Simplexa VZV Direct, Simplexa VZV Positive Control Pack by DiaSorin Molecular LLC

The DiaSorin Molecular Simplexa™ VZV Direct assay is intended for use on the LIAISON® MDX instrument for the qualitative detection of varicella-zoster virus (VZV) DNA in cerebrospinal fluid (CSF) from patients signs and/or symptoms of meningitis and/or encephalitis. This test is intended as an aid in the diagnosis of VZV infections of the central nervous system (CNS).

Negative results do not preclude VZV infection and should not be used as the sole basis for treatment or other patient management decisions.

The assay is not intended for use as a donor screening test. The assay is for professional use only.

The Simplexa™ VZV Positive Control Pack is intended to be used as a control with the Simplexa™ VZV Direct kit.

This control is not intended for use with other assays or systems.

Device Description

The Simplexa™ VZV Direct assay is a real-time polymerase chain reaction (PCR) system that enables the direct amplification and detection of VZV DNA from unprocessed cerebral spinal fluid (CSF) specimens without nucleic acid extraction. The system consists of the Simplexa™ VZV Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc and associated accessories. In the Simplexa™ VZV Direct assay, fluorescent probes are used together with corresponding forward and reverse primers to amplify VZV and internal control targets. A well-conserved region of the VZV DNA polymerase gene is targeted to identify VZV DNA in the specimen. An internal control is used to detect PCR failure and/or inhibition.

AI/ML Overview

Here's an analysis of the acceptance criteria and the study proving the device meets them, based on the provided text:

Acceptance Criteria and Device Performance for Simplexa™ VZV Direct Assay

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly list "acceptance criteria" as a set of predefined thresholds. Instead, it presents performance metrics from various studies. Based on the provided clinical agreement study, the implicit acceptance criteria for clinical performance would likely be a high Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA). For analytical performance, the LoD is a key metric.

Metric (Implicit Acceptance Criteria)	Reported Device Performance (Simplexa™ VZV Direct)
Clinical Performance
Positive Percent Agreement (PPA)	100.0% (12/12) (95% CI: 75.7% to 100.0%)
Negative Percent Agreement (NPA)	99.7% (623/625) (95% CI: 98.8% to 99.9%)
PPA (Contrived Samples)	100.0% (120/120) (95% CI: 96.9-100.0%)
Analytical Performance
Limit of Detection (LoD) VZV Strain 9939	2.03 TCID50/mL (1,614 copies/mL)
Limit of Detection (LoD) VZV Strain Ellen	0.001 TCID50/mL (1,505 copies/mL)
Analytical Reactivity (VZV strains)	100.0% agreement for 5 additional VZV strains
Cross-Reactivity (159 microorganisms)	0.0% detection (no cross-reactivity observed)
Reproducibility (VZV) %CV	0.5-4.6%
Interference	No interference observed
Inhibition by other microorganisms	No inhibition observed

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

Sample Size (Clinical):
- Prospective Samples: 637 clinical samples.
- Contrived Samples: 240 contrived VZV positive samples.
Data Provenance:
- Country of Origin: Not explicitly stated, but the submission is to the U.S. FDA by a company in Cypress, California, USA, and the testing was performed at "testing sites" and "DiaSorin Molecular, Cypress, CA", suggesting U.S.-based data.
- Retrospective or Prospective: The study included both:
  - Prospective and prospectively banked collections from eight (8) collection sites (February 2018 to November 2018).
  - Contrived samples were used to supplement low prevalence true positive clinical samples.

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

Not applicable. This device is an in vitro diagnostic (IVD) based on molecular detection. Ground truth for diagnostic assays like this is typically established by comparative methods rather than expert human interpretation of images or other subjective data. No human "experts" were used to establish the ground truth in the sense of clinical decision-making from images.

4. Adjudication Method for the Test Set

Not applicable in the sense of expert human review (e.g., 2+1, 3+1). The ground truth for the clinical samples was established using a composite reference method:

Two (2) validated real-time PCR assays.
Followed by confirmation of positive PCR amplification products with bi-directional sequencing.
Decision Rule: Samples were characterized as positive if one (1) or both PCR assays were positive AND confirmed by bi-directional sequencing. Samples were characterized as negative if both PCR assays were negative.

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

No, an MRMC comparative effectiveness study was not done. This device is an in vitro diagnostic (IVD) designed for laboratory use to detect VZV DNA. Its performance is evaluated against reference methods, not against human readers (e.g., radiologists, pathologists). Therefore, improvement for human readers with AI assistance is not applicable in this context.

6. Standalone (Algorithm Only Without Human-in-the-Loop Performance)

Yes, the primary performance studies presented focus on the standalone performance of the Simplexa™ VZV Direct assay. The "Clinical Agreement" section directly compares the Simplexa™ VZV Direct results to the composite reference method without explicit human intervention in the interpretation of the Simplexa™ VZV Direct results. The assay is an automated real-time PCR system.

7. Type of Ground Truth Used

The ground truth for the clinical agreement study was established using a composite reference method. This method involved:

Two (2) validated real-time PCR assays.
Confirmation of positive PCR amplification products with bi-directional sequencing.

8. Sample Size for the Training Set

The document does not explicitly mention a "training set" in the context of machine learning or AI models. This device is an in vitro diagnostic (IVD) based on real-time PCR technology, which relies on molecular biology principles (primers and probes) rather than machine learning models that require distinct training and test sets. The assay design and optimization would involve internal development and validation, but not typically a "training set" in the AI sense.

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

Not applicable, as a "training set" for an AI model is not described or relevant for this type of molecular diagnostic device. The analytical characteristics and design of the PCR assay (e.g., primer and probe specificity) are established through various analytical studies (e.g., analytical sensitivity, specificity, cross-reactivity) rather than a ground-truthed training set for machine learning.

Summary

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May 13, 2019

DiaSorin Molecular LLC Sharon Young Principal Regulatory Affairs Specialist 11331 Valley View Street Cypress, California 90630

Re: K190219

Trade/Device Name: Simplexa VZV Direct, Simplexa VZV Positive Control Pack Regulation Number: 21 CFR 866.3970 Regulation Name: Device to detect and identify microbial pathogen nucleic acids in cerebrospinal fluid Regulatory Class: Class II Product Code: PLO Dated: January 30, 2019 Received: February 4, 2019

Dear Sharon Young:

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 statutes and regulations administered by other Federal agencies. You must comply with all the Act's

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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/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (OS) 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 http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). 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 (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely yours.

for

Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K190219

Device Name

Simplexa VZV Direct, Simplexa VZV Positive Control Pack

Indications for Use (Describe)

Negative results do not preclude VZV infection and should not be used as the sole basis for treatment or other patient management decisions.

The assay is not intended for use as a donor screening test. The assay is for professional use only.

Type of Use (Select one or both, as applicable)X Prescription Use (Part 21 CFR 801 Subpart D)

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

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510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 1 of 25

Applicant	DiaSorin Molecular LLC.11331 Valley View StreetCypress, California 90630USA
Establishment Registration No.	2023365
Contact Person	Sharon YoungPrincipal Regulatory Affairs Specialisttel 562.240.6680fax 562.240.6529Sharon.Young@DiaSorin.com
Summary Date	May 3, 2019
Proprietary Name	Simplexa™ VZV Direct and Simplexa™ VZV Positive Control Pack
US Product Codes/Names andRegulation Numbers	PLO / Device to detect and identify microbial pathogen nucleic acidsin cerebrospinal fluid / 21 CFR 866.3970OOI / Real Time Nucleic Acid Amplification System / 21 CFR 862.2570
Classification	Class II

Intended Use

Simplexa™ VZV Direct REF MOL3650

The DiaSorin Molecular Simplexa™ VZV Direct assay is intended for use on the LIAISON® MDX instrument for the qualitative detection of varicella-zoster virus (VZV) DNA in cerebrospinal fluid (CSF) from patients with signs and/or symptoms of meningitis and/or encephalitis. This test is intended as an aid in the diagnosis of VZV infections of the central nervous system (CNS).

Neqative results do not prection and should not be used as the sole basis for treatment or other patient management decisions.

The assay is not intended for use as a donor screening test. The assay is for professional use only.

Simplexa™ VZV Positive Control Pack REF MOL3660

The Simplexa™ VZV Positive Control Pack is intended to be used as a control with the Simplexa™ VZV Direct kit.

This control is not intended for use with other assays or systems.

Device Description

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510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 2 of 25

Simplexa™ VZV Direct REF MOL3650

Component Name	REF	ECSYMBOLONLABEL		AbbreviatedName	CapColor	Numberof Vials	ReactionsperVial/Kit	Volumeper Vial
Simplexa™ VZVDirect Reaction Mix	MOL3651	REAG	C	RM	Purple	24	1/24	50 μL

Simplexa™ VZV Direct Components and Descriptions

Kit Component	Contents
Simplexa™ VZV Direct Reaction Mix (RM)	DNA polymerase, buffer, dNTPs, template DNA (Internal Control), dye-labeled fluorescent probes and primers specific for detection of VZV and for the DNA Internal Control.Target Probe Fluorophore (Dye) Excitation (nm) Emission (nm) Targeted Gene VZV FAM 495 520 VZV DNA polymerase Internal Control DNA Q670 644 670 DNA (IC)
Simplexa™ VZV Direct Kit Barcode Card	Assay specific parameters and lot information.

Simplexa™ VZV Positive Control Pack REF MOL3660 Component and Description

Component Name	REF	Description	CapColor	Numberof Vials	ReactionsperVial/Kit	Volumeper Vial
Simplexa™ VZV DirectPositive Control	MOL3661	Inactivatedvaricella zostervirus	Red	10	1/10	50 μL

Materials Supplied Separately

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

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510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 3 of 25

Comparison to Predicate Device
Comparison toPredicate Device	Predicate Device:BioFire® FilmArray®Meningitis/Encephalitis (ME) PanelK160462	Candidate Device:Simplexa™ VZV Direct and Simplexa™VZV Positive Control Pack
Product Code	PLO	Same
RegulationNumber	21 CFR 866.3970 – Device to detectand identify microbial pathogen nucleicacids in cerebrospinal fluid.	Same
OrganismDetected	varicella zoster virus	Same
Measurand	DNA from varicella zoster virus	Same
Intended Use	The FilmArray Meningitis/Encephalitis(ME) Panel is a qualitative multiplexednucleic acid-based in vitro diagnostictest intended for use with FilmArrayand FilmArray 2.0 systems. TheFilmArray ME Panel is capable ofsimultaneous detection andidentification of multiple bacterial, viral,and yeast nucleic acids directly fromcerebrospinal fluid (CSF) specimensobtained via lumbar puncture fromindividuals with signs and/or symptomsof meningitis and/or encephalitis. Thefollowing organisms are identified usingthe FilmArray ME Panel: Bacteria:Escherichia coli K1 Haemophilusinfluenzae Listeria monocytogenesNeisseria meningitidis (encapsulated)Streptococcus agalactiaeStreptococcus pneumoniae Viruses:Cytomegalovirus Enterovirus Herpessimplex virus 1 Herpes simplex virus 2Human herpesvirus 6 Humanparechovirus Varicella zoster virusYeast: Cryptococcus neoformans/gattiiThe FilmArray ME Panel is indicated asan aid in the diagnosis of specificagents of meningitis and/orencephalitis and results are meant tobe used in conjunction with otherclinical, epidemiological, and laboratorydata. Results from the FilmArray MEPanel are not intended to be used asthe sole basis for diagnosis, treatment,	Simplexa™ VZV Direct REF MOL3650The DiaSorin Molecular Simplexa™ VZVDirect assay is intended for use on theLIAISON® MDX instrument for thequalitative detection of varicella-zostervirus (VZV) DNA in cerebrospinal fluid(CSF) from patients with signs and/orsymptoms of meningitis and/orencephalitis. This test is intended as anaid in the diagnosis of VZV infections ofthe central nervous system (CNS).Negative results do not preclude VZVinfection and should not be used as thesole basis for treatment or other patientmanagement decisions.The assay is not intended for use as adonor screening test. The assay is forprofessional use only.Simplexa™ VZV Positive Control PackREF MOL3660The Simplexa™ VZV Positive ControlPack is intended to be used as a controlwith the Simplexa™ VZV Direct kit.This control is not intended for use withother assays or systems.
Comparison toPredicate Device	Predicate Device:BioFire® FilmArray®Meningitis/Encephalitis (ME) PanelK160462	Candidate Device:Simplexa™ VZV Direct and Simplexa™VZV Positive Control Pack
	decisions. Positive results do not ruleout co-infection with organisms notincluded in the FilmArray ME Panel.The agent detected may not be thedefinite cause of the disease. Negativeresults do not preclude central nervoussystem (CNS) infection. Not all agentsof CNS infection are detected by thistest and sensitivity in clinical use maydiffer from that described in thepackage insert. The FilmArray MEPanel is not intended for testing ofspecimens collected from indwellingCNS medical devices. The FilmArrayME Panel is intended to be used inconjunction with standard of careculture for organism recovery,serotyping, and antimicrobialsusceptibility testing.
AutomatedSystem (Sampleto Answer)	Yes	Yes
Instrumentation	FilmArray or FilmArray 2.0 system	LIAISON® MDX

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510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 4 of 25

CLINICAL AGREEMENT

A total of six hundred thirty-seven (637) clinical samples, meeting inclusion criteria were obtained from prospective and prospectively banked collections from eight (8) collection sites from February 2018 to November 2018. The study was conducted using remnant cerebrospinal fluid (CSF) samples from human patients suspected of VZV infection of the central nervous system (CNS).

Due to the low prevalence of VZV infection in CSF samples, two hundred forty (240) contrived samples were included to supplement the number of positive samples were contrived across the clinical range of the Simplexa™ VZV Direct assay and thirty percent (30%) of the samples were contrived at approximately 2X LoD. The samples were stored at 2-8 °C for up to seven (7) days post collection and at <-70 ℃ thereafter. All Simplexa™ VZV Direct testing was performed at the testing sites.

Simplexa™ VZV Direct results were compared to results from a composite reference method. The composite reference method consisted of two (2) validated real-time PCR assays followed by confirmation of positive PCR amplification products with bi-directional sequencing. Samples were characterized as positive if one (1) or both PCR assays were positive and confirmed by bi-directional sequencing. Samples were characterized as negative if both PCR assays were negative. Samples were tested on Simplexa™ VZV Direct at the collection sites and the composite reference method was performed at DiaSorin Molecular, Cypress, CA.

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510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 5 of 25

Table 1 shows the Clinical Agreement of the Simplexa™ VZV Direct as compared to the composite reference method of two (2) validated real-time PCR assays followed by confirmation of positive PCR amplification products with bi-directional sequencing.

Table 1. Prospective Clinical Agreement Results for Simplexa™ VZV Direct vs Composite
Reference Method

Simplexa™ VZV Direct	Composite PCR/Sequencing
Results	Detected	Not Detected	Total
Detected	12	2a	14
Not Detected	0	623	623
Total	12	625	637
	PPA100.0% (12/12)95% CI: 75.7% to100.0%	NPA99.7% (623/625)95% CI: 98.8% to99.9%

3 One (1) of two (2) discordant results were positive with an alternate NAAT used by collection site during routine testing.

Due to the low prevalence of VZV infection in CSF samples, contrived positive samples were tested to supplement the number of positive samples tested. For each of the two strains, VZV 9939 60 samples were contrived across the clinical range of the Simplexa VZV Direct assay with 60% of the samples contrived as low positive samples close to the LoD of the test. Samples were tested at four clinical sites in a blinded manner, randomized with VZV negative samples were stored at 2-8 °C for up to 7 days post collection and at <-70 °C thereafter. Simplexa VZV Direct test results for contrived samples were compared to the same Composite Reference Method described for the prospective clinical study above. The contrived sample study demonstrated a positive percent agreement of 100% (120/120) with a 95% Confidence Interval of 96.9-100.0%.

REPRODUCIBILITY

Reproducibility for the Simplexa™ VZV Direct assay was evaluated. Three (3) investigative sites assessed the device's inter-site, inter-day and inter/intra-assay reproducibility. Each of the laboratories tested Simplexa™ VZV Direct Positive Control, No Template Control (Synthetic CSF), and four (4) contrived samples in negative matrix. Two (2) strains of VZV were used in the study, 9939 and Ellen. The four (4) contrived samples consisted of a low positive (LP) and medium positive (MP) for each VZV strain. The assays were performed in triplicate on five (5) different days. Each site had two (2) operators who each assayed the entire sample panel and Positive Control once per day, for a total of two (2) sets of data per day on a total of six (6) LIAISON® MDX instruments. The combined results for all sites are presented in Tables 2 and 3. The results show the reproducibility of the Simplexa™ VZV Direct % CV to range between 0.5-4.6.

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510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 6 of 25

	Site 1		Site 2		Site 3		Total %AgreementwithExpectedResults	95% CI
SampleVZV Strain(FAM)	%AgreementwithExpectedResults	Avg.Ct	Total%CV	%AgreementwithExpectedResults	Avg.Ct	Total%CV	%AgreementwithExpectedResults	Avg.Ct	Total%CV
9939 -LP	100.0%(30/30)	35.8	4.6	100.0%(30/30)	35.7	3.6	100.0%(30/30)	36.1	4.2	100.0%(90/90)	95.9% to100.0%
9939 -MP	100.0%(30/30)	35.3	2.2	100.0%(30/30)	34.6	2.4	100.0%(30/30)	35.0	2.3	100.0%(90/90)	95.9% to100.0%
Ellen -LP	100.0%(30/30)	36.7	2.6	100.0%(30/30)	36.9	2.2	100.0%(30/30)	36.9	1.6	100.0%(90/90)	95.9% to100.0%
Ellen -MP	100.0%(30/30)	35.6	1.6	100.0%(30/30)	35.6	1.0	100.0%(30/30)	35.6	1.1	100.0%(90/90)	95.9% to100.0%
PositiveControl	100.0%(30/30)	30.3	1.1	100.0%(30/30)	29.8	1.2	100.0%(30/30)	30.7	0.9	100.0%(90/90)	95.9% to100.0%
NTC	100.0%(30/30)	0.0	N/A	100.0%(30/30)	0.0	N/A	100.0%(30/30)	0.0	N/A	100.0%(90/90)	95.9% to100.0%
TotalAgreement		100.0% (180/180)95% CI: 97.9% to 100.0%		100.0% (180/180)95% CI: 97.9% to 100.0%		100.0% (180/180)95% CI: 97.9% to 100.0%		100.0% (540/540)95% CI: 99.3% to100.0%

Table 2. Simplexa™ VZV Direct Reproducibility - VZV (FAM)

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

The Limit of Detection (LoD) was determined for the Simplexa™ VZV Direct assay using quantified stocks of two (2) VZV strains (Ellen and 9939) serially diluted into negative human CSF matrix. The LoD was determined to be the lowest concentration that could be detected positive > 95% of the time.

	Table 3. Simplexa™ VZV Direct Limit of Detection
--	--------------------------------------------------	--	--	--	--	--

VZV strain	Concentration
	TCID50/mL	copies/mL
9939	2.03 TCID50/mL	1,614
Ellen	0.001 TCID50/mL	1,505

ANALYTICAL REACTIVITY/CROSS REACTIVITY

Analytical Reactivity

The analytical reactivity of the Simplexa™ VZV Direct assay was evaluated using different strains of VZV that were not used in the determination of the limit of detection (LoD) for the assay. Quantified viral material was spiked into negative CSF using a single dilution and assayed in triplicate. The Simplexa™ VZV Direct assay was able to detect other strains of VZV at 2X LoD. The results are presented in Table 4. In addition, in silico BLAST analysis indicated that the assay should detect at least one hundred seventy-eight (178) additional VZV strains.

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510(k) Summary Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 7 of 25

VZV Strain	Agreement withExpected Results(#Detected/#Total)
VZV Strain 82	3/3
VZV Strain 275	3/3
VZV Strain 1700	3/3
VZV Isolate A	3/3
VZV Isolate B	3/3

Table 4. Simplexa™ VZV Direct Analytical Reactivity

Cross-Reactivity (Analytical Specificity)

The Simplexa™ VZV Direct assay's analytical specificity was evaluated by testing the ability of the assay to exclusively identify VZV virus with no cross-reactivity to organisms that are closely related, or cause similar clinical symptoms or may be present in CSF. One hundred and fifty-nine (159) microorganisms were spiked into negative CSF and assayed in triplicate. Some organisms with low concentrations (<1 x 10° CFU/mL for bacteria, fungi, cells or parasites; <1 x 105 IU/mL or PFU/mL or TCIDs/mL for viruses) were additionally tested in silico. For organisms not available to be tested, in silico analysis was performed. No cross-reactivity was observed. The results are presented in Table 5.

	Table 5. Simplexa™ VZV Direct Cross-Reactivity (Analytical Specificity)

No.	Organism	Tested Concentration	Agreement with ExpectedResults: % Detection(# Detected/#Tested)
1	Adenovirus A12**	NA	NA
2	Adenovirus B35**	NA	NA
3	Adenovirus B7A	1 x 105 IU/mL	0.0% (0/3)
4	Adenovirus C1	1 x 105 IU/mL	0.0% (0/3)
5	Adenovirus C2	1 x 105 IU/mL	0.0% (0/3)
6	Adenovirus D20	1 x 105 IU/mL	0.0% (0/3)
7	Adenovirus E4**	NA	NA
8	Adenovirus F41**	NA	NA
9	Aspergillus fumigatus	1 x 106 CFU/mL	0.0% (0/3)
10	Bacillus cereus	1 x 106 CFU/mL	0.0% (0/3)
11	Bacillus subtilis	1 x 106 CFU/mL	0.0% (0/3)
12	BK virus	1 x 105 IU/mL	0.0% (0/3)
13	Candida albicans	1 x 106 CFU/mL	0.0% (0/3)
No.	Organism	Tested Concentration	Agreement with ExpectedResults: % Detection(# Detected/#Tested)
14	Candida krusei	1 x 106 CFU/mL	0.0% (0/3)
15	Candida parapsilosis	1 x 106 CFU/mL	0.0% (0/3)
16	Candida tropicalis	1 x 106 CFU/mL	0.0% (0/3)
17	Citrobacter freundii	1 x 106 CFU/mL	0.0% (0/3)
18	Citrobacter koseri	1 x 106 CFU/mL	0.0% (0/3)
19	Coronavirus 229E*	1 x 104 IU/mL*	0.0% (0/3)
20	Coronavirus NL63*	1 x 104 IU/mL*	0.0% (0/3)
21	Coronavirus OC43	1 x 105 TCID50/mL	0.0% (0/3)
22	Corynebacterium striatum**	NA	NA
23	Corynebacteriumurealyticum**	NA	NA
24	Coxsackievirus A10**	NA	NA
25	Coxsackievirus A16	1 x 105 TCID50/mL	0.0% (0/3)
26	Coxsackievirus A17**	NA	NA
27	Coxsackievirus A21*	1 x 104 TCID50/mL*	0.0% (0/3)
28	Coxsackievirus A24**	NA	NA
29	Coxsackievirus A6**	NA	NA
30	Coxsackievirus A9	1 x 105 TCID50/mL	0.0% (0/3)
31	Coxsackievirus B1	1 x 105 IU/mL	0.0% (0/3)
32	Coxsackievirus B2	1 x 105 IU/mL	0.0% (0/3)
33	Coxsackievirus B3	1 x 105 TCID50/mL	0.0% (0/3)
34	Coxsackievirus B4	1 x 105 TCID50/mL	0.0% (0/3)
35	Coxsackievirus B5*	1 x 104 TCID50/mL*	0.0% (0/3)
36	Cronobacter sakazakii	1 x 106 CFU/mL	0.0% (0/3)
37	Cryptococcus albidus**	NA	NA
38	Cryptococcus amylolentus**	NA	NA
39	Cryptococcus gattii**	NA	NA
40	Cryptococcus laurentii**	NA	NA
No.	Organism	Tested Concentration	Agreement with ExpectedResults: % Detection(# Detected/#Tested)
41	Cryptococcus neoformans	1 x 106 CFU/mL	0.0% (0/3)
42	Cryptococcus uniguttulatus**	NA	NA
43	Cytomegalovirus (AD169Strain)	1 x 105 IU/mL	0.0% (0/3)
44	Dengue virus (Type 1)*	1 x 104 IU/mL*	0.0% (0/3)
45	Dengue virus (Type 2)	1 x 105 IU/mL	0.0% (0/3)
46	Echovirus 1	1 x 105 IU/mL	0.0% (0/3)
47	Echovirus 11	1 x 105 IU/mL	0.0% (0/3)
48	Echovirus 18**	NA	NA
49	Echovirus 4	1 x 105 IU/mL	0.0% (0/3)
50	Echovirus 6	1 x 105 IU/mL	0.0% (0/3)
51	Echovirus 7	1 x 105 IU/mL	0.0% (0/3)
52	Echovirus 9	1 x 105 TCID50/mL	0.0% (0/3)
53	Encephalomyocarditis virus	1 x 105 TCID50/mL	0.0% (0/3)
54	Enterobacter aerogenes	1 x 106 CFU/mL	0.0% (0/3)
55	Enterobacter cloacae	1 x 106 CFU/mL	0.0% (0/3)
56	Enterovirus 68**	NA	NA
57	Enterovirus 70	1 x 105 IU/mL	0.0% (0/3)
58	Enterovirus 71	1 x 105 IU/mL	0.0% (0/3)
59	Epstein-Barr virus (B95-8Strain)	1 x 105 copies/mL	0.0% (0/3)
60	Escherichia coli (O157:H7)	1 x 106 CFU/mL	0.0% (0/3)
61	Escherichia coli K1**	NA	NA
62	Escherichia fergusonii**	NA	NA
63	Escherichia hermanii**	NA	NA
64	Escherichia vulneris**	NA	NA
65	Filobasidium capsuligenum**	NA	NA
66	Haemophilus ducreyi	1 x 106 CFU/mL	0.0% (0/3)
67	Haemophilus haemolyticus**	NA	NA
No.	Organism	Tested Concentration	Agreement with ExpectedResults: % Detection(# Detected/#Tested)
68	Haemophilus influenza TypeB	1 x 106 CFU/mL	0.0% (0/3)
69	Haemophilus influenzae TypeA	1 x 106 CFU/mL	0.0% (0/3)
70	Haemophilusparahaemolyticus**	NA	NA
71	Haemophilus parainfluenzae	1 x 106 CFU/mL	0.0% (0/3)
72	Hepatitis A virus	1 x 105 IU/mL	0.0% (0/3)
73	Hepatitis B virus	1 x 105 IU/mL	0.0% (0/3)
74	Hepatitis C virus	1 x 105 IU/mL	0.0% (0/3)
75	Hepatitis D virus**	NA	NA
76	HHV-6A	1 x 105 copies/mL	0.0% (0/3)
77	HHV-6B	1 x 105 copies/mL	0.0% (0/3)
78	HHV-7SB	1 x 105 IU/mL	0.0% (0/3)
79	HHV-8	1 x 105 copies/mL	0.0% (0/3)
80	HIV-1 IIIB	1 x 105 IU/mL	0.0% (0/3)
81	HIV-2 NIHZ	1 x 105 IU/mL	0.0% (0/3)
82	HPeV-1**	NA	NA
83	HPeV-2**	NA	NA
84	HPeV-3	1 x 105 IU/mL	0.0% (0/3)
85	HPeV-4**	NA	NA
86	HPeV-5**	NA	NA
87	HPeV-6**	NA	NA
88	HSV-1 (MacIntyre)	1 x 105 TCID50/mL	0.0% (0/3)
89	HSV-2 (G)	1 x 105 IU/mL	0.0% (0/3)
90	Human Rhinovirus A16	1 x 105 IU/mL	0.0% (0/3)
91	Human Rhinovirus B3**	NA	NA
92	Human Rhinovirus B83**	NA	NA
93	Influenza A/California/7/2009	1 x 105 IU/mL	0.0% (0/3)
94	Influenza A/H1N1	1 x 105 TCID50/mL	0.0% (0/3)
No.	Organism	Tested Concentration	Agreement with ExpectedResults: % Detection(# Detected/#Tested)
95	Influenza B/Florida/02/2006	1 x 105 IU/mL	0.0% (0/3)
96	JC virus (MAD-4 strain)	1 x 105 IU/mL	0.0% (0/3)
97	Klebsiella pneumoniae	1 x 106 CFU/mL	0.0% (0/3)
98	La Crosse encephalitis virus	1 x 105 TCID50/mL	0.0% (0/3)
99	Listeria innocua**	NA	NA
100	Listeria ivanovii**	NA	NA
101	Listeria monocytogenes	1 x 106 CFU/mL	0.0% (0/3)
102	Measles virus	1 x 105 IU/mL	0.0% (0/3)
103	Morganella morganii**	NA	NA
104	Mumps virus	1 x 105 IU/mL	0.0% (0/3)
105	Mycobacterium tuberculosisgenomic DNA	1.6 x 106 copies/mL	0.0% (0/3)
106	Naegleria fowleri*	1.8 x 105 cells/mL*	0.0% (0/3)
107	Neisseria gonorrhoeae	1 x 106 CFU/mL	0.0% (0/3)
108	Neisseria lactamica**	NA	NA
109	Neisseria meningitidis(serogroup A)	1 x 106 CFU/mL	0.0% (0/3)
110	Neisseria meningitidis(unencapsulated)**	NA	NA
111	Neisseria mucosa	1 x 106 CFU/mL	0.0% (0/3)
112	Neisseria sicca**	NA	NA
113	Pantoea agglomerans**	NA	NA
114	Parainfluenza Type 1	1 x 105 IU/mL	0.0% (0/3)
115	Parainfluenza Type 2	1 x 105 IU/mL	0.0% (0/3)
116	Parainfluenza Type 3	1 x 105 IU/mL	0.0% (0/3)
117	Parainfluenza Type 4	1 x 105 TCID50/mL	0.0% (0/3)
118	Parvovirus B19	1 x 105 IU/mL	0.0% (0/3)
119	Poliovirus (Type 3)	1 x 105 TCID50/mL	0.0% (0/3)
120	Propionibacterium acnes	1 x 106 CFU/mL	0.0% (0/3)
121	Proteus mirabilis Z050	1 x 106 CFU/mL	0.0% (0/3)
No.	Organism	Tested Concentration	Agreement with ExpectedResults: % Detection(# Detected/#Tested)
122	Pseudomonas aeruginosa	1 x 106 CFU/mL	0.0% (0/3)
123	Rabies virus	1 x 105 TCID50/mL	0.0% (0/3)
124	Respiratory syncytial virus	1 x 105 IU/mL	0.0% (0/3)
125	Rhinovirus 1A	1 x 105 IU/mL	0.0% (0/3)
126	Rotavirus (Type Wa)*	1 x 104 IU/mL*	0.0% (0/3)
127	Rubella virus	1 x 105 TCID50/mL	0.0% (0/3)
128	Salmonella bongori**	NA	NA
129	Salmonella enterica**	NA	NA
130	Serratia marcescens	1 x 106 CFU/mL	0.0% (0/3)
131	Shigella boydii**	NA	NA
132	Shigella flexneri	1 x 106 CFU/mL	0.0% (0/3)
133	Shigella sonnei**	NA	NA
134	Simian Virus type 40	1 x 105 IU/mL	0.0% (0/3)
135	St. Louis encephalitis virus	1 x 105 TCID50/mL	0.0% (0/3)
136	Staphylococcus aureus(MRSA), COL	1 x 106 CFU/mL	0.0% (0/3)
137	Staphylococcus capitis **	NA	NA
138	Staphylococcus epidermidis(MRSE)	1 x 106 CFU/mL	0.0% (0/3)
139	Staphylococcushaemolyticus**	NA	NA
140	Staphylococcus hominis **	NA	NA
141	Staphylococcuslugdunensis **	NA	NA
142	Staphylococcussaprophyticus	1 x 106 CFU/mL	0.0% (0/3)
143	Streptococcus agalactiae	1 x 106 CFU/mL	0.0% (0/3)
144	Streptococcus anginosus **	NA	NA
145	Streptococcus bovis **	NA	NA
146	Streptococcus dysgalactiae	1 x 106 CFU/mL	0.0% (0/3)
147	Streptococcus intermedius	1 x 106 CFU/mL	0.0% (0/3)
148	Streptococcus mutans	1 x 106 CFU/mL	0.0% (0/3)
No.	Organism	Tested Concentration	Agreement with ExpectedResults: % Detection(# Detected/#Tested)
149	Streptococcus oralis**	NA	NA
150	Streptococcus pneumoniae	1 x 106 CFU/mL	0.0% (0/3)
151	Streptococcuspseudopneumoniae**	NA	NA
152	Streptococcus pyogenesZ018	1 x 106 CFU/mL	0.0% (0/3)
153	Streptococcus salivarius	1 x 106 CFU/mL	0.0% (0/3)
154	Streptococcus sanguinis**	NA	NA
155	Toxoplasma gondii	1 x 106 tachyzoites/mL	0.0% (0/3)
156	Treponema pallidum**	NA	NA
157	Tropheryma whipplei**	NA	NA
158	West Nile virus	1 x 105 TCID50/mL	0.0% (0/3)
159	White Blood Cells (HumanGenomic DNA)	1 x 106 cells/mL	0.0% (0/3)

{10}------------------------------------------------

Image /page/10/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a green and blue DNA helix on the left, with the words "DiaSorin" in blue and "Molecular" in green on the right. The logo is simple and modern, and it is likely used to represent the company's focus on molecular diagnostics.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 8 of 25

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Image /page/11/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, with "Molecular" underneath in a lighter green color. The logo is clean and modern, suggesting a focus on biotechnology and molecular diagnostics.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 9 of 25

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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, with "Molecular" underneath in a lighter green color. The logo is clean and modern, suggesting a focus on biotechnology and molecular diagnostics.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 10 of 25

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Image /page/13/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 light green on the bottom line.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 11 of 25

{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, with "Molecular" underneath in green.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 12 of 25

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Image /page/15/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™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 13 of 25

Additional testing was performed in silico due to low concentration of stock and also found not to be cross reactive with VZV.

** Tested in silico due to unavailability of the organism.

INTERFERENCE

The performance of the Simplexa™ VZV Direct assay was evaluated with potentially interfering substances that may be present in CSF samples at the concentrations indicated in the table below. A total of sixteen (16) potentially interfering substances were tested in a low positive VZV sample at approximately 2 times LoD (based on ≥ 95% detection rate) in CSF matrix and assayed in triplicate. No interference was observed at the interferent concentrations indicated. The results are presented in Table 6.

No.	Potential Interferent	VZV Strain	InterferentConcentration	Agreement withExpected Results(#Detected/#Total)
1	Acyclovir	Ellen	9.4 mg/mL	100.0% (3/3)
		9939	9.4 mg/mL	100.0% (3/3)
2	Albumin	Ellen	50 mg/mL	100.0% (3/3)
		9939	50 mg/mL	100.0% (3/3)
3	Bilirubin	Ellen	0.0125 mg/mL	100.0% (3/3)
		9939	0.0125 mg/mL	100.0% (3/3)
4	Casein	Ellen	9.0 mg/mL	100.0% (3/3)
		9939	9.0 mg/mL	100.0% (3/3)
No.	Potential Interferent	VZV Strain	InterferentConcentration	Agreement withExpected Results(#Detected/#Total)
5	Foscarnet	Ellen	0.6 mg/mL	100.0% (3/3)
		9939	0.6 mg/mL	100.0% (3/3)
6	Gamma globulin	Ellen	10.4 mg/mL	100.0% (3/3)
		9939	10.4 mg/mL	100.0% (3/3)
7	Glucose	Ellen	11 mg/mL	100.0% (3/3)
		9939	11 mg/mL	100.0% (3/3)
8	Hemoglobin	Ellen	3.5 mg/mL	100.0% (3/3)
		9939	3.5 mg/mL	100.0% (3/3)
9	Human Genomic DNA	Ellen	72 µg/mL	100.0% (3/3)
		9939	72 µg/mL	100.0% (3/3)
10	Immunoglobulin	Ellen	10 mg/mL	100.0% (3/3)
		9939	10 mg/mL	100.0% (3/3)
11	Lactate	Ellen	2.2 mg/mL	100.0% (3/3)
		9939	2.2 mg/mL	100.0% (3/3)
12	Topical Antiseptic	Ellen	5% (v/v)	100.0% (3/3)
		9939	5% (v/v)	100.0% (3/3)
13	Trans-Isolate Medium	Ellen	50% (v/v)	100.0% (3/3)
		9939	50% (v/v)	100.0% (3/3)
14	UTM	Ellen	50% (v/v)	100.0% (3/3)
		9939	50% (v/v)	100.0% (3/3)
15	White blood cells	Ellen	2x107 WBC/mL	100.0% (3/3)
		9939	2x107 WBC/mL	100.0% (3/3)
16	Whole Blood in EDTA	Ellen	10% (v/v)	100.0% (3/3)
		9939	10% (v/v)	100.0% (3/3)
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
1	NA	Adenovirus A12**	NA	NA
2	NA	Adenovirus B35**	NA	NA
3	Ellen	Adenovirus B7A	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
4	Ellen	Adenovirus C1	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
5	Ellen	Adenovirus C2	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	88.9% (8/9)
6	Ellen	Adenovirus D20	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
7	NA	Adenovirus E4**	NA	NA
8	NA	Adenovirus F41**	NA	NA
9	Ellen	Aspergillus fumigatus	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
10	Ellen	Bacillus cereus	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
11	Ellen	Bacillus subtilis	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
12	Ellen	BK virus	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
13	Ellen	Candida albicans	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
14	Ellen	Candida krusei	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
15	Ellen	Candida parapsilosis	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
16	Ellen	Candida tropicalis	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
17	Ellen	Citrobacter freundii	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
18	Ellen	Citrobacter koseri	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
19	Ellen	Coronavirus 229E	1 x 104 IU/mL*	100.0% (3/3)
	9939		1 x 104 IU/mL*	100.0% (3/3)
20	Ellen	Coronavirus NL63	1 x 104 IU/mL*	100.0% (3/3)
	9939		1 x 104 IU/mL*	100.0% (3/3)
21	Ellen	Coronavirus OC43	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
22	NA	Corynebacteriumstriatum**	NA	NA
23	NA	Corynebacteriumurealyticum**	NA	NA
24	NA	Coxsackievirus A10**	NA	NA
25	Ellen	Coxsackievirus A16	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
26	NA	Coxsackievirus A17**	NA	NA
27	Ellen	Coxsackievirus A21	1 x 104 TCID50/mL*	100.0% (3/3)
	9939		1 x 104 TCID50/mL*	100.0% (3/3)
28	NA	Coxsackievirus A24**	NA	NA
29	NA	Coxsackievirus A6**	NA	NA
30	Ellen	Coxsackievirus A9	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
31	Ellen	Coxsackievirus B1	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
32	Ellen	Coxsackievirus B2	1 x 105 IU/mL	100.0% (3/3)
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
	9939		1 x 105 IU/mL	100.0% (3/3)
33	Ellen	Coxsackievirus B3	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
34	Ellen	Coxsackievirus B4	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
35	Ellen	Coxsackievirus B5	1 x 104 TCID50/mL*	100.0% (3/3)
	9939		1 x 104 TCID50/mL*	100.0% (3/3)
36	Ellen	Cronobacter sakazakii	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
37	NA	Cryptococcus albidus**	NA	NA
38	NA	Cryptococcusamylolentus**	NA	NA
39	NA	Cryptococcus gattii**	NA	NA
40	NA	Cryptococcus laurentii**	NA	NA
41	Ellen	Cryptococcusneoformans	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
42	NA	Cryptococcusuniguttulatus**	NA	NA
43	Ellen	Cytomegalovirus(AD169 Strain)	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
44	Ellen	Dengue virus (Type 1)	1 x 104 UI/mL*	100.0% (3/3)
	9939		1 x 104 IU/mL*	100.0% (3/3)
45	Ellen	Dengue virus (Type 2)	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
46	Ellen	Echovirus 1	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
47	Ellen	Echovirus 11	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
48	NA	Echovirus 18**	NA	NA
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
49	Ellen	Echovirus 4	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
50	Ellen	Echovirus 6	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
51	Ellen	Echovirus 7	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
52	Ellen	Echovirus 9	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
53	Ellen	Encephalomyocarditisvirus	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
54	Ellen	Enterobacter aerogenes	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
55	Ellen	Enterobacter cloacae	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
56	NA	Enterovirus 68**	NA	NA
57	Ellen	Enterovirus 70	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
58	Ellen	Enterovirus 71	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
59	Ellen	Epstein-Barr virus (B95-8 Strain)	1 x 105 copies/mL	100.0% (3/3)
	9939		1 x 105 copies/mL	100.0% (3/3)
60	Ellen	Escherichia coli(O157:H7)	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
61	NA	Escherichia coli K1**	NA	NA
62	NA	Escherichia fergusonii**	NA	NA
63	NA	Escherichia hermanii**	NA	NA
64	NA	Escherichia vulneris **	NA	NA
65	NA	Filobasidium	NA	NA
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
		capsuligenum**
66	Ellen	Haemophilus ducreyi	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
67	NA	Haemophilushaemolyticus**	NA	NA
68	Ellen	Haemophilus influenzaeType A	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
69	Ellen	Haemophilus influenzaeType B	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
70	NA	Haemophilusparahaemolyticus**	NA	NA
71	Ellen	Haemophilusparainfluenzae	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
72	Ellen	Hepatitis A virus	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
73	Ellen	Hepatitis B virus	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
74	Ellen	Hepatitis C virus	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
75	NA	Hepatitis D virus**	NA	NA
76	Ellen	HHV-6A	1 x 105 copies/mL	100.0% (3/3)
	9939		1 x 105 copies/mL	100.0% (3/3)
77	Ellen	HHV-6B	1 x 105 copies/mL	100.0% (3/3)
	9939		1 x 105 copies/mL	100.0% (3/3)
78	Ellen	HHV-7 SB	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	88.9% (8/9)
79	Ellen	HHV-8	1 x 105 copies/mL	100.0% (3/3)
	9939		1 x 105 copies/mL	100.0% (3/3)
80	Ellen	HIV-1 IIIB	1 x 105 IU/mL	100.0% (3/3)
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
	9939		1 x 105 IU/mL	88.9% (8/9)
81	Ellen	HIV-2 NIHZ	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
82	NA	HPeV-1**	NA	NA
83	NA	HPeV-2**	NA	NA
	Ellen		1 x 105 IU/mL	100.0% (3/3)
84	9939	HPeV-3	1 x 105 IU/mL	100.0% (3/3)
85	NA	HPeV-4**	NA	NA
86	NA	HPeV-5**	NA	NA
87	NA	HPeV-6**	NA	NA
	Ellen	HSV-1 (MacIntyre)	1 x 105 TCID50/mL	100.0% (3/3)
88	9939		1 x 105 TCID50/mL	100.0% (3/3)
	Ellen	HSV-2 (G)	1 x 105 IU/mL	100.0% (3/3)
89	9939		1 x 105 IU/mL	100.0% (3/3)
	Ellen	Human Rhinovirus A16	1 x 105 IU/mL	100.0% (3/3)
90	9939		1 x 105 IU/mL	100.0% (3/3)
91	NA	Human Rhinovirus B3**	NA	NA
92	NA	Human Rhinovirus B83**	NA	NA
	Ellen	InfluenzaA/California/7/2009	1 x 105 IU/mL	100.0% (3/3)
93	9939		1 x 105 IU/mL	100.0% (3/3)
	Ellen	Influenza A/H1N1	1 x 105 TCID50/mL	100.0% (3/3)
94	9939		1 x 105 TCID50/mL	100.0% (3/3)
	Ellen	InfluenzaB/Florida/02/2006	1 x 105 IU/mL	100.0% (3/3)
95	9939		1 x 105 IU/mL	100.0% (3/3)
	Ellen	JC virus (MAD-4 strain)	1 x 105 IU/mL	100.0% (3/3)
96	9939		1 x 105 IU/mL	100.0% (3/3)
	Ellen	Klebsiella pneumoniae	1 x 106 CFU/mL	100.0% (3/3)
97	9939		1 x 106 CFU/mL	100.0% (3/3)
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
98	Ellen	La Crosse encephalitisvirus	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
99	NA	Listeria innocua**	NA	NA
100	NA	Listeria ivanovii**	NA	NA
101	Ellen	Listeria monocytogenes	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
102	Ellen	Measles virus	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
103	NA	Morganella morganii**	NA	NA
104	Ellen	Mumps virus	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
105	Ellen	Mycobacteriumtuberculosis genomicDNA	1.6 x 106 copies/mL	100.0% (3/3)
	9939		1.6 x 106 copies/mL	100.0% (3/3)
106	Ellen	Naegleria fowleri	1.78 x 105 cells/mL*	100.0% (3/3)
	9939		1.78 x 105 cells/mL*	100.0% (3/3)
107	Ellen	Neisseria gonorrhoeae	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
108	NA	Neisseria lactamica**	NA	NA
109	Ellen	Neisseria meningitidis(serogroup A)	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
110	NA	Neisseria meningitidis(unencapsulated)**	NA	NA
111	Ellen	Neisseria mucosa	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
112	NA	Neisseria sicca**	NA	NA
113	NA	Pantoea agglomerans**	NA	NA
114	Ellen	Parainfluenza Type 1	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
115	Ellen	Parainfluenza Type 2	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
116	Ellen	Parainfluenza Type 3	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
117	Ellen	Parainfluenza Type 4	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
118	Ellen	Parvovirus B19	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
119	Ellen	Poliovirus (Type 3)	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
120	Ellen	Propionibacterium acnes	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
121	Ellen	Proteus mirabilis Z050	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
122	Ellen	Pseudomonasaeruginosa	1 x 106 CFU/mL	100.0% (3/3)
	9939		1 x 106 CFU/mL	100.0% (3/3)
123	Ellen	Rabies virus	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
124	Ellen	Respiratory syncytialvirus	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
125	Ellen	Rhinovirus 1A	1 x 105 IU/mL	100.0% (3/3)
	9939		1 x 105 IU/mL	100.0% (3/3)
126	Ellen	Rotavirus (Type Wa)	1 x 104 IU/mL*	100.0% (3/3)
	9939		1 x 104 IU/mL*	100.0% (3/3)
127	Ellen	Rubella virus	1 x 105 TCID50/mL	100.0% (3/3)
	9939		1 x 105 TCID50/mL	100.0% (3/3)
128	NA	Salmonella bongori**	NA	NA
129	NA	Salmonella enterica**	NA	NA
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
130	Ellen	Serratia marcescens	1 x 106 CFU/mL	100.0% (3/3)
	9939	Serratia marcescens	1 x 106 CFU/mL	100.0% (3/3)
131	NA	Shigella boydii**	NA	NA
132	Ellen	Shigella flexneri	1 x 106 CFU/mL	100.0% (3/3)
	9939	Shigella flexneri	1 x 106 CFU/mL	100.0% (3/3)
133	NA	Shigella sonnei**	NA	NA
134	Ellen	Simian Virus type 40	1 x 105 IU/mL	100.0% (3/3)
	9939	Simian Virus type 40	1 x 105 IU/mL	100.0% (3/3)
135	Ellen	St. Louis encephalitis virus	1 x 105 TCID50/mL	100.0% (3/3)
	9939	St. Louis encephalitis virus	1 x 105 TCID50/mL	100.0% (3/3)
136	Ellen	Staphylococcus aureus (MRSA), COL	1 x 106 CFU/mL	100.0% (3/3)
	9939	Staphylococcus aureus (MRSA), COL	1 x 106 CFU/mL	100.0% (3/3)
137	NA	Staphylococcus capitis**	NA	NA
138	Ellen	Staphylococcus epidermidis (MRSE)	1 x 106 CFU/mL	100.0% (3/3)
	9939	Staphylococcus epidermidis (MRSE)	1 x 106 CFU/mL	100.0% (3/3)
139	NA	Staphylococcus haemolyticus**	NA	NA
140	NA	Staphylococcus hominis**	NA	NA
141	NA	Staphylococcus lugdunensis**	NA	NA
142	Ellen	Staphylococcus saprophyticus	1 x 106 CFU/mL	100.0% (3/3)
	9939	Staphylococcus saprophyticus	1 x 106 CFU/mL	100.0% (3/3)
143	Ellen	Streptococcus agalactiae	1 x 106 CFU/mL	100.0% (3/3)
	9939	Streptococcus agalactiae	1 x 106 CFU/mL	100.0% (3/3)
144	NA	Streptococcus anginosus**	NA	NA
145	NA	Streptococcus bovis**	NA	NA
146	Ellen	Streptococcus dysgalactiae	1 x 106 CFU/mL	100.0% (3/3)
	9939	Streptococcus dysgalactiae	1 x 106 CFU/mL	100.0% (3/3)
147	Ellen	Streptococcus	1 x 106 CFU/mL	100.0% (3/3)
No.	VZV Strain	Organism	Tested Concentration	Agreement withExpected Results:(# Detected/#Tested)
148	9939	intermedius	1 x 106 CFU/mL	100.0% (3/3)
149	Ellen	Streptococcus mutans	1 x 106 CFU/mL	100.0% (3/3)
	9939	Streptococcus mutans	1 x 106 CFU/mL	100.0% (3/3)
150	NA	Streptococcus oralis **	NA	NA
151	Ellen	Streptococcus pneumoniae	1 x 106 CFU/mL	100.0% (3/3)
	9939	Streptococcus pneumoniae	1 x 106 CFU/mL	100.0% (3/3)
152	NA	Streptococcus pseudopneumoniae **	NA	NA
153	Ellen	Streptococcus pyogenesZ018	1 x 106 CFU/mL	100.0% (3/3)
	9939	Streptococcus pyogenesZ018	1 x 106 CFU/mL	100.0% (3/3)
154	Ellen	Streptococcus salivarius	1 x 106 CFU/mL	100.0% (3/3)
	9939	Streptococcus salivarius	1 x 106 CFU/mL	100.0% (3/3)
155	NA	Streptococcus sanguinis **	NA	NA
156	Ellen	Toxoplasma gondii	1 x 106 Tachyzoites/mL	100.0% (3/3)
	9939	Toxoplasma gondii	1 x 106 Tachyzoites/mL	100.0% (3/3)
157	NA	Treponema pallidum **	NA	NA
158	NA	Tropheryma whipplei **	NA	NA
159	Ellen	West Nile virus	1 x 105 TCID50/mL	100.0% (3/3)
	9939	West Nile virus	1 x 105 TCID50/mL	100.0% (3/3)
160	Ellen	White blood cells(Human genomic DNA)	1 x 106 cells/mL	100.0% (3/3)
	9939	White blood cells(Human genomic DNA)	1 x 106 cells/mL	100.0% (3/3)

Table 6. Simplexa™ VZV Direct Interference

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510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 14 of 25

INHIBITION BY OTHER MICROORGANISMS

The Simplexa™ VZV Direct assay was evaluated by testing the ability to identify VZV virus when other potentially inhibitory organisms are present. The panel of one hundred and sixty (160) potentially inhibitory organisms was individually spiked into a pool with a low concentration at approximately 2 times LoD (based on ≥ 95% detection rate) in CSF. No inhibition by other organisms was observed for VZV at the concentrations indicated in Table 7.

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Image /page/17/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, with "Molecular" underneath in green. The logo is clean and modern, suggesting a company focused on molecular diagnostics or biotechnology.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 15 of 25

Table 7. Simplexa™ VZV Direct Interference

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510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 16 of 25

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Image /page/19/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a DNA helix graphic on the left, with the text "DiaSorin" in blue to the right of the helix. Below "DiaSorin" is the word "Molecular" in green.

510(k) Summary

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Image /page/20/Picture/0 description: The image contains the logo for DiaSorin Molecular. The logo consists of 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 above the word "Molecular" in green.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 18 of 25

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Image /page/21/Picture/0 description: The image shows the logo for DiaSorin Molecular. The word "DiaSorin" is written in dark blue, and the word "Molecular" is written in green. To the left of the words is a green and blue DNA helix.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 19 of 25

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Image /page/22/Picture/0 description: The image contains the logo for DiaSorin Molecular. The word "DiaSorin" is written in a dark blue font, and the word "Molecular" is written in a light green font. To the left of the text is a green and blue DNA helix.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 20 of 25

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Image /page/23/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a DNA helix graphic on the left, with the text "DiaSorin" in blue to the right of the helix. Below "DiaSorin" is the word "Molecular" in green.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 21 of 25

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Image /page/24/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a DNA helix graphic on the left, with the text "DiaSorin" in blue to the right of the helix. Below "DiaSorin" is the word "Molecular" in green.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 22 of 25

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Image /page/25/Picture/0 description: The image contains the logo for DiaSorin Molecular. The logo consists of 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 above the word "Molecular" in green.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 23 of 25

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Image /page/26/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 a dark blue, sans-serif font, with the word "Molecular" underneath in a lighter green color.

510(k) Summary

Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 24 of 25

Any organisms with low titer stock were analyzed in silico by BLAST (Basic Local Alignment Search Tool, NCBI, NIH). ** Tested in silico due to unavailability of the organism.

CARRY-OVER CONTAMINATION

An amplification carry-over for the Simplexa™ assays has been assessed. The study was designed by alternately placing high positive and negative samples on each disc. No evidence of carry-over contamination was observed.

EXPECTED VALUES

The prevalence of VZV as determined by the Simplexa™ VZV Direct assay in a multi-site clinical study with prospectively collected specimens is shown in Table 8 below. The expected values for VZV per site varied between 0% and 7.4%. The overall prevalence for all sites was 2.2% (fourteen of six hundred thirty-seven (14/637).

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Image /page/27/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, with the word "Molecular" underneath in light green. The logo is clean and modern, suggesting a company focused on molecular diagnostics or related fields.

510(k) Summary Simplexa™ VZV Direct Catalog No. MOL3650 Simplexa™ VZV Positive Control Pack Catalog No. MOL3660 May 3, 2019 Page 25 of 25

Table 8. Prospective VZV prevalence as determined by Simplexa™ VZV Direct

Site ID	Total	Simplexa™ VZV DirectPositive Samples	VZV Prevalence
1	199	3	1.5%
3	181	2	1.1%
6	27	2	7.4%
7	71	0	0%
8	20	0	0%
10	139	7	5%
All	637	14	2.2%

Regulation Number and Section

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

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