 10⁶ cells/mL | 100.0% (3/3) |
| Human metapneumovirus (hMPV) | 1.5 x 10⁴ TCID₅₀/mL* | 100.0% (3/3) |
| Influenza A/Perth/16/2009 | 1 x 10⁵ EID₅₀/mL | 100.0% (3/3) |
| Influenza B/Phuket/3073/2013 | 1 x 10⁵ CEID₅₀/mL | 100.0% (3/3) |
| Lactobacillus plantarum 17-5** | 5 x 10⁵ CFU/mL | 100.0% (6/6) |
| Legionella longbeachae | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Legionella pneumophila | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Leptospira interrogans | 1:10 Dilution | 100.0% (6/6) |
| Measles | 1 x 10⁵ TCID₅₀/mL | 100.0% (3/3) |
| MERS-coronavirus | 1 x 10⁵ TCID₅₀/mL | 100.0% (3/3) |
| Moraxella catarrhalis | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Mumps | 1 x 10⁵ U/mL | 95.0% (19/20) |
| Mycobacterium tuberculosis (genomic DNA) | 1 x 10⁶ copies/mL | 100.0% (3/3) |
| Mycoplasma pneumoniae | 1 x 10⁶ CCU/mL | 100.0% (3/3) |
| Neisseria elongata | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Neisseria meningitidis | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Parainfluenza virus 1 | 1 x 10⁵ U/mL | 100.0% (3/3) |
| Parainfluenza virus 2 | 1 x 10⁵ U/mL | 100.0% (3/3) |
| Parainfluenza virus 3 | 1 x 10⁵ TCID₅₀/mL | 100.0% (3/3) |
| Parainfluenza virus 4 | 1 x 10⁵ U/mL | 100.0% (3/3) |
| Parechovirus 3 | 1 x 10⁵ U/mL | 100.0% (3/3) |
| Pseudomonas aeruginosa | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Respiratory syncytial virus A | 5 x 10⁴ TCID₅₀/mL* | 100.0% (3/3) |
| Respiratory syncytial virus B | 1 x 10⁵ TCID₅₀/mL | 100.0% (3/3) |
| Rhinovirus | 1 x 10⁵ U/mL | 100.0% (3/3) |
| SARS-Coronavirus (RNA) | 1 x 10⁵ copies/mL | 100.0% (3/3) |
| Staphylococcus aureus | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Staphylococcus epidermidis | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Streptococcus pneumoniae | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Streptococcus pyogenes | 1 x 10⁶ CFU/mL | 100.0% (3/3) |
| Streptococcus salivarius | 1 x 10⁶ CFU/mL | 100.0% (3/3) |

Table 12. Simplexa™ COVID-19 Direct – Microbial Interference

*A lower concentration was tested due to inability to obtain stock material with high titer

**Interfernce with Lactobacillus plantarum 17-5 was observed at a concentration above 5 x 105 CFU/mL.

1CCU = Color changing units/milliliter, CFU/mL = Colony forming units/milliiter, IFU/ml = Infectious units/milliliter, U/mL = Units/milliliter, TCID50/mL = Tissue Culture Infectious Dose per milliliter

15

Image /page/15/Picture/0 description: The image contains the logo for DiaSorin Molecular. The logo consists of a stylized DNA helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in dark blue on the top line and "Molecular" in green on the bottom line.

CARRY-OVER CONTAMINATION

Amplification carry-over for the Simplexa™ assays has been assessed against existing assays that use the same sample matrices, workflow and specimen type, and therefore no carry-over is anticipated. The study was designed by alternately placing high positive and negative samples on each disc. No evidence of carry-over contamination was observed.

Conclusion

The analytical and method comparison studies have demonstrated that the Simplexa™ COVID-19 Direct is Substantially Equivalent to the predicate device (K211079). The device labeling is compliant with 21 CFR ട്ട 809.10.