(88 days)
The DiaSorin Molecular Simplexa™ VZV Swab Direct assay is intended for use on the LIAJSON® MDX instrument for the qualitative detection of varicella-zoster virus (VZV) DNA present in cutaneous and mucocutaneous lesion swabs from patients with signs and symptoms of VZV infection. This test is intended as an aid in the diagnosis of VZV infection. Negative results do not preclude VZV infection and should not be used as the sole basis for treatment or other patient management decisions.
The Simplexa™ VZV Positive Control Pack is intended to be used as a control with the Simplexa™ VZV Direct kit and the Simplexa™ VZV Swab Direct kit on the LIAISON® MDX Instrument. It is not intended for use with other assays or systems.
The Simplexa™ VZV Swab Direct assay is a real-time PCR system that enables the direct amplification and detection of VZV DNA from unprocessed cutaneous and mucocutaneous lesion swab specimens without nucleic acid extraction. The system consists of the Simplexa™ VZV Swab Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc (DAD) and associated accessories.
In the Simplexa™ VZV Swab 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.
This document describes the validation of the DiaSorin Molecular Simplexa™ VZV Swab Direct assay. The assay is intended for the qualitative detection of varicella-zoster virus (VZV) DNA directly from cutaneous and mucocutaneous lesion swabs.
1. Table of Acceptance Criteria and Reported Device Performance:
The document implicitly defines acceptance criteria through the results of the clinical and analytical studies, which consistently demonstrate high agreement and sensitivity/specificity.
| Study Type | Acceptance Criterion (Implicit) | Reported Device Performance |
|---|---|---|
| Clinical Agreement (Prospective) | High Sensitivity and Specificity with Composite Reference Method (CRM) | All (N=452): Sensitivity 97.8% (95% CI: 92.2% - 99.4%), Specificity 99.2% (95% CI: 97.6% - 99.7%) Mucocutaneous (N=179): Sensitivity 87.5% (95% CI: 52.9% - 97.8%), Specificity 100.0% (95% CI: 97.8% - 100.0%) Cutaneous (N=245): Sensitivity 98.8% (95% CI: 93.3% - 99.8%), Specificity 98.2% (95% CI: 94.8% - 99.4%) |
| Clinical Agreement (Retrospective) | High Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) with Composite Reference Method 2 (CRM 2) | All (N=180): PPA 98.4% (95% CI: 91.4% - 99.7%), NPA 99.2% (95% CI: 95.4% - 99.9%) Mucocutaneous (N=73): PPA 90.0% (95% CI: 59.6% - 98.2%), NPA 100.0% (95% CI: 94.3% - 100.0%) Cutaneous (N=107): PPA 100.0% (95% CI: 93.1% - 100.0%), NPA 98.2% (95% CI: 90.4% - 99.7%) |
| Clinical Agreement (Contrived) | 100% PPA and NPA with CRM 2 | All (N=120): PPA 100.0% (95% CI: 94.0% - 100.0%), NPA 100.0% (95% CI: 94.0% - 100.0%) |
| Reproducibility | 100% Agreement with Expected Results across sites, runs, and operators for positive and negative controls. Consistent Ct values across sites. | All positive samples (LP, MP for both strains, and PC): 100.0% agreement with expected results (90/90 replicates). Negative Control (UTM): 0.0% agreement with expected results (0/90 replicates). Low %CV for Ct values indicating good precision (e.g., typically <5%). |
| Analytical Sensitivity (LoD) | Lowest concentration detected ≥95% of the time. | VZV Strain 9939: 0.77 TCID50/mL (800 Copies/mL) VZV Strain Ellen: 0.054 TCID50/mL (3500 Copies/mL) |
| Analytical VZV Strain Reactivity | Detection of other VZV strains at specified concentrations. | All tested VZV strains (Strain 82, 275, 1700, Isolate A, B, D) were detected 3/3 times at concentrations referenced (e.g., 0.82-2.47 TCID50/mL). In silico BLAST analysis predicted detection of 178 additional VZV strains. |
| Cross-Reactivity | No cross-reactivity with closely related organisms, organisms causing similar symptoms, or common commensals. | No cross-reactivity observed with 99 different bacteria, viruses, parasites, and fungi tested at specified concentrations. In silico analysis confirmed no cross-reactivity for three additional organisms. |
| Inhibition by Other Microorganisms | No inhibition of VZV detection in the presence of other microorganisms. | No inhibition observed for the detection of VZV Ellen or 9939 strains when individually spiked with 99 potential inhibitory organisms at specified concentrations. |
| Interference | No interference with VZV detection in the presence of common endogenous and exogenous substances. | No interference observed for the detection of VZV Ellen or 9939 strains when individually spiked with 45 potentially interfering substances at specified concentrations. |
| Carry-over Contamination | No evidence of carry-over contamination. | No evidence of carry-over contamination observed in a study using the Simplexa™ Flu A/B & RSV Direct assay (deemed applicable). |
2. Sample sizes used for the test set and the data provenance:
- Prospective Study: 452 cutaneous and mucocutaneous specimens.
- Data Provenance: Collected from 10 collection sites across the USA (November 2018 - May 2019). This is prospective data.
- Retrospective Study: 60 positive and 120 negative (masked) cutaneous and mucocutaneous swab specimens. Total of 180 samples.
- Data Provenance: Retrospective and blinded, specific country not mentioned but implied to be related to DiaSorin Molecular (Cypress, California, USA).
- Contrived Sample Study: 60 contrived positive specimens (30 VZV Ellen, 30 VZV 9939) and 60 masked negative specimens. Total of 120 samples.
- Data Provenance: Contrived samples are laboratory-prepared.
3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
The document does not specify the "number of experts" or their specific "qualifications" (e.g., "radiologist with 10 years of experience") for establishing the ground truth. Instead, it describes a "Composite Reference Method (CRM)" for the prospective study and "Composite Reference Method 2 (CRM 2)" for the retrospective and contrived studies.
- Prospective Study (CRM): The ground truth involved a three-part composite reference method:
- VZV direct stain fluorescent antibody (DSFA) and/or culture isolation with direct fluorescent antibody (DFA) performed by one testing site.
- Two validated VZV polymerase chain reaction (PCR) assays followed by bi-directional sequencing performed by another testing site.
- Retrospective and Contrived Studies (CRM 2): The ground truth utilized a two out of three outcome from:
- One FDA Cleared NAAT PCR assay for VZV (performed by one external site).
- Two validated VZV PCR assays followed by bi-directional sequencing (performed by different DiaSorin Molecular operators).
While these methods represent established laboratory diagnostic techniques, the specific number and qualifications of individuals performing or interpreting these reference methods are not detailed.
4. Adjudication method (e.g. 2+1, 3+1, none) for the test set:
- Prospective Study: The document doesn't explicitly state an adjudication method like "2+1" or "3+1." It mentions a "three (3) part composite reference method." The results presented ("sensitivity" and "specificity") suggest a consensus or majority rule type of adjudication where the CRM's outcome defines the true positive/negative.
- Retrospective and Contrived Studies: The ground truth (CRM 2) explicitly used a "two (2) out of three (3) outcome" from the reference methods, which is a form of majority rule adjudication.
5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
This is a diagnostic device (molecular assay) for the detection of VZV DNA, not an AI-powered image analysis tool for human readers. Therefore, a multi-reader multi-case (MRMC) comparative effectiveness study with human readers (or improvement with AI assistance) is not applicable and was not performed. The device itself is a standalone diagnostic test performed by laboratory personnel.
6. If a standalone (i.e. algorithm only without human-in-the loop performance) was done:
Yes, the studies described are standalone performance studies of the Simplexa™ VZV Swab Direct assay. The assay is an in vitro diagnostic test that provides a qualitative result (positive or negative for VZV DNA) based on real-time PCR, effectively operating as an "algorithm only" in the sense that its output is directly read without requiring further human interpretive modifications beyond standard laboratory operating procedures.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
The ground truth used was a composite reference method consisting of:
- For the Prospective Study:
- VZV direct stain fluorescent antibody (DSFA) and/or culture isolation with direct fluorescent antibody (DFA).
- Two validated VZV polymerase chain reaction (PCR) assays followed by bi-directional sequencing.
- For the Retrospective and Contrived Studies:
- One FDA Cleared NAAT PCR assay for VZV.
- Two validated VZV PCR assays followed by bi-directional sequencing.
This combination indicates a sophisticated, multi-modal ground truth incorporating both direct viral detection/culture and highly sensitive molecular methods with genetic confirmation (sequencing). This is a robust form of ground truth for pathogen detection.
8. The sample size for the training set:
The document does not describe a "training set" in the context of machine learning. The studies described are validation studies for a molecular diagnostic assay. The assay's design (primers, probes, reaction conditions) would have been developed/optimized prior to these validation studies, but this process is not typically referred to as "training" in the same way as for AI/ML models.
9. How the ground truth for the training set was established:
As there is no "training set" in the AI/ML sense, this question is not applicable. The ground truth for the validation (test) sets was established using the Composite Reference Methods described in point 7.
{0}------------------------------------------------
November 26, 2019
Image /page/0/Picture/1 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health and 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 Sharon Young Principal Regulatory Affairs Specialist 11331 Valley View Street Cypress, California 90630
Re: K192376
Trade/Device Name: Simplexa VZV Swab Direct. Simplexa VZV Positive Control Pack Regulation Number: 21 CFR 866.3309 Regulation Name: Herpes Virus Nucleic Acid-Based Cutaneous and Mucocutaneous Lesion Panel Regulatory Class: Class II Product Codes: PGI, PMN Dated: August 28, 2019 Received: August 30, 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 requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part
{1}------------------------------------------------
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 medical devices and radiation-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,
Maria Ines Garcia, Ph.D. Branch Chief Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
{2}------------------------------------------------
Image /page/2/Picture/1 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, 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) Sui Simplexa™ VZV Swa Simplexa™ VZV Positive Control Pack Page 1 of 15
| Applicant | DiaSorin Molecular LLC.11331 Valley View StreetCypress, California 90630USA |
|---|---|
| Establishment Registration No. | 2023365 |
| Contact Person | Sharon YoungPrincipal Regulatory Affairs Specialisttel 562.240.6680fax 562.240.6530Sharon.Young@DiaSorin.com |
| Summary Date | November 18, 2019 |
| Proprietary Name | Simplexa™ VZV Swab Direct and Simplexa™ VZV Positive ControlPack |
| US Product Codes/Names andRegulation Numbers | PGI / Herpes virus nucleic acid-based cutaneous andmucocutaneous lesion panel 21 CFR § 866.3309PMN / Assayed external control material for microbiology nucleicacid amplification (NAT) assays 21 CFR § 866.3920 |
| Classification | Class II |
| Predicate Device | Solana® HSV 1+2/VZV Assay (K162451) |
Intended Use
Simplexa™ VZV Direct
The DiaSorin Molecular Simplexa™ VZV Swab Direct assay is intended for use on the LIAJSON® MDX instrument for the qualitative detection of varicella-zoster virus (VZV) DNA present in cutaneous and mucocutaneous lesion swabs from patients with signs and symptoms of VZV infection. This test is intended as an aid in the diagnosis of VZV infection. Negative results do not preclude VZV infection and should not be used as the sole basis for treatment or other patient management decisions.
Simplexa™ VZV Positive Control Pack
The Simplexa™ VZV Positive Control Pack is intended to be used as a control with the Simplexa™ VZV Direct kit and the Simplexa™ VZV Swab Direct kit on the LIAISON® MDX Instrument. It is not intended for use with other assays or systems.
Device Description
The Simplexa™ VZV Swab Direct assay is a real-time PCR system that enables the direct amplification and detection of VZV DNA from unprocessed cutaneous and mucocutaneous lesion swab specimens without nucleic acid extraction. The system consists of the Simplexa™ VZV Swab Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc (DAD) and associated accessories.
In the Simplexa™ VZV Swab 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.
{3}------------------------------------------------
Image /page/3/Picture/0 description: The image shows 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, with "Molecular" underneath in green. The logo is clean and modern, suggesting a company focused on molecular diagnostics or related fields.
510(k) Summary Simplexa™ VZV Swab Direct Simplexa™ VZV Positive Control Pack Page 2 of 15
Simplexa™ VZV Direct Kit
| Component Name | REF | EC SYMBOL ON LABEL | Abbreviated Name | Cap Color | Number of Vials | Reactions per Vial/Kit | Volume per Vial |
|---|---|---|---|---|---|---|---|
| Simplexa™ VZV Swab Direct Reaction Mix | MOL3656 | REAG C | RM | Black | 24 | 1/24 | 50 μL |
Simplexa™ VZV Direct Components and Descriptions
| Kit Component | Contents | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Simplexa™ VZVSwab DirectReaction Mix (RM) | DNA polymerase, buffer, dNTPs, template DNA (Internal Control), dye-labeled fluorescentprobes and primers specific for detection of VZV Swab Direct and for the DNA InternalControl. | |||||||||||||||
| Target ProbeFluorophore(Dye) Excitation (nm) Emission (nm) Targeted Gene VZV FAM 495 520 VZV DNApolymerase InternalControlDNA (IC) Q670 644 670 N/A | ||||||||||||||||
| Simplexa™ VZVSwab Direct KitBarcode Card | Assay specific parameters and lot information. |
Simplexa™ VZV Positive Control Pack Component and Description
| Component Name | REF | Description | Cap Color | Numberof Vials | Reactions perVial/Kit | Volumeper Vial |
|---|---|---|---|---|---|---|
| Simplexa™ VZV DirectPositive Control | MOL3661 | Inactivatedvaricella-zostervirus | Red | 10 | 1/10 | 50 μL |
Materials Supplied Separately
Direct Amplification Disc Kit Direct Amplification Discs for use on the LIAISON® MDX
{4}------------------------------------------------
Image /page/4/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 Swab Direct
Simplexa™ VZV Positive Control Pack Page 3 of 15
Comparison to Predicate Device
| Comparison toPredicate Device | Predicate Device:Solana® HSV 1+2/VZV Assay (K162451) | Candidate Device:Simplexa™ VZV Direct and Simplexa™ VZVPositive Control Pack |
|---|---|---|
| Product Code | PGI | Same |
| RegulationNumber | 21 CFR 866.3309 – Herpes virus nucleicacid-based cutaneous and mucocutaneouslesion panel | Same |
| OrganismDetected | Varicella-zoster virus | Same |
| Measurand | DNA from varicella-zoster virus | Same |
| Intended Use | The Solana® HSV 1+2/VZV Assay is an in vitro diagnostic test, using isothermal amplification technology (helicase-dependent amplification, HDA), for the qualitative detection and differentiation of herpes simplex virus type 1, herpes simplex virus type 2, and varicella-zoster virus DNA isolated and purified from cutaneous or mucocutaneous lesion samples obtained from symptomatic patients suspected of active herpes simplex virus 1, herpes simplex virus 2 and/or varicella-zoster infection.The Solana® HSV 1+2/VZV Assay is intended to aid in the diagnosis of herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus active cutaneous or mucocutaneous infections.Negative results do not preclude herpes simplex virus 1, herpes simplex virus 2 and varicella-zoster virus infections and should not be used as the sole basis for diagnosis, treatment or other management decisions.The Solana® HSV 1+2/VZV Assay is intended for use only with the Solana® instrument. | The DiaSorin Molecular Simplexa™ VZV Swab Direct assay is intended for use on the LIAISON® MDX instrument for the qualitative detection of varicella-zoster virus (VZV) DNA present in cutaneous and mucocutaneous lesion swabs from patients with signs and symptoms of VZV infection. This test is intended as an aid in the diagnosis of VZV infection. Negative results do not preclude VZV infection and should not be used as the sole basis for treatment or other patient management decisions. |
| AutomatedSystem (Sampleto Answer) | Yes | Yes |
| Instrumentation | Solana® Instrument | LIAISON® MDX |
{5}------------------------------------------------
Image /page/5/Picture/0 description: The image contains the logo for DiaSorin Molecular. The logo consists of a stylized DNA double helix on the left, colored in shades of green and blue. To the right of the helix is the text "DiaSorin" in a bold, dark blue font, with "Molecular" underneath in a lighter green color. The overall design is clean and professional, suggesting a company focused on molecular diagnostics or related fields.
CLINICAL AGREEMENT
The performance of the Simplexa™ VZV Swab Direct assay was established in a clinical study that included three (3) cohorts based on sample status. Specifically, prospective and retrospective cutaneous and mucocutaneous swab samples from human patients with signs and symptoms of VZV infection, as well as contrived samples, were tested in the clinical agreement study.
Prospective Study
A total of four hundred fifty-two (452) cutaneous and mucocutaneous prospective specimens were collected from ten (10) collection sites across the USA during the clinical study (November 2018 - May 2019). The specimens were taken from anorectal, genital, nasal, ocular, oral, skin and urethral locations of the body. The age of the patients ranged from one (1) month to greater than 60 (>60) years of age. Of these specimens, sixty-two point four percent (62.4%) of the specimens were from female patients and thirty-seven point six percent (37,6%) of the specimens were from male patients. Ten (10) testing sites performed the Simplexa™ VZV Swab Direct assay on enrolled specimens and shipped the specimens to two (2) comparator testing sites to test against a three (3) part composite reference method (CRM). The three part CRM consisted of VZV direct stain fluorescent antibody (DSFA) and/or culture isolation with direct fluorescent antibody (DFA) and two (2) validated VZV polymerase chain reaction (PCR) assays followed by bi-directional sequencing. The comparator testing was performed by two (2) sites. One (1) testing site performed the VZV DSFA and/or culture isolation with DFA and another testing site conducted the two (2) validated VZV PCR assays testing followed by bi-directional sequencing. The results of the study are presented in Table 1a.
| Composite ReferenceMethod (CRM) | ||||||||
|---|---|---|---|---|---|---|---|---|
| ProspectiveStudy | +Simplexa™VZV SwabDirect | -Simplexa™VZV SwabDirect | Total | Sensitivity95% CI | Specificity95% CI | |||
| + | - | + | - | |||||
| Mucocutaneous | 7 | 1a | 0 | 171 | 179 | 87.5% (7/8)52.9% - 97.8% | 100.0% (171/171)97.8% - 100.0% | |
| Cutaneous | 79 | 1b | 3 | 162 | 245 | 98.8% (79/80)93.3% - 99.8% | 98.2% (162/165)94.8% - 99.4% | |
| Unknown | 1 | 0 | 0 | 27 | 28 | 100.0% (1/1)20.7% - 100.0% | 100.0% (27/27)87.5% - 100.0% | |
| All | 87 | 2 | 3 | 360 | 452 | 97.8% (87/89)92.2% -99.4 % | 99.2% (360/363)97.6% - 99.7% |
| Table 1a. Simplexa™ VZV Swab Direct Prospective Aqreement Results |
|---|
The discordant negative mucocutaneous result is from an oral lesion sample was negative by the Simplexa™ VZV Swab Direct, DSFA/DFA and by the sites routine culture testing. The sample was positive by the two (2) PCR/Bi-directional sequencing assays.
The discordant negative cutaneous result is from a skin lesion sample. The sample was negative by the Simplexa™ VZV Swab Direct, DSFA/DFA testing. The sample was positive by the 2 PCR/Bi-dir sequencing assays
Cl = Confidence Interval. The 95% confidence intervals (Cl) were calculated following Wilson Score method.
Retrospective Study
A total of sixty (60) cutaneous and mucocutaneous retrospective swab specimens in UTM were blinded and randomized with one hundred twenty (120) negative masked specimens prior to being tested by Simplexa™ VZV Swab Direct assay. The Composite Reference Method 2 (CRM 2) utilized a two (2) out of three (3) outcome from one (1) FDA Cleared NAAT PCR assay for VZV and two (2) validated VZV
{6}------------------------------------------------
Image /page/6/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.
PCR assays followed by bi-directional sequencing. The FDA Cleared NAAT was performed by one (1) external site. DiaSorin Molecular performed the Simplexa™ VZV Swab Direct testing and different DiaSorin Molecular operators performed the two (2) validated VZV PCR assays followed by bi-directional sequencing. The positive and negative percent agreement (PPA and NPA) results of the study are presented in Table 1b.
| RetrospectiveStudy | Composite ReferenceMethod 2(CRM 2) | Total | PPA95% CI | NPA95% CI | |||
|---|---|---|---|---|---|---|---|
| +Simplexa™VZV SwabDirect | -Simplexa™VZV SwabDirect | ||||||
| + | - | + | - | ||||
| Mucocutaneous | 9 | 1c | 0 | 63 | 73 | 90.0% (9/10)59.6% - 98.2% | 100.0% (63/63)94.3% - 100.0% |
| Cutaneous | 52 | 0 | 1 | 54 | 107 | 100.0% (52/52)93.1% - 100.0% | 98.2% (54/55)90.4% - 99.7% |
| All | 61 | 1 | 1 | 117 | 180 | 98.4% (61/62)91.4% - 99.7% | 99.2% (117/118)95.4% - 99.9% |
Table 1b. Simplexa™ VZV Swab Direct Retrospective Agreement Results
The discordant neqative mucocutaneous result is from an oral lesion sample. The sample was negative by the Simplexa™ VZV Swab Direct and NAAT testing. The sample was positive by the two (2) PCR/Bi directional sequencing assays.
Cl = Confidence Interval. The 95% confidence intervals (CI) were calculated following Wilson Score method.
Contrived Sample Study
A total of sixty (60) contrived positive specimens in individual negative UTM mucocutaneous swab matrix were blinded and randomized with sixty (60) masked negative UTM mucocutaneous specimens prior to being tested by Simplexa™ VZV Swab Direct assay. The results were compared with a two (2) out of three (3) outcome from one (1) FDA Cleared NAAT assay and two (2) validated VZV PCR assays followed by bidirectional sequencing (Composite Reference Method 2 or CRM 2). Of the sixty (60) contrived specimens, thirty (30) were spiked with VZV Ellen strain and thirty (30) were spiked with VZV 9939 strain at different known concentrations across the detection range. The results are presented in Table 1c.
| Table 1c. Simplexa™ VZV Swab Direct Contrived Agreement Results | |||
|---|---|---|---|
| Composite ReferenceMethod 2(CRM 2) | |||||||
|---|---|---|---|---|---|---|---|
| ContrivedMucocutaneousSamples | +Simplexa™VZV SwabDirect | -Simplexa™VZV SwabDirect | Total | PPA95% CI | NPA95% CI | ||
| + | - | + | - | ||||
| Ellen | 30 | 0 | 0 | 0 | 30 | 100.0% (30/30)88.6% - 100.0% | N/A |
| 9939 | 30 | 0 | 0 | 0 | 30 | 100.0% (30/30)88.6% - 100.0% | N/A |
| Negative | 0 | 0 | 0 | 60 | 60 | N/A | 60/60 (100%)94.0% -100.0% |
| All | 60 | 0 | 0 | 60 | 120 | 100.0% (60/60)94.0% - 100.0% | 100.0% (60/60)94.0% - 100.0% |
Cl = Confidence Interval. The 95% confidence intervals (CI) were calculated following Wilson Score method.
{7}------------------------------------------------
Image /page/7/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA helix in green and blue on the left. To the right of the helix are the words "DiaSorin" in dark blue on the top line and "Molecular" in green on the bottom line.
510(k) Sun marv Simplexa™ VZV Swab Direct Simplexa™ VZV Positive Control Pack Page 6 of 15
REPRODUCIBILITY
Reproducibility for the Simplexa™ VZV Swab Direct assay was evaluated at three (3) investigative sites to assess the device's inter-site, inter/intra-day and inter/intra-assay reproducibility. Each of the laboratories tested a sample panel consisting of Simplexa™ VZV Swab Direct Positive Control. No Template Control, 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) at 2X LoD and a medium positive (MP) at 4X LoD for each VZV strain. Each sample panel member was tested in triplicate per run, for two (2) runs per day by two (2) different operators at each site. Therefore, a total of ninety (90) replicates [three (3) replicates X two (2) runs X five (5) days X three (3) sites] were tested for each sample panel member. A total of six (6) LIAISON® MDX instruments [two (2) per site] were used to evaluate the reproducibility study. The combined results for all sites are presented in Table 2.
| Summary of VZV Qualitative Results and VZV Ct Values ± SD (%CV) | ||||||||
|---|---|---|---|---|---|---|---|---|
| Site 1 | Site 2 | Site 4 | All Sites | |||||
| Sample | % Agreement With Expected Results | Detected Mean Ct ± SD (%CV) | % Agreement With Expected Results | Detected Mean Ct ± SD (%CV) | % Agreement With Expected Results | Detected Mean Ct ± SD (%CV) | % Agreement With Expected Results | Detected Mean Ct ± SD (%CV) |
| 9939 LP | 100.0%(30/30) | 36.6 ± 1.12(3.1%) | 100.0%(30/30) | 36.8 ± 0.68(1.9%) | 100.0%(30/30) | 36.4 ± 0.83(2.3%) | 100.0%(90/90) | 36.6 ± 0.9(2.5%) |
| 9939 MP | 100.0%(30/30) | 35.8 ± 0.86(2.4%) | 100.0%(30/30) | 35.7 ± 0.54(1.5%) | 100.0%(30/30) | 35.3 ± 0.78(2.2%) | 100.0%(90/90) | 35.6 ± 0.76(2.1%) |
| Ellen LP | 100.0%(30/30) | 35.4 ± 1.22(3.4%) | 100.0%(30/30) | 34.5 ± 1.77(5.1%) | 100.0%(30/30) | 35.0 ± 0.56(1.6%) | 100.0%(90/90) | 35.0 ± 1.32(3.8%) |
| Ellen MP | 100.0%(30/30) | 34.5 ± 0.65(1.9%) | 100.0%(30/30) | 34.5 ± 0.47(1.4%) | 100.0%(30/30) | 33.5 ± 1.3(3.9%) | 100.0%(90/90) | 34.1 ± 0.99(2.9%) |
| UTM(NTC) | 0.0%(0/30) | 0.0 ± 0.00(N/A) | 0.0%(0/30) | 0.0 ± 0.00(N/A) | 0.0%(0/30) | 0.0 ± 0.00(N/A) | 0.0%(0/90) | 0.0 ± 0.00(N/A) |
| PC | 100.0%(30/30) | 30.2 ± 0.74(2.5%) | 100.0%(30/30) | 30.4 ± 0.59(1.9%) | 100.0%(30/30) | 29.7 ± 0.86(2.9%) | 100.0%(90/90) | 30.1 ± 0.79(2.6%) |
Table 2. Simplexa™ VZV Swab Direct Reproducibility
ANALYTICAL SENSITIVITY/LIMIT OF DETECTION
The limit of detection (LoD) was determined for the Simplexa™ VZV Swab Direct assay using quantified stocks of two (2) VZV strains (Ellen and 9939) in a pool of cutaneous and mucocutaneous lesion swab sample types in UTM using forty-eight (48) replicates. The LoD was determined to be the lowest concentration that could be detected positive ≥95% of the time. The LoD results are presented in Table 3.
{8}------------------------------------------------
Image /page/8/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the DNA graphic are the words "DiaSorin" in dark blue on the top line and "Molecular" in green on the bottom line. The logo is clean and modern, suggesting a company focused on molecular diagnostics or related fields.
Table 3. Simplexa™ VZV Swab Direct Limit of Detection
| VZV Strain | LoD (TCID50/mL) | LoD(Copies /mL) |
|---|---|---|
| 9939 | 0.77 | 800 |
| Ellen | 0.054 | 3500 |
ANALYTICAL VZV STRAIN REACTIVITY
Analytical VZV strain reactivity was evaluated with cutaneous and mucocutaneous lesion swab specimens with reference to LoD for the Simplexa™ VZV Swab Direct assay. Quantified viral material was spiked into negative matrix using a single dilution and assayed in triplicate. The Simplexa™ VZV Swab Direct assay was able to detect other strains of VZV. The results are presented in Table 4. In addition to the strains that were physically tested, in silico BLAST analysis demonstrated that the assay is expected to detect at least one hundred seventy-eight (178) additional VZV strains.
| VZV Strain | Concentration (TCID50/mL) | Qualitative Result(# Detected/# Tested) |
|---|---|---|
| VZV Strain 82 | 0.82 | 3/3 |
| VZV Strain 275 | 0.82 | 3/3 |
| VZV Strain 1700 | 2.47 | 3/3 |
| VZV Isolate A | 0.82 | 3/3 |
| VZV Isolate B | 0.82 | 3/3 |
| VZV Isolate D | 0.82 | 3/3 |
Table 4. Simplexa™ VZV Swab Direct Analytical Reactivity With VZV Strains
CROSS-REACTIVITY (Analytical Specificity)
The Simplexa™ VZV Swab Direct assay's analytical specificity was evaluated by testing the ability of the assay to exclusively identify VZV with no cross-reactivity to organisms that are closely related, or cause similar clinical symptoms or that could be found in cutaneous and mucocutaneous lesion swab specimens. Analytical specificity/cross-reactivity was tested with ninety-nine (99) different bacteria, viruses, parasites and fungi and assayed in triplicate. No cross-reactivity was observed with the ninety-nine (99) organisms. The organisms and the concentration at which these were tested are presented in Table 5.
{9}------------------------------------------------
Image /page/9/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in a dark blue, sans-serif font, with the word "Molecular" underneath in a lighter green color.
510(k) Summary Simplexa™ VŽV Swab Direct
Simplexa™ VZV Positive Control Pack Page 8 of 15
Table 5. Simplexa™ VZV Swab Direct Cross-Reactivity
| Organism | Tested Concentration | Organism | Tested Concentration |
|---|---|---|---|
| Acholeplasma laidlawi(genomic DNA) | 1 x 106 copies/mL | Human genomic DNA | 1 x 106 copies/mL |
| Acinetobactercalcoaceticus | 1 x 106 CFU/mL | Human metapneumovirusA1 | 1 x 105 TCID50/mL |
| Acinetobacter Iwoffi | 1 x 106 CFU/mL | Human Papilloma Virus18 | 1 x 105 copies/mL |
| Adenovirus 7A | 1 x 105 TCID50/mL | InfluenzaA/California/7/2009 | 1 x 105 TCID50/mL |
| Bacteroides fragilis | 1 x 106 CFU/mL | InfluenzaB/Florida/02/2006 | 1 x 105 TCID50/mL |
| Bordetella bronchiseptica | 1 x 106 CFU/mL | Klebsiella pneumoniae | 1 x 106 CFU/mL |
| Bordetella pertussis | 1 x 106 CFU/mL | Lactobacillus acidophilus | 1 x 106 CFU/mL |
| Borrelia burgdorferi(genomic DNA) | 1 x 106 copies/mL | Legionella pneumophila | 1 x 106 CFU/mL |
| Candida albicans | 1 x 106 CFU/mL | Measles virus | 1 x 105 TCID50/mL |
| Candida glabrata | 1 x 106 CFU/mL | Mobiluncus curtisii | 1 x 106 CFU/mL |
| Candida guilliermondii | 1 x 106 CFU/mL | Mobiluncus mulieris | 1 x 106 CFU/mL |
| Candida krusei | 1 x 106 CFU/mL | Moraxella cartarrhalis | 1 x 106 CFU/mL |
| Candida lusitaniae | 1 x 106 CFU/mL | Mumps virus | 1 x 105 TCID50/mL |
| Candida parapsilosis | 1 x 106 CFU/mL | Mycoplasma genitalium | 1 x 106 CCU/mL |
| Candida tropicalis | 1 x 106 CFU/mL | Mycoplasma hominis | 1 x 106 CCU/mL |
| Chlamydia trachomatis | 1 x 106 IFU/mL | Mycoplasma hyorhinis | 1 x 106 CCU/mL |
| Chlamydophilapneumoniae | 1 x 106 IFU/mL | Mycoplasma orale | 1 x 106 CCU/mL |
| Clostridium difficile | 1 x 106 CFU/mL | Mycoplasma pneumoniae | 1 x 106 CCU/mL |
| Clostridium perfringens | 1 x 106 CFU/mL | Mycoplasma salivarium | 1 x 106 CCU/mL |
| Clostridium sordellii | 1 x 106 CFU/mL | Neisseria gonorrhoeae | 1 x 106 CFU/mL |
| Coronavirus OC43 | 1 x 105 TCID50/mL | Neisseria meningitidis | 1 x 106 CFU/mL |
| Corynebacteriumdiphtheriae | 1 x 106 CFU/mL | Parainfluenza Type 1 | 1 x 105 TCID50/mL |
| Corynebacteriumgenitalium | 1 x 106 CFU/mL | Parainfluenza Type 2 | 1 x 105 TCID50/mL |
| Coxsackievirus B1 | 1 x 105 TCID50/mL | Parainfluenza Type 3 | 1 x 105 TCID50/mL |
| Coxsackievirus B4 | 1 x 105 TCID50/mL | Parainfluenza Type 4 | 1 x 105 TCID50/mL |
| Cytomegalovirus (AD169strain) | 1 x 105 TCID50/mL | Prevotellamelaninogenica | 1 x 106 CFU/mL |
| Cytomegalovirus (Townestrain) | 1 x 105 TCID50/mL | Proteus mirabilis | 1 x 106 CFU/mL |
| Echovirus 11 | 1 x 105 TCID50/mL | Proteus vulgaris | 1 x 106 CFU/mL |
| Enterobacter cloacae | 1 x 106 CFU/mL | Pseudomonas aeruginosa | 1 x 106 CFU/mL |
| Enterococcus faecalisvanB | 1 x 106 CFU/mL | RSV A Long | 1 x 105 TCID50/mL |
| Organism | Tested Concentration | Organism | Tested Concentration |
| Enterococcus faecium | 1 x 106 CFU/mL | RSV B Washington | 1 x 105 TCID50/mL |
| Enterovirus 70 | 1 x 105 TCID50/mL | Rubella Virus | 1 x 105 TCID50/mL |
| Enterovirus 71 | 1 x 105 TCID50/mL | Salmonella enteritidis(genomic DNA) | 1 x 106 copies/mL |
| Epstein Barr Virus (B95-8strain) | 1 x 105 copies/mL | Salmonella typhimurium | 1 x 106 CFU/mL |
| Escherichia coli O15:H7 | 1 x 106 CFU/mL | Serratia marcescens | 1 x 106 CFU/mL |
| Fusobacterium nucleatum | 1 x 106 CFU/mL | Simian Virus type 40 | 1 x 105 TCID50/mL |
| Gardnerella vaginalis | 1 x 106 CFU/mL | Staphylococcus aureus(MRSA), ATCC 700699 | 1 x 106 CFU/mL |
| Haemophilus ducreyi | 1 x 106 CFU/mL | Staphylococcus aureus(MRSA), COL | 1 x 106 CFU/mL |
| Haemophilus influenzatype A | 1 x 106 CFU/mL | Staphylococcusepidermidis (MRSE),ATCC 29887 | 1 x 106 CFU/mL |
| Hepatitis A virus | 1 x 106 TCID50/mL | Staphylococcussaprophyticus | 1 x 106 CFU/mL |
| Hepatitis B virus | 1 x 105 IU/mL | Streptococcus agalactiae | 1 x 106 CFU/mL |
| Hepatitis C virus | 1 x 105 IU/mL | Streptococcus mitis | 1 x 106 CFU/mL |
| HHV-6 (Z29 strain) | 1 x 105 copies/mL | Streptococcus mutans | 1 x 106 CFU/mL |
| HHV-6A | 1 x 105 copies/mL | Streptococcuspneumoniae | 1 x 106 CFU/mL |
| HHV-7 SB | 1 x 105 TCID50/mL | Streptococcus pyogenes,M1 | 1 x 106 CFU/mL |
| HHV-8 | 1 x 105 copies/mL | Streptococcus salivarius | 1 x 106 CFU/mL |
| HIV-1 IIIB | 1 x 105 TCID50/mL | Toxoplasma gondii | 1 x 106 tachyzoites/mL |
| HIV-2 NIHZ | 1 x 105 TCID50/mL | Trichomonas vaginalis | 1 x 106 trophozoites/mL |
| HSV-1 (McIntyre strain) | 1 x 105 TCID50/mL | Ureaplasma urealyticum | 1 x 106 CCU/mL |
| HSV-2 (G strain) | 1 x 105 TCID50/mL | ||
| Organism | Tested Concentration | Organism | Tested Concentration |
| Acholeplasma laidlawi(genomic DNA) | 1 x 106 copies/mL | Human genomic DNA | 1 x 106 copies/mL |
| Acinetobactercalcoaceticus | 1 x 106 CFU/mL | Human metapneumovirusA1 | 1 x 105 TCID50/mL |
| Acinetobacter Iwoffi | 1 x 106 CFU/mL | Human papilloma virus 18 | 1 x 105 copies/mL |
| Adenovirus 7A | 1 x 105 TCID50/mL | InfluenzaA/California/7/2009 | 1 x 105 TCID50/mL |
| Bacteroides fragilis | 1 x 106 CFU/mL | InfluenzaB/Florida/02/2006 | 1 x 105 TCID50/mL |
| Bordetella bronchiseptica | 1 x 106 CFU/mL | Klebsiella pneumoniae | 1 x 106 CFU/mL |
| Bordetella pertussis | 1 x 106 CFU/mL | Lactobacillus acidophilus | 1 x 106 CFU/mL |
| Borrelia burgdorferi(genomic DNA) | 1 x 106 copies/mL | Legionella pneumophila | 1 x 106 CFU/mL |
| Candida albicans | 1 x 106 CFU/mL | Measles virus | 1 x 105 TCID50/mL |
| Candida glabrata | 1 x 106 CFU/mL | Mobiluncus curtisii | 1 x 106 CFU/mL |
| Candida guilliermondii | 1 x 106 CFU/mL | Mobiluncus mulieris | 1 x 106 CFU/mL |
| Candida krusei | 1 x 106 CFU/mL | Moraxella cartarrhalis | 1 x 106 CFU/mL |
| Candida lusitaniae | 1 x 106 CFU/mL | Mumps virus | 1 x 105 TCID50/mL |
| Candida parapsilosis | 1 x 106 CFU/mL | Mycoplasma genitalium | 1 x 106 CCU/mL |
| Candida tropicalis | 1 x 106 CFU/mL | Mycoplasma hominis | 1 x 106 CCU/mL |
| Chlamydia trachomatis | 1 x 106 IFU/mL | Mycoplasma hyorhinis | 1 x 106 CCU/mL |
| Chlamydophilapneumoniae | 1 x 106 IFU/mL | Mycoplasma orale | 1 x 106 CCU/mL |
| Clostridium difficile | 1 x 106 CFU/mL | Mycoplasma pneumoniae | 1 x 106 CCU/mL |
| Clostridium perfringens | 1 x 106 CFU/mL | Mycoplasma salivarium | 1 x 106 CCU/mL |
| Clostridium sordellii | 1 x 106 CFU/mL | Neisseria gonorrhoeae | 1 x 106 CFU/mL |
| Coronavirus OC43 | 1 x 105 TCID50/mL | Neisseria meningitidis | 1 x 106 CFU/mL |
| Corynebacteriumdiphtheriae | 1 x 106 CFU/mL | Parainfluenza Type 1 | 1 x 105 TCID50/mL |
| Corynebacteriumgenitalium | 1 x 106 CFU/mL | Parainfluenza Type 2 | 1 x 105 TCID50/mL |
| Coxsackievirus B1 | 1 x 105 TCID50/mL | Parainfluenza Type 3 | 1 x 105 TCID50/mL |
| Coxsackievirus B4 | 1 x 105 TCID50/mL | Parainfluenza Type 4 | 1 x 105 TCID50/mL |
| Cytomegalovirus (AD169strain) | 1 x 105 TCID50/mL | Prevotella melaninogenica | 1 x 106 CFU/mL |
| Cytomegalovirus (Townestrain) | 1 x 105 TCID50/mL | Proteus mirabilis | 1 x 106 CFU/mL |
| Echovirus 11 | 1 x 105 TCID50/mL | Proteus vulgaris | 1 x 106 CFU/mL |
| Enterobacter cloacae | 1 x 106 CFU/mL | Pseudomonas aeruginosa | 1 x 106 CFU/mL |
| Enterococcus faecalisvanB | 1 x 106 CFU/mL | RSV A Long | 1 x 105 TCID50/mL |
| Organism | Tested Concentration | Organism | Tested Concentration |
| Enterococcus faecium | 1 x 106 CFU/mL | RSV B Washington | 1 x 105 TCID50/mL |
| Enterovirus 70 | 1 x 105 TCID50/mL | Rubella Virus | 1 x 105 TCID50/mL |
| Enterovirus 71 | 1 x 105 TCID50/mL | Salmonella enteritidis(genomic DNA) | 1 x 106 copies/mL |
| Epstein Barr Virus (B95-8strain) | 1 x 105 copies/mL | Salmonella typhimurium | 1 x 106 CFU/mL |
| Escherichia coli O15:H7 | 1 x 106 CFU/mL | Serratia marcescens | 1 x 106 CFU/mL |
| Fusobacterium nucleatum | 1 x 106 CFU/mL | Simian Virus type 40 | 1 x 105 TCID50/mL |
| Gardnerella vaginalis | 1 x 106 CFU/mL | Staphylococcus aureus(MRSA), ATCC 700699 | 1 x 106 CFU/mL |
| Haemophilus ducreyi | 1 x 106 CFU/mL | Staphylococcus aureus(MRSA), COL | 1 x 106 CFU/mL |
| Haemophilus influenzatype A | 1 x 106 CFU/mL | Staphylococcusepidermidis (MRSE),ATCC 29887 | 1 x 106 CFU/mL |
| Hepatitis A virus | 1 x 106 TCID50/mL | Staphylococcussaprophyticus | 1 x 106 CFU/mL |
| Hepatitis B virus | 1 x 105 IU/mL | Streptococcus agalactiae | 1 x 106 CFU/mL |
| Hepatitis C virus | 1 x 105 IU/mL | Streptococcus mitis | 1 x 106 CFU/mL |
| HHV-6 (Z29 strain) | 1 x 105 copies/mL | Streptococcus mutans | 1 x 106 CFU/mL |
| HHV-6A | 1 x 105 copies/mL | Streptococcuspneumoniae | 1 x 106 CFU/mL |
| HHV-7 SB | 1 x 105 TCID50/mL | Streptococcus pyogenes,M1 | 1 x 106 CFU/mL |
| HHV-8 | 1 x 105 copies/mL | Streptococcus salivarius | 1 x 106 CFU/mL |
| HIV-1 IIIB | 1 x 105 TCID50/mL | Toxoplasma gondii | 1 x 106 tachyzoites/mL |
| HIV-2 NIHZ | 1 x 105 TCID50/mL | Trichomonas vaginalis | 1 x 106 trophozoites/mL |
| HSV-1 (McIntyre strain) | 1 x 105 TCID50/mL | Ureaplasma urealyticum | 1 x 106 CCU/mL |
| HSV-2 (G strain) | 1 x 105 TCID50/mL |
{10}------------------------------------------------
Image /page/10/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 professional, suggesting a company involved in biotechnology or molecular diagnostics.
510(k) Summary
Simplexa™ VZV Swab Direct Simplexa™ VZV Positive Control Pack Page 9 of 15
Note: Bacteroides uredyticus, Hepatits D virus, Treponema whippler were tested using in silico NCB BLAST analysis due to unavailability of the organism. No cross-reactivity was found.
INHIBITION BY OTHER MICROORGANISMS
The Simplexa™ VZV Swab Direct assay was evaluated by testing the ability to identify VZV virus (Ellen and 9939 strains) when other potential inhibitory organisms are present. A panel of ninety-nine (99) potentialy inhibitory organisms were individually spiked into pooled cutaneous and mucocutaneous swab matrix containing a low concentration of VZV at approximately 2X LoD and tested in triplicate. Table 6 below references the microorqanisms and their respective tested concentration. No inhibition was observed for the detection of either VZV Ellen or 9939 strains as shown.
{11}------------------------------------------------
Image /page/11/Picture/0 description: The image contains 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 a dark blue, sans-serif font, with the word "Molecular" underneath in a green, sans-serif font.
510(k) Summary Simplexa™ VŽV Swab Direct
Simplexa™ VZV Positive Control Pack Page 10 of 15
Table 6. Simplexa™ VZV Swab Direct Microbial Inhibition
{12}------------------------------------------------
Image /page/12/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 company name, "DiaSorin," in a bold, dark blue font. Below "DiaSorin" is the word "Molecular" in a lighter green font.
510(k) S marv
Simplexa™ VZV Swa Direct Simplexa™ VZV Positive Con I Pack Page 11 of 15
Note: Bacteroides ureolyticus, Hepatitis D virus, Trepheryma whipplei were tested using in silico NCB IBLAST analysis due to unavailability of the organism. No interference was found.
INTERFERENCE
The performance of the Simplexa™ VZV Swab Direct assay was evaluated with potentially interfering substances. The tested concentrations of the potentially interfering endogenous and exogenous substances are indicated in the table below (Table 7). A total of forty-five (45) potential interfering substances were individually spiked into a pooled cutaneous and mucocutaneous swab matrix containing a low concentration of VZV at approximately 2X LoD and tested in triplicate. No interference was observed as presented in Table 7 for Ellen and 9939 Strains.
{13}------------------------------------------------
Image /page/13/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a stylized DNA helix in green and blue on the left, followed by the words "DiaSorin" in dark blue on the top line and "Molecular" in green on the bottom line. The logo is clean and modern, with a focus on the company's molecular diagnostics business.
510(k) Summary Simplexa™ VŽV Swab Direct
Simplexa™ VZV Positive Control Pack Page 12 of 15
Table 7. Simplexa™ VZV Swab Direct Interference
| Potentially InterferingSubstance. | VZV Strain | Active Ingredient | TestedConcentration | # Detected/# Tested |
|---|---|---|---|---|
| Abreva | 9936 | Docosanol 10% | 7% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Acetaminophen | 9936 | N/A | 7% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Acyclovir | 9936 | N/A | 10 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Albumin | 9936 | N/A | 10 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Balneol lotion | 9936 | Buffers, emulsifiers, PEG,water, mineral oil, lanolin oil,preservatives | 7% (v/v) | 3/3 |
| Ellen | 3/3 | |||
| Carmex | 9936 | Camphor, 1.7%; Menthol,0.7% | 10% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Casein | 9936 | N/A | 10 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Chlor-Trimeton | 9936 | Chlorpheniramine maleate | 5 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Cidofovir | 9936 | N/A | 2.5 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Clotrimazole VaginalCream | 9936 | Clotrimazole | 7% (w/v) | 3/3 |
| Ellen | 3.5% (w/v) | 3/3 | ||
| Cold-Eeze | 9936 | Zincum Gluconicum 2X | 10% (w/v) | 3/3 |
| 5% (w/v) | 3/3 | |||
| 9936 | 2.5% (w/v) | 3/3 | ||
| Cornstarch | 9936 | N/A | 1.25 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Denavir | 9936 | N/A | 2.5 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Potentially InterferingSubstance. | VZV Strain | Active Ingredient | TestedConcentration | # Detected/# Tested |
| Desitin | 9936 | Zinc Oxide, 40% | 7% (w/v) | 3/3 |
| Ellen | 3.5% (w/v) | 3/3 | ||
| Dextromethorphanhydrobromide(Robitussin-DM) | 9936 | Dextromethorphanhydrobromide | 10 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Douche | 9936 | N/A | 7% (v/v) | 3/3 |
| Ellen | 3/3 | |||
| Famciclovir | 9936 | N/A | 2.5 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Feces | 9936 | N/A | 2.5 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Foscarnet | 9936 | N/A | 1.25 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Glucose | 9936 | N/A | 11 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Gynol II contraceptivejelly | 9936 | Nonoxynol-9 (3%) | 7% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Human genomic DNA | 9936 | N/A | 20 $μ$ g/mL | 3/3 |
| Ellen | 3/3 | |||
| Immunoglobulin | 9936 | N/A | 10 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| KY Jelly | 9936 | N/A | 10 mg/mL | 3/3 |
| Ellen | 5% (w/v) | 3/3 | ||
| Lactate | 9936 | N/A | 2.2 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| Lanacane | 9936 | Benzethonium chloride,0.2%; Benzocaine, 20% | 7% (v/v) | 3/3 |
| Ellen | 3/3 | |||
| Lip-Clear Lysine | 9936 | Zinc Oxide, 1.2% | 7% (w/v) | 3/3 |
| Potentially InterferingSubstance. | VZV Strain | Active Ingredient | TestedConcentration | # Detected/# Tested |
| Ellen | 3.5% (w/v) | 3/3 | ||
| Miconazole 1 | 9936 | Miconazole nitrate, 26% | 10% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Miconazole 3 | 9936 | Miconazole nitrate, 2% | 10% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Monistat 1 insert | 9936 | Miconazole nitrate, 1200 mg | 7% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Monistat 3 cream | 9936 | Miconazole nitrate 2% | 7% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Mouthwash (Listerine) | 9936 | Eucalyptol, 0.092%; Menthol,0.042%; Methyl salicylate,0.060%; Thymol, 0.064% | 7% (v/v) | 3/3 |
| Ellen | 3/3 | |||
| Mucin | 9936 | N/A | 5% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Preparation H | 9936 | N/A | 10% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Releev | 9936 | N/A | 10% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Seminal Fluid | 9936 | N/A | 10% (v/v) | 3/3 |
| Ellen | 3/3 | |||
| Tioconazole 1 | 9936 | N/A | 10% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Toothpaste (Colgate) | 9936 | Sodium fluoride, 0.243% | 7% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Urine | 9936 | N/A | 10% (v/v) | 3/3 |
| Ellen | 3/3 | |||
| Vagisil creme | 9936 | Benzocaine (5%), Resorcinol(2%) | 7% (w/v) | 3/3 |
| Ellen | 3/3 | |||
| Valacyclovir | 9936 | N/A | 2.5 mg/mL | 3/3 |
| Potentially InterferingSubstance. | VZV Strain | Active Ingredient | TestedConcentration | # Detected/# Tested |
| Ellen | 3/3 | |||
| Valgancyclovir | 9936 | N/A | 2.5 mg/mL | 3/3 |
| Ellen | 3/3 | |||
| White blood cells | 9936 | N/A | 5.5x107 cells/mL | 3/3 |
| Ellen | 3/3 | |||
| Whole Blood in EDTA | 9936 | N/A | 10% (v/v) | 3/3 |
| Ellen | 3/3 | |||
| YeastGard suppositories | 9936 | Candida albicans 27X HPU(Candida albicans), Candidaparapsilosis 27X HPUS(Candida parapsilosis),Pulsatilla 27X HPUS(Meadow Anemone) | 7% (w/v) | 3/3 |
| Ellen | 3/3 |
{14}------------------------------------------------
Image /page/14/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 DNA graphic 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 Swab Direct
Simplexa™ VZV Positive Control Pack Page 13 of 15
{15}------------------------------------------------
Image /page/15/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, and below that, the word "Molecular" is written in a lighter green font.
510(k) Summary
Simplexa™ VZV Swab Direct
Simplexa™ VZV Positive Control Pack Page 14 of 15
{16}------------------------------------------------
Image /page/16/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo has a green and blue DNA helix on the left side. To the right of the helix is the text "DiaSorin" in blue, with the text "Molecular" in green below it.
510(k) Su
Simplexa™ VZV Sw Simplexa™ VZV Positive Cont
CARRY-OVER CONTAMINATION
The amplification carry-over for the Simplexa™ assays including the Simplexa™ VZV Swab Direct was assessed from the Simplexa™ Flu A/B & RSV Direct viral assay. The study can be applied to the Simplexa™ VZV Swab Direct assay as the study is not analyte specific. In the Simplexa™ Flu A/B & RSV Direct, the amplification carry-over study searched for the presence of contamination in negative samples adjacent to strong positive samples. The study was designed by alternately placing high positive and negative samples on each disc. No evidence of carry-over contamination was observed.
CONCLUSION
From the above analytical and comparative testing results of the Simplexa™ VZV Swab Direct assay it is concluded that it is substantially equivalent to the FDA cleared device Solana® HSV 1+2/VZV Assay (K162451).
§ 866.3309 Herpes virus nucleic acid-based cutaneous and mucocutaneous lesion panel.
(a)
Identification. A herpes virus nucleic acid-based cutaneous and mucocutaneous lesion panel is a qualitative in vitro diagnostic device intended for the simultaneous detection and differentiation of different herpes viruses in cutaneous and mucocutaneous lesion samples from symptomatic patients suspected of Herpetic infections. Negative results do not preclude infection and should not be used as the sole basis for treatment or other patient management decisions. The assay is not intended for use in cerebrospinal fluid samples.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include detailed documentation for the device description, including the device components, ancillary reagents required but not provided, and a detailed explanation of the methodology including primer design and selection.
(2) Premarket notification submissions must include detailed documentation from the following analytical and clinical performance studies: Analytical sensitivity (Limit of Detection), reactivity, inclusivity, precision, reproducibility, interference, cross reactivity, carry-over, and cross contamination.
(3) Premarket notification submissions must include detailed documentation of a clinical study using lesion samples in which Herpes Simplex Virus 1, Herpes Simplex Virus 2, or Varicella Zoster Virus DNA detection was requested. The study must compare the device performance to an appropriate well established reference method.
(4) 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.
(5) The device labeling must include a limitation statement that reads: “The device is not intended for use with cerebrospinal fluid or to aid in the diagnosis of HSV or VZV infections of the central nervous system (CNS).”
(6) Premarket notification submissions must include quality assurance protocols and a detailed documentation for device software, including, but not limited to, standalone software applications and hardware-based devices that incorporate software.
(7) The risk management activities performed as part of the manufacturer's 21 CFR 820.30 design controls must document an appropriate end user device training program that will be offered as part of efforts to mitigate the risk of failure to correctly operate the instrument.