The DiaSorin Molecular Simplexa™ COVID-19 & Flu A/B Direct is a real-time RT-PCR assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection and differentiation of nucleic acid from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza B virus in nasopharyngeal swabs (NPS) from individuals with signs and symptoms of respiratory tract infection.

The Simplexa™ COVID-19 & Flu A/B Direct assay is intended for use as an aid in the differential diagnosis of SARS-CoV-2, influenza A and influenza B infection.

Negative results do not preclude SARS-CoV-2, influenza B infection and should not be used as the sole basis for patient management decisions. Positive results do not rule out coinfection with other organisms. Results should be combined with clinical observations, patient history, and epidemiological information.

The Simplexa™ COVID-19 & Flu A/B Direct assay is intended for use by qualified and trained clinical laboratory personnel specifically instructed and trained in the techniques of real-time PCR and in vitro diagnostic procedures.

Device Description

The Simplexa™ COVID-19 & Flu A/B Direct assay system is a real-time RT-PCR system that enables the direct amplification, detection and differentiation of SARS-CoV-2 RNA, human influenza A (Flu A) virus RNA and human influenza B (Flu B) virus RNA from unprocessed nasopharyngeal swabs (NPS) that have not undergone nucleic acid extraction. The system consists of the Simplexa™ COVID-19 & Flu A/B Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc and associated accessories.

In the Simplexa™ COVID-19 & Flu A/B Direct assay, fluorescent probes are used together with corresponding forward and reverse primers to amplify SARS-CoV-2, Flu A, Flu B and internal control RNA targets. For COVID-19 detection, the assay targets two different regions specific to the SARS-CoV-2 genome; the S gene which encodes the spike glycoprotein and the ORF1ab region which encodes wellconserved non-structural proteins and therefore is less susceptible to recombination. For Flu detection the assay targets conserved regions of influenza A viruses (matrix gene) and influenza B viruses (matrix gene). The assay provides three results; COVID-19 (ORF1ab and/or S gene detection), influenza A viruses (matrix gene detection) and influenza B viruses (matrix gene detection). An RNA internal control is used to detect RT-PCR failure and/or inhibition.

AI/ML Overview

This document describes the analytical and clinical performance studies for the DiaSorin Molecular Simplexa™ COVID-19 & Flu A/B Direct assay.

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria for the clinical performance are generally indicated by the Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) with their corresponding 95% Confidence Intervals (CI).

Target	Acceptance Criteria (95% CI) (Implicit, based on study results)	Reported Device Performance (Prospective Study, PPA/NPA)	Reported Device Performance (Retrospective Study, PPA/NPA)
Influenza A	PPA: >82.5%, NPA: >99.3%	PPA: 91.9% (57/62), 95% CI: 82.5% - 96.5%NPA: 99.8% (1104/1106), 95% CI: 99.3% - 100%	PPA: 97.6% (80/82), 95% CI: 91.5% - 99.3%NPA: 100% (176/176), 95% CI: 97.9% - 100%
Influenza B	PPA: N/A (for prospective), NPA: >99.7%	PPA: N/A (0/0 occurrences)NPA: 100% (1165/1165), 95% CI: 99.7% - 100%	PPA: 98.2% (112/114), 95% CI: 93.8% - 99.5%NPA: 100% (144/144), 95% CI: 97.4% - 100%
SARS-CoV-2	PPA: >92.1%, NPA: >95.5%	PPA: 98.5% (67/68), 95% CI: 92.1% - 99.7%NPA: 97.4% (417/428), 95% CI: 95.5% - 98.6%	PPA: N/A (0/0 occurrences)NPA: 100% (252/252), 95% CI: 98.5% - 100%

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

Prospective Samples: Over 1400 total specimens (nasopharyngeal swabs (NPS)) were collected between August 2021 and March 2022 from six geographically diverse clinical sites within the United States. The exact number of prospective samples used for each target in the agreement analysis can be inferred from the TP/(TP+FN) and TN/(TN+FP) values in Table 2:
- Influenza A: 1168 (62 positive, 1106 negative, plus 5 and 2 discrepant)
- Influenza B: 1165 (all negative)
- SARS-CoV-2: 496 (68 positive, 428 negative, plus 1 and 11 discrepant)
Retrospective Samples: 82 positive influenza B specimens and 62 negative specimens were used. These were blinded and randomized for the study. The exact number of retrospective samples used for each target in the agreement analysis can be inferred from the TP/(TP+FN) and TN/(TN+FP) values in Table 3:
- Influenza A: 258 (82 positive, 176 negative)
- Influenza B: 258 (114 positive, 144 negative)
- SARS-CoV-2: 252 (all negative)

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 or their specific qualifications (e.g., radiologist with 10 years of experience) used to establish the ground truth for the test set.

4. Adjudication Method for the Test Set

SARS-CoV-2: For SARS-CoV-2, a composite reference method (CRM) was used. This involved three COVID-19 Emergency Use Authorized (EUA) NAAT assays. The adjudication method was:
- "Detected" CRM if two out of three EUA assays were positive.
- "Not Detected" CRM if two out of three EUA assays were negative.
Influenza A and B: For influenza A and B, the comparator was an FDA-cleared NAAT. There is no mention of a multi-assay composite reference method, suggesting a single FDA-cleared NAAT was used as the ground truth. Discrepancy analysis involved additional FDA cleared NAATs and PCR followed by BDS.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was Done

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done. This study focuses on the standalone performance of the diagnostic assay rather than human reader performance with or without AI assistance.

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) was Done

Yes, a standalone performance study was done. The entire clinical performance section evaluates the Simplexa™ COVID-19 & Flu A/B Direct assay's ability to detect and differentiate nucleic acids directly, without human interpretation in the results reporting, other than laboratory personnel operating the instrument. The results are presented as the assay's agreement with reference methods.

7. The Type of Ground Truth Used

SARS-CoV-2: Composite reference method (CRM) based on the consensus of three COVID-19 Emergency Use Authorized (EUA) NAAT assays.
Influenza A and B: An FDA-cleared NAAT was used as the primary comparator. In cases of discrepancy, additional FDA-cleared NAATs and PCR followed by Bidirectional Sequencing (BDS) were used for confirmation.

8. The Sample Size for the Training Set

The document describes the clinical performance (test set) and analytical studies. It does not explicitly mention a "training set" in the context of machine learning. The assay is a real-time RT-PCR assay, which typically relies on pre-defined primer and probe sequences rather than a machine learning model that requires a distinct training set in the conventional sense. The development and optimization of the primer/probe sets (e.g., analytical reactivity, inclusivity) can be considered analogous to a "training" or development phase, but no specific dataset labeled as such is provided.

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

As noted in point 8, a traditional "training set" with ground truth establishment in the context of machine learning is not applicable to this RT-PCR assay. The analytical studies (Limit of Detection, Analytical Reactivity/Inclusivity, Cross-Reactivity, Interfering Substances, Competitive Interference, Microbial Interference) use quantitated viral stocks, cultured isolates, purified nucleic acids, or in silico analysis against public strain databases (e.g., GISAID) to demonstrate the assay's analytical performance across a wide range of relevant targets and conditions. This ensures the assay's biochemical design (primers, probes) is sound and effective.

Summary

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March 17, 2023

Image /page/0/Picture/1 description: The image contains 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 Sr. Director, Molecular Regulatory Affairs 11331 Valley View Street Cypress, California 90630

Re: K220963

Trade/Device Name: Simplexa COVID-19 & Flu A/B 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: OOF, OOI Dated: March 30, 2022 Received: April 1, 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.

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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 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 -S

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

Enclosure

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Indications for Use

510(k) Number (if known) K220963

Device Name Simplexa™ COVID-19 & Flu A/B Direct

Indications for Use (Describe)

The Simplexa™ COVID-19 & Flu A/B Direct assay is intended for use as an aid in the differential diagnosis of SARS-CoV-2, influenza A and influenza B infection.

Type of Use ( Select one or both, as applicable )
☒ 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 Simplexa™ COVID-19 & Flu A/B L Page 1 of 18

Applicant	DiaSorin Molecular LLC.11331 Valley View StreetCypress, California 90630USA
Establishment Registration No.	2023365
Contact Person	Tara VivianiSr. Director Molecular Regulatory AffairsTel. 562.240.6500Tara.Viviani@DiaSorin.com
Summary Date	March 16, 2023
Proprietary Name	Simplexa™ COVID-19 & Flu A/B Direct
US Product Codes/Names andRegulation Numbers	QOF - Multi-Target Respiratory Specimen Nucleic Acid TestIncluding Sars-Cov-2 And Other Microbial Agents 21 CFR §866.3981OOI - Real Time Nucleic Acid Amplification System 21 CFR §862.2570
Classification	Class II
Predicate Devices	BioFire Respiratory Panel 2.1 (RP2.1) (DEN200031)

Intended Use

Simplexa COVID-19 & Flu A/B Direct (Catalog Number: MOL4250):

The Simplexa™ COVID-19 & Flu A/B Direct assay is intended for use as an aid in the differential diagnosis of SARS-CoV-2, influenza A and influenza B infection.

Device Description

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510(k) Si marv Simplexa™ COVID-19 & Flu A/B Direct I OL4250 March 16. 2023 Page 2 of 18

not undergone nucleic acid extraction. The system consists of the Simplexa™ COVID-19 & Flu A/B Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc and associated accessories.

Simplexa™ COVID-19 &Flu A/B Direct REF MOL4250

Component Name	REF	EC Symbol on Label	Abbreviated Name	Cap Color	Number of Vials	Reactions per Vial/Kit	Volume per Vial
Simplexa ™ COVID-19& Flu A/B Direct Reaction Mix	MOL4251	REAG C	RM	Blue	24	1/24	50 μL

Simplexa™ COVID-19 &Flu A/B Direct Components and Descriptions

KitComponent	Contents
	DNA polymerase, Reverse Transcriptase, RNase inhibitor, buffer and dNTPs, encapsulated RNATemplate, primers and dye-labeled fluorescent probes specific for detection of SARS-CoV-2,influenza A, influenza B and for the RNA Internal Control
	Target	Channel	Excitation	Emission	Targeted Gene
Simplexa™COVID-19 &Flu A/B DirectReaction Mix(RM)	Flu A	520	445-505	507-533	matrix
	Flu B	560	505-543	547-573	matrix
	SARS-CoV-2"COVID"	610	502-596	597-623	S and ORF1ab
	Internal Control"RNA IC"	690	622-658	652-708	N/A
Simplexa™COVID-19 &Flu A/B DirectBarcode Card	Assay specific parameters, lot number, expiration date

Materials Supplied Separately

Direct Amplification Disc Kit (REF MOL1455) Direct Amplification Discs for use on the LIAISON® MDX Simplexa™ COVID-19 & Flu A/B Positive Control Pack (REF) MOL2260)

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510(k) Summary

March 16, 2023 Page 3 of 18

Comparison to Predicate Device

Comparison toPredicate Device	Predicate DeviceBioFire Respiratory Panel 2.1 (RP2.1)(DEN200031)	Candidate Device:
Product Code	QOF	QOF
RegulationNumber	21 CFR 866.3981	21 CFR 866.3981
OrganismDetected	Severe Acute Respiratory SyndromeCoronavirus (SARS-CoV-2), Adenovirus,Coronavirus 229E, Coronavirus HKU1,Coronavirus NL63, Coronavirus OC43,Human Metapneumovirus, HumanRhinovirus/Enterovirus, Influenza A,including subtypes H1, H1-2009, and H3,Influenza B, Parainfluenza Virus 1,Parainfluenza Virus 2, Parainfluenza Virus 3,Parainfluenza Virus 4, Respiratory SyncytialVirus, Bordetella parapertussis (IS1001),Bordetella pertussis (ptxP), Chlamydiapneumoniae, and Mycoplasma pneumoniae	Severe Acute Respiratory SyndromeCoronavirus (SARS-CoV-2), Influenza A (FluA) and Influenza B (Flu B)
Measurand	DNA/RNA	RNA
Target	FilmArray Torch TechnologySARS-CoV-1 Well conserved region of the SgeneSARS-CoV-2 Well conserved region of theM gene	SARS-CoV-2 Well conserved regions of theS and ORF genesFlu A and B Targets Well conserved regionsof the matrix gene (TAQ Man Technology)
Intended Use Kit	The BioFire Respiratory Panel 2.1 (RP2.1) isa PCR-based multiplexed nucleic acid testintended for use with the BioFire FilmArray2.0 or BioFire FilmArray Torch systems forthe simultaneous qualitative detection andidentification of multiple respiratory viral andbacterial nucleic acids in nasopharyngealswabs (NPS) obtained from individualssuspected of respiratory tract infections,including COVID-19.The following organism types and subtypesare identified using the BioFire RP2.1:● Adenovirus,● Coronavirus 229E,● Coronavirus HKU1,● Coronavirus NL63,● Coronavirus OC43,● Severe Acute RespiratorySyndrome● Coronavirus (SARS-CoV-2),● Human Metapneumovirus,● Human Rhinovirus/Enterovirus,● Influenza A, including subtypes H1,H1-2009, and H3,● Influenza B,● Parainfluenza Virus 1,● Parainfluenza Virus 2,● Parainfluenza Virus 3,● Parainfluenza Virus 4,	The DiaSorin Molecular Simplexa™ COVID-19 & Flu A/B Direct is a real-time RT-PCRassay intended for use on the LIAISON® MDXinstrument for the in vitro qualitative detectionand differentiation of nucleic acid from severeacute respiratory syndrome coronavirus 2(SARS-CoV-2), influenza A virus andinfluenza B virus in nasopharyngeal swabs(NPS) from individuals with signs andsymptoms of respiratory tract infection.The Simplexa™ COVID-19 & Flu A/B Directassay is intended for use as an aid in thedifferential diagnosis of SARS-CoV-2,influenza A and influenza B infection.Negative results do not preclude SARS-CoV-2, influenza A or influenza B infection andshould not be used as the sole basis forpatient management decisions. Positiveresults do not rule out coinfection with otherorganisms; Results should be combined withclinical observations, patient history, andepidemiological information.The Simplexa™ COVID-19 & Flu A/B Directassay is intended for use by qualified andtrained clinical laboratory personnelspecifically instructed and trained in thetechniques of real-time PCR and in vitrodiagnostic procedures.
Comparison toPredicate Device	Predicate DeviceBioFire Respiratory Panel 2.1 (RP2.1)(DEN200031)	Candidate Device:
	Bordetella parapertussis (IS1001), Bordetella pertussis (ptxP), Chlamydia pneumoniae, and Mycoplasma pneumoniae Nucleic acids from the respiratory viral andbacterial organisms identified by this test aregenerally detectable in NPS specimensduring the acute phase of infection. Thedetection and identification of specific viraland bacterial nucleic acids from individualsexhibiting signs and/or symptoms ofrespiratory infection is indicative of thepresence of the identified microorganismand aids in the diagnosis of respiratoryinfection if used in conjunction with otherclinical and epidemiological information. Theresults of this test should not be used as thesole basis for diagnosis, treatment, or otherpatient management decisions.Negative results in the setting of arespiratory illness may be due to infectionwith pathogens that are not detected by thistest, or lower respiratory tract infection thatmay not be detected by an NPS specimen.Positive results do not rule out coinfectionwith other organisms. The agent(s) detectedby the BioFire RP2.1 may not be the definitecause of disease. Additional laboratorytesting (e.g. bacterial and viral culture,immunofluorescence, and radiography) maybe necessary when evaluating a patient withpossible respiratory tract infection.	-
AutomatedSystem (Sampleto Answer)	Automated	Same
Instrumentation	BioFire® FilmArray® 2.0 or BioFire®FilmArray® Torch Systems	LIAISON® MDX
Sample Types	Nasopharyngeal Swab (NPS)	Same

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510(k) Summary Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 4 of 18

CLINICAL PERFORMANCE

The performance of Simplexa™ COVID-19 & Flu A/B Direct was evaluated using prospective, archived positive and negative nasopharyngeal swab (NPS) specimens from human patients with signs and symptoms of respiratory tract infection. The clinical study was conducted using a total of over 1400 prospective and archived specimens in transport media. Prospective specimens from pediatric (0 - 99 years) were collected from six (6) geographically diverse clinical sites within the United States between August 2021 and March 2022. The Simplexa™ COVID-19 & Flu A/B Direct clinical agreement testing was performed at two (2) external clinical sites and one (1) internal site. Additionally, retrospective archived specimens consisted of 82 positive influenza B, and 62 negative specimens blinded and randomized for the study. The comparator for influenza A and B targets was an FDA cleared NAAT. For the SARS-CoV-2, target three COVID-19 Emergency Use Authorized NAAT assays were used to establish a composite reference method (CRM). Two out of three positive results determined "Detected" CRM and two out of three negative results determined "Not Detected" CRM.

Table 2 shows the results of the Simplexa™ COVID-19 & Flu A/B Direct assay and comparator assay results in the prospective study analysis and Table 3 shows the retrospective study analysis. The positive percent agreement (PPA) and negative percent agreement (NPA) were calculated for each tarqet.

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510(k) Summarv Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 5 of 18

Table 2. Simplexa™ COVID-19 & Flu A/B Direct Prospective Study Analysis – Aug. 2021 to Mar. 2022

Collection
Simplexa™ COVID-19 & Flu A/B Direct Target	Positive Percent Agreement			Negative Percent Agreement
	TP / (TP+FN)	PPA (%)	95% CI	TN / (TN+FP)	NPA (%)	95% CI
Influenza A	57/62a	91.9%	82.5% - 96.5%	1104/1106b	99.8%	99.3% - 100%
Influenza B	0/0	N/A	N/A	1165/1165	100%	99.7% - 100%
SARS-CoV-2	67/68c	98.5%	92.1% - 99.7%	417/428d	97.4%	95.5% - 98.6%

a Five (5) specimens were negative by an additional FDA cleared NAAT.

' Two (2) specimens were positive by an additional FDA cleared NAAT. One of the two specimens (1/2) was tested with PCR followed by BDS and was positive.

· One specimen was positive by an additional FDA cleared NAAT.

4 Nine of the eleven specimens (9/11) were positive by an additional FDA cleared NAAT and four (4) were positive by PCR followed by BDS. N/A = Not Applicable, PPA = Positive Percent, NPA = Negative Percent, C1 = Confidence Interval. The 95% confidence intervals (CI) were calculated following the Wilson Score method.

				Table 3. Simplexa™ COVID-19 & Flu A/B Direct Retrospective (Archived) Study Analysis
--	--	--	--	--------------------------------------------------------------------------------------	--	--	--

Simplexa™ COVID-19 & FluA/B Direct Target	Positive Percent Agreement			Negative Percent Agreement
	TP /(TP+FN)	PPA (%)	95% CI	TN /(TN+FP)	NPA (%)	95% CI
Influenza A	80/82	97.6%	91.5% - 99.3%	176/176	100%	97.9% - 100%
Influenza B	112/114	98.2%	93.8% - 99.5%	144/144	100%	97.4% - 100%
SARS-CoV-2	0/0	N/A	N/A	252/252	100%	98.5% -100%

N/A = Not Applicable, PPA = Positive Percent, NPA = Neqative Percent Agreement, Cl = Confidence Interval. The 95% confidence intervals (CI) were calculated following the Wilson Score method.

REPRODUCIBILITY

The reproducibility study was performed by one (1) internal sites. The panel consisted of eight (8) reproducibility panel members, including six (6) contrived samples,one (1) negative sample (UTM as No Template Control (NTC)] and one (1) positive control (PC). The contrived panel members were prepared by spiking each analyte at approximately two times the Limit of Detection (2x LoD, low positive) and approximately four times (4x) LoD (medium positive) into native negative nasopharyngeal swab matrix in UTM. Each panel member was tested in triplicate for five (5) days. 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. Agreement with expected results are presented in Table 4 with average Cts, standard deviation (SD) and coefficient of variation (%CV).

					Table 4. Simplexa™ COVID-19 & Flu A/B Direct Reproducibility
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	Site 1		Site 2		Site 3		All Sites
Sample	Agreementwithexpectedresults	Avg. Ct ±SD(%CV)	Agreementwithexpectedresults	Avg. Ct ±SD(%CV)	Agreementwithexpectedresults	Avg. Ct ±SD(%CV)	Agreementwithexpectedresults	Avg. Ct+ SD(%CV)	95% Cl
Flu A Hong Kong -LP	100.0%(30/30)	32.3 ±0.54(1.7%)	100.0%(30/30)	32.9 ±1.22(3.7%)	100.0%(30/30)	32.8 ±0.55(1.7%)	100.0%(90/90)	32.7 ±0.86(2.6%)	95.9% to100.0%
Flu A Hong Kong -MP	100.0%(30/30)	31.7 ±0.49(1.5%)	100.0%(30/30)	31.5 ±0.43(1.4%)	100.0%(30/30)	31.9 ±0.44(1.4%)	100.0%(90/90)	31.7 ±0.47(1.5%)	95.9% to100.0%
Flu B Malaysia -LP	100.0%(30/30)	30.8 ±0.69(2.2%)	100.0%(30/30)	30.9 ±0.60(1.9%)	100.0%(30/30)	31.5 ±0.62(2.0%)	100.0%(90/90)	31.1 ±0.70(2.2%)	95.9% to100.0%
Flu B Malaysia -MP	100.0%(30/30)	30.0 ±0.35(1.2%)	100.0%(30/30)	29.9 ±0.44(1.5%)	100.0%(30/30)	30.4 ±0.48(1.6%)	100.0%(90/90)	30.1 ±0.48(1.6%)	95.9% to100.0%

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510(k) Summary

Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 6 of 18

	Site 1		Site 2		Site 3		All Sites
Sample	Agreementwithexpectedresults	Avg. Ct ± SD(%CV)	Agreementwithexpectedresults	Avg. Ct ± SD(%CV)	Agreementwithexpectedresults	Avg. Ct ± SD(%CV)	Agreementwithexpectedresults	Avg. Ct ± SD(%CV)	95% CI
COVID-19 USA-WA1 - LP	100.0%(30/30)	30.2 ± 0.41(1.4%)	100.0%(30/30)	29.5 ± 0.40(1.4%)	100.0%(30/30)	30.2 ± 0.67(2.2%)	100.0%(90/90)	30.0 ± 0.59(2.0%)	95.9% to100.0%
COVID-19 USA-WA1 - MP	100.0%(30/30)	29.0 ± 0.46(1.6%)	100.0%(30/30)	28.8 ± 0.45(1.6%)	100.0%(30/30)	29.4 ± 0.61(2.1%)	100.0%(90/90)	29.1 ± 0.56(1.9%)	95.9% to100.0%
Positive Control(PC)	100.0%(30/30)	26.2 ± 0.32(1.2%)	100.0%(30/30)	26.1 ± 0.21(0.8%)	100.0%(30/30)	26.0 ± 0.20(0.8%)	100.0%(90/90)	26.1 ± 0.26(1.0%)	95.9% to100.0%
Positive Control(PC)		27.6 ± 0.87(3.2%)		28.2 ± 0.39(1.4%)		27.1 ± 0.22(0.8%)		27.6 ± 0.71(2.6%)
Positive Control(PC)		27.8 ± 1.18(4.2%)		27.2 ± 0.23(0.9%)		26.8 ± 0.22(0.8%)		27.3 ± 0.80(2.9%)
Negative (UTM)	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)	0.0 ± 0.00(N/A%)	95.9% to100.0%
Negative (UTM)		0.0 ± 0.00(N/A%)		0.0 ± 0.00(N/A%)		0.0 ± 0.00(N/A%)		0.0 ± 0.00(N/A%)
Negative (UTM)		0.0 ± 0.00(N/A%)		0.0 ± 0.00(N/A%)		0.0 ± 0.00(N/A%)		0.0 + 0.00(N/A%)
Total	100.0%(240/240)		100.0%(240/240)		100.0%(240/240)		100.0%(720/720)		99.5% to100.0%

Ct. = Cycle threshold

SD = Standard Deviation

%CV = Percent Coefficient of Variation

Cl = Confidence Interval

LP = Low Positive

MP = Medium Positive

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Image /page/9/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, the word "DiaSorin" is written in a bold, dark blue font. Below "DiaSorin", the word "Molecular" is written in a lighter green font.

510(k) Summary Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 7 of 18

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

The Limit of Detection (LoD) of the Simplexa™ COVID-19 & Flu A/B Direct assay in nasopharyngeal swabs (NPS) was determined to be the lowest detectable concentration of quantitated titered viral stocks (copies/mL or International Units/mL) at which ≥ 95% of all replicates were detected. Two (2) strains of influenza A, two (2) strains of influenza B and two (2) strains SARS-CoV-2 serially diluted in negative nasopharyngeal swab (NPS) matrix were used to determine the LoD. The LoD results are shown in Table 5.

Table 5. Simplexa™ COVID-19 & Flu A/B Direct Limit of Detection

Virus Strain	LoD (copies/mL)
Influenza A/Hong Kong/8/68	500
Influenza A/Michigan/45/2015	500
Influenza B/Phuket/3073/2013	750
Influenza B/Malaysia/2506/2004	250
SARS-CoV-2 (USA-WA1/2020)	500

ANALYTICAL REACTIVITY

Analytical Reactivity - Influenza A and B and SARS-CoV-2 (Wet-testing)

Analytical reactivity was evaluated with nasopharyngeal swab (NPS) matrix for the Simplexa™ COVID-19 & Fly AB Direct assay. A total of 63 Flu A strains and five (5) SARS-CoV-2 strains were tested. Quantified viral material was spiked into neqative NPS matrix at the concentrations listed in Tables 6-8 below and assayed in triplicate. The results are shown in Tables 6-8. Additional testing of all influenza strains in the CDC panels for 2018-2021 was performed. The results are also shown in Tables 6-7 and the CDC panels tested highlighted in Tables 9 - 11. All strains and subtypes were 100% detected with the Simplexa™ COVID-19 & Flu A/B Direct assay.

Organism	TestedConcentration*	% Detected
A/Anhui/1/2013	1:100,000 Dilution	100.0% (3/3)
A/black-legged kittiwake/Quebec/02838-1/2009	100 CEID50/mL	100.0% (3/3)
A/Brisbane/02/2018	100 EID50/mL	100.0% (3/3)
A/Brisbane/10/07	100 TCID50/mL	100.0% (3/3)
A/Brisbane/59/07	100 TCID50/mL	100.0% (3/3)
A/American green-winged teal/Mississippi/300/2010	100 CEID50/mL	100.0% (3/3)
A/California/02/2014	100 TCID50/mL	100.0% (3/3)
A/California/4/2009	100 TCID50/mL	100.0% (3/3)
A/California/7/2009	100 TCID50/mL	100.0% (3/3)
A/chicken/Germany/N/49	100 CEID50/mL	100.0% (3/3)
A/chicken/Vietnam/NCVD-016/2008(H5N1)-PR8-IDCDC-RG12	1:100,000 Dilution	100.0% (3/3)
A/Christ Church/16/2010	1,000 EID50/mL	100.0% (3/3)
A/duck/Chabarovsk/1610/1972	100 CEID50/mL	100.0% (3/3)
Organism	TestedConcentration*	% Detected
A/duck/Czechoslovakia/1956	5,000 CEID50/mL	100.0% (3/3)
A/duck/Wisconsin/480/1979	100 CEID50/mL	100.0% (3/3)
A/Egypt/N03072/2010(H5N1)-PR8-IDCDC-RG29	1:100,000 Dilution	100.0% (3/3)
A/Guangdong-Maonan/1536/2019	100 EID50/mL	100.0% (3/3)
A/Hawaii/15/2001	100 CEID50/mL	100.0% (3/3)
A/Hong Kong/2671/2019	100 EID50/mL	100.0% (3/3)
A/Hong Kong/4801/2014	100 TCID50/mL	100.0% (3/3)
A/Hong Kong/33982/2009(H9N2)-PR8-IDCDC_RG26	100 CEID50/mL	100.0% (3/3)
A/Hubei/1/2010(H5N1)-PR8-IDCDC-RG30	1:100,000 Dilution	100.0% (3/3)
A/India/NIV/2006(H5N1)-PR8-IBCDC-RG7	1:100,000 Dilution	100.0% (3/3)
A/Indiana/08/2011	100 TCID50/mL	100.0% (3/3)
A/Kansas/14/2017	100 EID50/mL	100.0% (3/3)
A/mallard/Netherlands/12/2000(H7N7)/PR8-IBCDC-1	1:100,000 Dilution	100.0% (3/3)
A/mallard/Illinois/10OS4334/2010	100 CEID50/mL	100.0% (3/3)
A/mallard/Wisconsin/4218/2009	100 CEID50/mL	100.0% (3/3)
A/mallard/Wisconsin/4230/2009	100 CEID50/mL	100.0% (3/3)
A/Massachusetts/15/2013	100 CEID50/mL	100.0% (3/3)
A/Mexico/4108/2009	100 CEID50/mL	100.0% (3/3)
A/Minnesota/11/2010	100 CEID50/mL	100.0% (3/3)
A/Minnesota/19/2011	100 CEID50/mL	100.0% (3/3)
A/New Caledonia/20/99	100 TCID50/mL	100.0% (3/3)
A/New York/18/2009	100 CEID50/mL	100.0% (3/3)
A/New York/55/2004	100 CEID50/mL	100.0% (3/3)
A/NY/02/09	100 TCID50/mL	100.0% (3/3)
A/Ohio/02/2012	100 CEID50/mL	100.0% (3/3)
A/Perth/16/2009	100 EID50/mL	100.0% (3/3)
A/pheasant/New Jersey/1355/1998(H5N2)-PR8-IBCDC-4	1:100,000 Dilution	100.0% (3/3)
A/Port Chalmers/1/1973	100 TCID50/mL	100.0% (3/3)
A/PR/8/34	100 TCID50/mL	100.0% (3/3)
A/quail/Italy/1117/1965	100 CEID50/mL	100.0% (3/3)
A/red knot/Delaware Bay/240/1994	100 CEID50/mL	100.0% (3/3)
A/red knot/Delaware/541/1988	1,000 CEID50/mL	100.0% (3/3)
A/redhead/Alberta/192/2002	100 CEID50/mL	100.0% (3/3)
A/Rhode Island/01/2010	100 CEID50/mL	100.0% (3/3)

Table 6. Simplexa™ COVID-19 & FLU A/B Direct Analytical Reactivity Results - Flu A

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Image /page/10/Picture/0 description: The image shows the text 'K220963' in a bold, sans-serif font. The text is black and appears against a white background. The characters are evenly spaced and easily readable.

Image /page/10/Picture/1 description: The image shows the logo for DiaSorin Molecular. The logo consists of a DNA strand graphic in 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 text is aligned to the right of the DNA graphic.

510(K) Summary COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 8 of 18

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Image /page/11/Picture/0 description: The image shows the alphanumeric string 'K220963' in a bold, sans-serif font. The characters are uniformly sized and spaced, creating a clear and legible sequence. The string appears to be a code or identifier, possibly a serial number or product key.

Image /page/11/Picture/1 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 text "DiaSorin" in a dark blue, sans-serif font, with "Molecular" underneath in a lighter green color. The overall design is clean and modern, suggesting a focus on biotechnology and molecular diagnostics.

510(k) Summary Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023

Page 9 of 18

Organism	TestedConcentration*	% Detected
A/Santiago/7981/2006	100 CEID50/mL	100.0% (3/3)
A/shorebird/Delaware Bay/211/1994	1,000 CEID50/mL	100.0% (3/3)
A/shorebird/Delaware/172/2006	1,000 CEID50/mL	100.0% (3/3)
A/Singapore/INFIMH-16-0019/2016	100 TCID50/mL	100.0% (3/3)
A/Solomon Island/3/2006	100 TCID50/mL	100.0% (3/3)
A/Swine/1976/31	100 TCID50/mL	100.0% (3/3)
A/Swine/lowa/15/30	100 TCID50/mL	100.0% (3/3)
A/swine/Ohio/09SW1477/2009	100 TCID50/mL	100.0% (3/3)
A/swine/Ohio/09SW83E/2009	100 CEID50/mL	100.0% (3/3)
A/Switzerland/9715293/2013	100 CEID50/mL	100.0% (3/3)
A/Taiwan/42/06	100 TCID50/mL	100.0% (3/3)
A/turkey/Massachusetts/3740/1965	2,000 CEID50/mL	100.0% (3/3)
A/turkey/Virginia/4529/2002 (H7N2)xPR8-IBCDC-5	1:100,000 Dilution	100.0% (3/3)
A/Texas/50/2012	100 TCID50/mL	100.0% (3/3)
A/Wisconsin/67/05	100 TCID50/mL	100.0% (3/3)
A/WS/33	100 TCID50/mL	100.0% (3/3)

*TCID50/mL = Tissue Culture Infectious Dose

CEID50/mL = Chicken Embryo Infectious Dose

EID50/mL = Egg Infectious Dose

Table 7. Simplexa™ COVID-19 & Flu A/B Direct Analytical Reactivity Results - Flu B

Organism	TestedConcentration*	% Detection
B/Brisbane/33/2008	100 CEID50/mL	100% (3/3)
B/Brisbane/60/2008	100 TCID50/mL	100% (3/3)
B/Christchurch/33/2004	100 TCID50/mL	100% (3/3)
B/Colorado/06/2017	100 TCID50/mL	100% (3/3)
B/Florida/02/2006	100 TCID50/mL	100% (3/3)
B/Florida/04/2006	100 TCID50/mL	100% (3/3)
B/Florida/07/04	100 TCID50/mL	100% (3/3)
B/Great Lakes/1739/54	100 TCID50/mL	100% (3/3)
B/Guangdong-Liwan/1133/2014	1,000 CEID50/mL	100% (3/3)
B/Maryland/1/59	100 TCID50/mL	100% (3/3)

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Image /page/12/Picture/0 description: The image contains the logo for DiaSorin Molecular. The logo consists of a DNA helix graphic on the left, with the text "DiaSorin" in a dark blue font to the right of the helix. Below "DiaSorin" is the word "Molecular" in a light green font.

510(k) Summary

Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 10 of 18

Organism	TestedConcentration*	% Detection
B/Massachusetts/02/2012	100 TCID50/mL	100% (3/3)
B/Michigan/09/2011	100 EID50/mL	100% (3/3)
B/Nevada/03/2011	100 CEID50/mL	100% (3/3)
B/New Hampshire/01/2016	100 EID50/mL	100% (3/3)
B/Panama/45/90	100 TCID50/mL	100% (3/3)
B/Texas/02/2013	100 TCID50/mL	100% (3/3)
B/Texas/81/2016	100 EID50/mL	100% (3/3)
B/Utah/09/2014	100 CEID50/mL	100% (3/3)
B/Victoria/304/2006	100 CEID50/mL	100% (3/3)
B/Washington/02/2019	100 EID50/mL	100% (3/3)
B/Wisconsin/01/2010	100 CEID50/mL	100% (3/3)

*TCID50/mL = Tissue Culture Infectious Dose

CEID50/mL = Chicken Embryo Infectious Dose

EID50/mL = Egg Infectious Dose

Table 8. Simplexa™ COVID-19 & Flu A/B Direct Analytical Reactivity Results - SARS-CoV-2
-----------------------------------------------------------------------------------------

Organism	TestedConcentration*	% Detection
England/204820464/2020	1000 copies/mL	100% (3/3)
hCoV19/USA/PHC658/2021	1500 copies/mL	100% (3/3)
HongKong/VM200001061/2020	1000 copies/mL	100% (3/3)
Japan/TY7-503/2021	1000 copies/mL	100% (3/3)
South Africa/KRISP-EC-K005325/2020	1000 copies/mL	100% (3/3)

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Image /page/13/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, stacked above the word "Molecular" in a lighter green, sans-serif font.

510(k) Summary Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 11 of 18

Table 9 2018-2019 CDC panel Flu A and Flu B Strains Tested with Simplexa™ COVID-19 & Flu A/B Direct

Virus	Subtype	Organism
Flu A	A(H3N2)	A/Perth/16/2009
	A(H3N2)	A/Singapore/INFIMH-16-0019/2016*
Flu A	A(H1N1) pdm09	A/California/07/2009
	A(H1N1) pdm09	A/Michigan/45/2015*
Flu B	B(Victoria lineage)	B/Brisbane/60/2008
	B(Victoria lineage)	B/Colorado/06/2017*
Flu B	B(Yamagata lineage)	B/Wisconsin/01/2010
	B(Yamagata lineage)	B/Phuket/3073/2013*

*WHO recommended vaccine strains

Table10. 2019-2020 CDC panel Flu A and Flu B Strains Tested with Simplexa™ COVID-19 & Flu A/B Direct

Virus	Subtype	Organism
Flu A	A (H3N2)	A/Perth/16/2009
	A/Kansas/14/2017*
	A (H1N1) pdm09	A/Christ Church/16/2010
		A/Brisbane/02/2018*
Flu B	B (Victoria lineage)	B/Michigan/09/2011
		B/Colorado/06/2017*
	B (Yamagata lineage)	B/New Hampshire/01/2016
		B/Phuket/3073/2013*

*WHO recommended vaccine strains

Table 11. 2020-2021 CDC panel Flu B Strains Tested with Simplexa™ COVID-19 & Flu A/B Direct

Virus	Subtype	Organism
Flu A	A(H3N2)	A/Perth/16/2009
	A/Hong Kong/2671/2019*
Flu A	A(H1N1) pdm09	A/Christ Church/16/2010
	A/Guangdong-Maonan/1536/2019*
Flu B	B(Victoria lineage)	B/Michigan/09/2011
	B/Washington/02/2019*
Flu B	B(Yamagata lineage)	B/Texas/81/2016
	B/Phuket/3073/2013*

*WHO recommended vaccine strain

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Image /page/14/Picture/0 description: The image contains 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.

510(k) Summary Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 12 of 18

Analytical Reactivity/Inclusivity - SARS-CoV-2

An in silico inclusivity analysis of the SARS-CoV-2 target primers and probes in the Simplexa™ COVID-19 & Flu AB 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 reverse primer as well as probe region were included in the analysis showed that the Simplexa™ COVID-19 & Flu A/B Direct target regions had no mismatch to 2,170,382 sequences (~99.99%) and those sequences 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 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 Mismatics 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 are not affected by the mismatches.

In addition, an analysis was conducted for the oligonucleotide (oligo) seguences for the SARS-CoV-2 ORF1ab and S gene sets against all SARS-CoV-2 sequences available in the GISAID database submitted from May 01, 2022 to July 31, 2022. In this sequence set, there were 211,184 sequences (~99.98%) with no mismatches in at least one gene oligo set and 40 (~0.02%) sequences with mismatches in at least one oligo binding region for both gene oligo sets. Based on in silico analysis of the percent homology between assay oligos and target sequences, potential impact of location of the mismatches on extension and/or binding, and the mismatch Tm values of the oligo sequence to its binding region on each analyzed SARS-CoV-2 sequence, it is predicted that the assay will also detect 100% of theses 211,224 SARS-CoV-2 sequences available in the database from May 01, 2022. A more recent in silico inclusivity analysis of the oligonycleotide (oligo) sequences for the SARS-CoV-2 ORF1ab and S gene sets was performed against all SARS-CoV-2 sequences submitted to the GISAID EpiCoV database from November 01, 2022 to February 08, 2023. The analysis as described above in previous studies, showed that the Simplexa™ COVID-19 & Flu A/B Direct target is predicted to detect the additional 12,378 sequences (100%) analyzed during this period. Table 12 below summarizes the Tm analysis results from the studies.

Table 12. Summary of Tm Analysis Results

No. seq. where at least one gene oligo set meets Tm criteria	2,394,186
No. seq. where no gene oligo set meets Tm criteria	0

Cross-Reactivity (Analytical Specificity)

Cross-reactivity of the Simplexa™ COVID-19 & Flu A/B Direct assay was evaluated by testing whole organisms or purified nucleic acid from other organisms. Specimens for laboratory testing cultured isolates/inactivated organisms/purified nucleic acids (whole genome) into negative matrix (NPS) and determining cross reactivity based on three replicates. Results from cross-reactivity testing are summarized in Table 13.

	Table 13. Simplexa™ COVID-19 & Flu A/B Direct Cross-Reactivity Results

Organism	TestedConcentration1	SARS-CoV-2	Flu A	Flu B
Adenovirus Type 1	1 x 105 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Adenovirus Type 7A	1 x 105 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Bordetella pertussis	1 x 106 CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Candida albicans	1 x 106 CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Chlamydia pneumoniae	1 x 106 IFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Organism	TestedConcentration¹	SARS-CoV-2	Flu A	Flu B
Page 1
Corynebacterium diphtheriae	1 x 10⁶ CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Coxiella burnetii	1 x 10⁶ genomecopies/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Cytomegalovirus	1 x 10⁵ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Enterovirus Type 68	1 x 10⁵ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Enterovirus Type 71	1 x 10⁵ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Epstein-Barr Virus	1 x 10⁵ copies/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Escherichia coli O157:H7	1 x 10⁶ CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Haemophilus influenzae	1 x 10⁶ CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Human Coronavirus 229E*	1 x 10⁴ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Human Coronavirus NL63*	1 x 10⁴ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Human Coronavirus OC43	1 x 10⁵ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Human Metapneumovirus 9*	1 x 10⁴ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Lactobacillus plantarum,17-5	1 x 10⁶ CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Legionella longbeachae	1 x 10⁶ CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Legionella pneumophila	1 x 10⁶ CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Leptospira interrogans	1 x 10⁶ copies/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Measles	1 x 10⁵ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
MERS-Coronavirus	1 x 10⁵ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Moraxella catarrhalis	1 x 10⁶ CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Mumps	1 x 10⁵ TCID₅₀/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Mycobacterium tuberculosis GenomicDNA	1 x 10⁶ copies/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Mycoplasma pneumoniae	1 x 10⁶ CCU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Neisseria elongata	1 x 10⁶ CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Neisseria meningitidis	1 x 10⁶ CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Organism	TestedConcentration1	SARS-CoV-2	Flu A	Flu B
Parainfluenza Type 1	1 x 105 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Parainfluenza Type 2	1 x 105 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Parainfluenza Type 3	1 x 105 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Parainfluenza Type 4	1 x 105 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Parechovirus Type 3	1 x 105 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Pseudomonas aeruginosa	1 x 106 CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Rhinovirus 1A*	1 x 104 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
RSV-A	1 x 105 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
RSV-B	1 x 105 TCID50/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Staphylococcus aureus	1 x 106 CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Staphylococcus epidermidis	1 x 106 CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Streptococcus pneumoniae	1 x 106 CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Streptococcus pyogenes	1 x 106 CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Streptococcus salivarius	1 x 106 CFU/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Human Coronavirus RNA HKU1	1 x 105 genomecopies/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Human Genomic DNA (Leukocytes)	1 x 106 cells/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
Pooled Human Nasal Wash	1:1 dilution	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)
SARS-COV1 Synthetic RNA	1 x 105 genomecopies/mL	0.0%(0/3)	0.0%(0/3)	0.0%(0/3)

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Image /page/15/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a green and blue DNA helix graphic on the left. To the right of the graphic is the text "DiaSorin" in dark blue, with the word "Molecular" underneath in green.

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510(k) Summary

Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 14 of 18

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

1CCU/mL = Color changing units/milliliter

CFU/mL= Colony forming units/milliliter

IFU/mL = Infectious units/milliliter

TCID50/mL = Tissue Culture Infectious Dose

INTERFERING SUBSTANCES

Potentially interfering substances from respiratory specimens were tested for ability to generate false negative results. Samples were prepared by spiking each potentially interfering substance into a baseline sample consisting of pooled negative nasopharyngeal swab specimens and SARS-CoV-2 inactivated viral particles (2019-nCoV/USA-WA1/2020 strain), Influenza A/Hong Kong/8/68 and Influenza B/Malaysia/2506/2004. The test samples contained each of the three

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(3) viruses at a concentration of 3X LoD. The results are shown in Table 14. Cold Eeze at a higher concentration of 2.5% (w/v) showed an invalid result in one out of the three replicates for Flu B. The replicates were retested at a lower concentration of 1.25% (w/v) and no interference was observed in any of the targets. No other substances tested in the table below showed any interference with the detection of SARS-CoV-2, influenza A or influenza B at the concentrations tested. The FluMist nasal vaccine was not tested as an interfering substance due to its unavailability at the time of this study.

Potentially InterferingSubstance	Active Ingredient	TestedConcentration*	SARS-CoV-2	Flu A	Flu B
Afrin Nasal spray	Oxymetazoline	15% (v/v)	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Antibacterial, systemic	Tobramycin	4 µg/mL	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Antibiotic, nasal ointment	Mupirocin	6.6 mg/mL	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Whole Blood	N/A	2% (v/v)	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Cold Eeze	N/A	2.5% (w/v)	100.0% (4/4)	100.0% (4/4)	100.0% (2/2)*
(Throat lozenges, Oralanesthetic and analgesic)	N/A	1.25% (w/v)	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Nasal corticosteroid(Beconase AQ)	Beclomethasone	5% (v/v)	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Nasal corticosteroid(Flonase)	Fluticasone	5% (v/v)	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Relenza Antiviral Drug	Zanamivir	3.3 mg/mL	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Tamiflu Antiviral drug	Oseltamivir	1 µM	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Zicam Nasal Gel	Luffa operculata,Galphimia glauca,histaminumhydrochloricum	5% (w/v)	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Zicam Nasal Spray(Homeopathic allergy reliefmedicine)	N/A	10% (v/v)	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)
Bovine submaxillary glandmucin, type I-S**	Purified MucinProtein	5mg/mL	100.0% (3/3)	100.0% (3/3)	100.0% (3/3)

Table 14. Simplexa" COVID-19 & Flu A/B Direct - Potentially Interfering Substances Results

*µg/mL = Micrograms/milliliter

mg/mL = Milligrams/milliliter

uM = Micromolar
v/v = Volume per Volume

w/v = Weight/Volume

*2.5% Cold Eeze Throat lozenges resulted in one out of three replicates with an EC500, invalid, for SARS-CoV-2. An EC500 result was obtained on the repeat. Therefore the concentration was lowered to 1.25% and retested.

** Mucin tested in a separate study using Influenza AlMichigan/45/2015 and Influenza B/Phuket/3073/2013)

COMPETITIVE INTERFERENCE

Competitive Interference was performed to assess the ability of the assay to detect low concentration of one (1) target analyte in the presence of high concentration of another target analyte. Samples were prepared by spiking one (1) assay target analyte at a low concentration (4X LoD) into negative nasopharyngeal swab (NPS) matrix in the presence of a high concentration (1000X LoD) of one (1) of the other two (2) assay target analytes. All the possible assay target combinations were tested. Each contrived sample was tested in triplicate. The results are shown in Table 15. All of the combinations tested showed no competitive interference for the detection of low concentrations of SARS-CoV-2, Flu A or Flu B in the presence of high concentrations of another assay target analyte.

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510(k) Summary Simplexa™ COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 16 of 18

Table 15. Simplexa™ COVID-19 & Flu A/B Direct - Competitive Interference Results

Low Positive Baseline Sample		Competitive Interferent		% Detection
Strain	Copies/mL	Strain	Copies/mL	Flu A	Flu B	SARS-CoV-2
Influenza A/HongKong/8/68	2000	Influenza B/Phuket/3073/2013	$7.5 x 10^5$	100%	100%	0.0%
		COVID-19/USA-WA1/2020	$5.0 x 10^5$	100%	0.0%	100%
Influenza B/Phuket/3073/2013	3000	Influenza A/HongKong/8/68	$5.0 x 10^5$	100%	100%	0.0%
		COVID-19/USA-WA1/2020	$5.0 x 10^5$	0.0%	100%	100%
COVID-19/USA-WA1/2020	2000	Influenza A/HongKong/8/68	$5.0 x 10^5$	100%	0.0%	100%
		Influenza B/Phuket/3073/2013	$7.5 x 10^5$	0.0%	100%	100%

INTERFERENCE BY OTHER MICROORGANISMS

The Simplexa™ COVID-19 & Flu A/B Direct assay was evaluated by testing the ability to identify SARS-CoV-2, influenza A virus, and influenza B virus, when other potentially interfering organisms were prepared by spiking cultured isolated organisms/purified nucleic acids (whole genome) at a minimum of 10° CFU/mL (or higher) for bacteria, and 105 TC/Ds0/mL or PFU/mL (or higher) for viruses into negative nasopharyngeal swab (NPS) matrix in the presence of a low concentration (2X LoD) of the three (3) targets (SARS-CoV-2, Flu A and Flu B viral particles) and determining microbial interference based on three (3) replicates. For organisms not titered in CFU/mL or TCIDso/mL, other industry acceptable units were used as indicated. The panel of forty seven (47) potentially inhibitory organisms was individually spiked into a pool with a low concentration influenza AlHong Kong/8/68), influenza B (Influenza BlMalaysia/2506/2004), and SARS-CoV-2 (2019-nCoV/USA-WA1/2020). No interference by other orqanisms was observed for SARS-CoV-2, influenza B at the concentrations indicated in Table 16.

Table 16. Simplexa™ COVID-19 & Flu A/B Direct – Microbial Interference Results

Organism	TestedConcentration1	% Detection
		SARS-CoV-2	Flu A	Flu B
Adenovirus Type 1	1 x 105 TCID50/mL	100%	100%	100%
Adenovirus Type 7A	1 x 105 TCID50/mL	100%	100%	100%
Bordetella pertussis	1 x 106 CFU/mL	100%	100%	100%
Candida albicans	1 x 106 CFU/mL	100%	100%	100%
Chlamydia pneumoniae	1 x 106 IFU/mL	100%	100%	100%
Corynebacterium diphtheriae	1 x 106 CFU/mL	100%	100%	100%
Coxiella burnetii	1 x 106genome copies/mL	100%	100%	100%
Cytomegalovirus	1 x 105 TCID50/mL	100%	100%	100%
Enterovirus Type 68	1 x 105 TCID50/mL	100%	100%	100%
Organism	TestedConcentration¹	% Detection
		SARS-CoV-2	Flu A	Flu B
Enterovirus Type 71	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
Epstein-Barr Virus	1 x 10⁵ copies/mL	100%	100%	100%
Escherichia coli O157:H7	1 x 10⁶ CFU/mL	100%	95%	100%
Haemophilus influenzae	1 x 10⁶ CFU/mL	100%	100%	100%
Human Coronavirus 229E*	1 x 10⁴ TCID₅₀/mL	100%	100%	100%
Human Coronavirus NL63*	1 x 10⁴ TCID₅₀/mL	100%	100%	100%
Human Coronavirus OC43	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
Human Metapneumovirus 9*	1 x 10⁴ TCID₅₀/mL	100%	100%	100%
Lactobacillus plantarum,17-5	1 x 10⁶ CFU/mL	100%	100%	100%
Legionella longbeachae	1 x 10⁶ CFU/mL	100%	100%	100%
Legionella pneumophila	1 x 10⁶ CFU/mL	100%	100%	100%
Leptospira interrogans	1 x 10⁶ copies/mL	100%	100%	100%
Measles	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
MERS-Coronavirus	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
Moraxella catarrhalis	1 x 10⁶ CFU/mL	100%	100%	100%
Mumps	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
Mycobacterium tuberculosis GenomicDNA	1 x 10⁶ copies/mL	100%	100%	100%
Mycoplasma pneumoniae	1 x 10⁶ CCU/mL	100%	100%	100%
Neisseria elongata	1 x 10⁶ CFU/mL	100%	100%	100%
Neisseria meningitidis	1 x 10⁶ CFU/mL	100%	100%	100%
Parainfluenza Type 1	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
Parainfluenza Type 2	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
Parainfluenza Type 3	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
Parainfluenza Type 4	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
Parechovirus Type 3	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
Pseudomonas aeruginosa	1 x 10⁶ CFU/mL	100%	100%	100%
Rhinovirus 1A*	1 x 10⁴ TCID₅₀/mL	95%	100%	100%
RSV-A	1 x 10⁵ TCID₅₀/mL	100%	100%	100%
RSV-B	1 x 10⁵ TCID₅₀/mL	100%	100%	100%

	Tested	% Detection
Organism	Concentration¹	SARS-CoV-2	Flu A	Flu B
Staphylococcus aureus	1 x 106 CFU/mL	100%	100%	100%
Staphylococcus epidermidis	1 x 106 CFU/mL	100%	100%	100%
Streptococcus pneumoniae	1 x 106 CFU/mL	100%	100%	100%
Streptococcus pyogenes	1 x 106 CFU/mL	100%	100%	100%
Streptococcus salivarius	1 x 106 CFU/mL	100%	100%	100%
Human Coronavirus RNA HKU1	1 x 105 genomecopies/mL	100%	100%	100%
Human Genomic DNA (Leukocytes)	1 x 106 cells/mL	100%	100%	100%
Pooled Human Nasal Wash	1:1 dilution	100%	100%	100%
SARS-COV1 Synthetic RNA	1 x 105 genomecopies/mL	100%	100%	100%

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510(K) Summary COVID-19 & Flu A/B Direct MOL4250 March 16, 2023 Page 17 of 18

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510(k) S marv

Simplexa™ COVID-19 & Flu A/B D 4250 2023 Page 18 of 18

"A lower concentration was tested due to inability to obtain stock material with high titer 1CCU/mL = Color Changing Units/milliliter

CFU/mL = Colony Forming Units/milliliter

IFU/mL = Infectious units/milliliter

TCIDso/mL = Tissue Culture Infectious Dose/milliliter

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 & Flu A/B Direct is substantially Equivalent to the predicate device (DEN200031). The device labeling is compliant with 21 CFR § 809.10.

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.