K Number
K173498
Date Cleared
2018-08-13

(273 days)

Product Code
Regulation Number
866.3980
Reference & Predicate Devices
Predicate For
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP AuthorizedThirdpartyExpeditedreview
Intended Use

Simplexa™ Bordetella Direct MOL2750
The DiaSorin Molecular Simplexa™ Bordetella Direct MOL2750 assay is an in vitro diagnostic test intended for use on the LIAISON® MDX instrument for the qualitative detection and differentiation of Bordetella pertussis and Bordetella parapertussis nucleic acids from frozen nasopharyngeal (NPS) specimens from patients with signs and symptoms of Bordetella infection of the respiratory tract.
The Simplexa™ Bordetella Direct assay is performed on the LIAISON® MDX instrument and utilizes realtime PCR amplification to detect B. pertussis by targeting the IS481 insertional element of the B. pertussis genome and to detect B. parapertussis by targeting the IS1001 insertional element of the B. parapertussis genome. The IS481 insertional element can also be present in B. holmesii and B. bronchiseptica. Specimens collected from patients with respiratory infection caused by B. pertussis, B. holmesii or B. bronchiseptica may yield positive test results in IS481 assays. B. holmesii infection may cause clinical illness similar to B. pertussis, and mixed outbreaks involving both B. pertussis and B. holmesii infection have been reported. Additional testing should be performed if necessary to differentiate B. holmesii and B. pertussis. B. bronchiseptica is a rare cause of infection in humans. When clinical factors suggest that B. pertussis may not be the cause of respiratory infection, other clinically appropriate investigation(s) should be carried out in accordance with published guidelines.
Negative results for the Simplexa™ Bordetella Direct assay do not preclude Bordetella infection and positive results do not rule out co-infection with other respiratory pathogens. Results from the Simplexa™ Bordetella Direct assay should be used with other clinical findings and epidemiological information as an aid in diagnosis of Bordetella infection. Test results should not be used as the sole basis for treatment or other patient management decisions.
Simplexa™ Bordetella Positive Control Pack MOL 2760
The Simplexa™ Bordetella Positive Control Pack MOL2760 is intended to be used as a control with the Simplexa™ Bordetella Direct kit. This control is not intended for use with other assays or systems.

Device Description

The Simplexa™ Bordetella Direct assay system is a real-time PCR assay that enables the direct amplification, detection and differentiation of Bordetella pertussis and Bordetella parapertussis DNA from unprocessed nasopharyngeal swabs (NPS) without nucleic acid extraction. The system consists of the Simplexa™ Bordetella Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc and associated accessories.
In the Simplexa™ Bordetella Direct assay, primers and fluorescent probes are used together to amplify and detect Bordetella pertussis, Bordetella parapertussis and internal control targets. Insertion sequences IS481 and IS1001 are targeted to identify Bordetella pertussis and Bordetella parapertussis DNA respectively in the specimen. An internal control is used to detect PCR failure and/or inhibition.
The DiaSorin Molecular Simplexa™ Bordetella Direct kit contains sufficient reagents for 24 reactions. Upon receipt, store at -10 to -30ºC (do not use a frost-free freezer). Each vial contains sufficient material for a single reaction. Use within 30 minutes of thawing.

AI/ML Overview

Here's a breakdown of the acceptance criteria and the study proving the device meets those criteria, based on the provided text:

Acceptance Criteria and Reported Device Performance

Criteria CategoryAcceptance Criteria (Implicit)Reported Device Performance
Clinical PerformanceAcceptable Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) compared to a composite reference method.Bordetella pertussis (Prospective Samples):PPA: 91.9% (68/74); 95% CI: 83.4% to 96.2%NPA: 98.7% (1026/1039); 95% CI: 97.9% to 99.3%Bordetella parapertussis (Prospective Samples):PPA: 100.0% (13/13); 95% CI: 77.2% to 100.0%NPA: 99.6% (1096/1100); 95% CI: 99.1% to 99.9%Bordetella parapertussis (Contrived Samples):PPA: 100.0% (56/56); 95% CI: 93.6% to 100.0%NPA: 100.0% (56/56); 95% CI: 93.6% to 100.0%
ReproducibilityHigh agreement with expected results across different sites, days, and operators, with low variability (low %CV for Ct values).Overall Agreement with Expected Results: 100.0% (540/540) for all tested panel members (B. pertussis LP, MP; B. parapertussis LP, MP; negative; positive control). 95% CI: 99.3% to 100.0%. Avg. Ct %CV: Ranged from 0.7% to 3.9% across sites and targets.
Analytical Sensitivity (LoD)Detection of Bordetella species at low concentrations (lowest concentration detected as positive >95% of the time).B. pertussis (strains A639 & BAA-589): 14.7 CFU/mL and 20.9 CFU/mL respectively.B. parapertussis (strains A747 & E595): 347.3 CFU/mL and 239.0 CFU/mL respectively.
Analytical ReactivityDetection of various Bordetella strains.Twelve B. pertussis strains detected at or below 80 CFU/mL (3/3 detection for all). Six B. parapertussis strains detected at or below 590 CFU/mL (3/3 detection for all). In silico BLAST analysis predicted detection of 294 additional B. pertussis and 5 additional B. parapertussis strains.
Analytical Specificity (Cross-Reactivity)No detection of closely related organisms, organisms causing similar symptoms, or normal flora (except expected cross-reactivity).No cross-reactivity observed with 96 out of 97 tested organisms. Exception: Bordetella holmesii showed 100% detection (8/8) for the B. pertussis (IS481) target, which was expected due to the presence of the IS481 element in B. holmesii.
InterferenceNo interference from common substances found in nasopharynx.No evidence of interference caused by 16 tested substances (e.g., Albuterol, Blood, Mucin) on the detection of B. pertussis or B. parapertussis at 2-4 X LoD. (100% detection for all tested interferences except initial Rifampicin for B. parapertussis, which was resolved with additional replicates)
Competitive InterferenceNo interference between detection of B. pertussis and B. parapertussis when one is present at high concentration and the other at low.Low level of B. pertussis was detected in the presence of a high level of B. parapertussis (3/3 detection for both). Low level of B. parapertussis was detected in the presence of a high level of B. pertussis (3/3 detection for both).
Inhibition by Other MicroorganismsNo inhibition of Bordetella detection when other microorganisms are present.No inhibitory effects observed for B. pertussis and B. parapertussis (at 2X LoD) when spiked with 97 different potentially inhibitory organisms. 100% detection for both targets across all tested organisms including baseline (total of 45 tests for baseline and 3 tests per organism).
Carry-over ContaminationNo carry-over contamination between high positive and negative samples."No evidence of carry-over contamination was observed."

Study Details from the Provided Text:

2. Sample size used for the test set and the data provenance:

  • Test Set Sample Size (Clinical Performance/Method Comparison):
    • 1113 evaluable prospectively collected frozen nasopharyngeal swab (NPS) samples.
    • An additional 112 samples: 56 contrived Bordetella parapertussis samples (at 2-50 X LoD) and 56 negative samples, randomized together.
  • Data Provenance:
    • Country of Origin: Not explicitly stated, but samples were "prospectively collected and frozen from five (5) geographically diverse sites" (Page 7). This often implies within the USA for FDA submissions, but not definitively confirmed.
    • Retrospective or Prospective: Primarily prospective.
      • "One thousand one hundred and forty-two (1142) samples were prospectively collected and frozen..." (Page 7).
      • "The Bordetella pertussis prospectively banked frozen sample results are shown in Table 1." (Page 7).
      • "The Bordetella parapertussis prospectively banked frozen sample results are shown in Table 2 and the Bordetella parapertussis contrived sample results are shown in Table 3." (Page 7).

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts:

  • The text does not specify the number or qualifications of experts directly establishing ground truth. Instead, it describes a "composite reference method."

4. Adjudication method for the test set:

  • The ground truth was established using a "composite reference method [which] consisted of two well-characterized real-time PCR assays followed by confirmation of positive PCR amplification products with bi-directional sequencing, per target."
  • "Samples were characterized as positive if one or both composite reference methods were positive and confirmed by bi-directional sequencing. Samples were characterized as negative if both composite reference methods were negative." (Page 7).
  • This suggests a form of consensus/confirmation, but not a 2+1 or 3+1 expert adjudication in the typical human-in-the-loop sense for imaging. It's an algorithmic/laboratory "adjudication" against established molecular methods.

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:

  • No, an MRMC comparative effectiveness study was not done. This device is an in vitro diagnostic (IVD) PCR assay, not an AI-assisted imaging device. Its performance is evaluated directly against a reference method, not in comparison to human readers.

6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done:

  • Yes, the primary clinical performance ("Method Comparison") and all analytical studies (Reproducibility, LoD, Reactivity, Specificity, Interference, etc.) represent the standalone performance of the Simplexa™ Bordetella Direct assay. It's an automated molecular diagnostic assay, designed to operate without human interpretation of the primary signal for diagnosis.

7. The type of ground truth used:

  • The ground truth was established using a composite reference method consisting of:
    • Two "well-characterized real-time PCR assays."
    • "Confirmation of positive PCR amplification products with bi-directional sequencing, per target." (Page 7).

8. The sample size for the training set:

  • The document describes performance studies for a finalized device. It does not provide information about a separate "training set" in the context of machine learning, because this is a PCR assay with defined primers and probes, not a machine learning model that requires training data in that sense. The analytical and clinical studies serve to validate the assay's performance.

9. How the ground truth for the training set was established:

  • Not applicable, as this is not a machine learning device with a distinct "training set" ground truth. The development of the assay (designing primers, probes, optimizing conditions) is an engineering process, not a data-driven model training process.

{0}------------------------------------------------

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, with flowing lines representing hair or clothing.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

August 13, 2018

DiaSorin Molecular LLC Sharon Young Senior Regulatory Affairs Specialist 11331 Valley View Street Cypress, CA 90630

Re: K173498

Trade/Device Name: Simplexa Bordetella Direct MOL2750. Simplexa Bordetella Positive Control Pack MOL2760 Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: Class II Product Code: OZZ Dated: November 16, 2017 Received: November 17, 2017

Dear Ms. 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. 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 801 and Part 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the

{1}------------------------------------------------

Page 2 - Sharon Young

electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and Part 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. 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 the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely,

Kristian M. Roth -S

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

{2}------------------------------------------------

Indications for Use

510(k) Number (if known)

Device Name

Simplexa™ Bordetella Direct MOL2750 and Simplexa™ Bordetella Positive Control Pack MOL 2760

Indications for Use (Describe)

Simplexa™ Bordetella Direct MOL2750

The DiaSorin Molecular Simplexa™ Bordetella Direct MOL2750 assay is an in vitro diagnostic test intended for use on the LIAISON® MDX instrument for the qualitative detection and differentiation of Bordetella parapertussis nucleic acids from frozen nasopharyngeal (NPS) specimens with signs and symptoms of Bordetella infection of the respiratory tract.

The Simplexa™ Bordetella Direct MOL2750 assay is performed on the LIAISON® MDX instrument and utilizes realtime PCR amplification to detect B. pertussis by targeting the IS481 insertional element of the B. pertussis genome and to detect B. parapertussis by targeting the IS1001 insertional element of the B. parapertussis genome. The IS481 insertional element can also be present in B. holmesii and B. bronchiseptica. Specimens with respiratory infection caused by B. pertussis, B. holmesii or B. bronchiseptica may yield positive test results in IS481 assays. B. holmesii infection may cause clinical illness similar to B. pertussis, and mixed outbreaks involving both B. pertussis and B. holmesii infection have been reported. Additional testing should be performed if necessary to differentiate B. holmesii and B. pertussis. B. bronchiseptica is a rare cause of infection in humans. When clinical factors suggest that B. pertussis may not be the cause of respiratory infection, other clinically appropriate investigation(s) should be carried out in accordance with published guidelines.

Negative results for the Simplexa™ Bordetella Direct MOL2750 assay do not prectude Bordetella infection and positive results do not rule out co-infection with other respiratory pathogens. Results from the SimplexaTM Direct MOL2750 assay should be used with other clinical findings and epidemiological information as an aid in diagnosis of Bordetella infection. Test results should not be used as the sole basis for treatment or other patient management decisions.

Simplexa™ Bordetella Positive Control Pack MOL 2760

The Simplexa™ Bordetella Positive Control Pack MOL2760 is intended to be used as a control with the Simplexa™ Bordetella Direct kit. This control is not intended for use with other assays or systems.

Type of Use (Select one or both, as applicable)
---------------------------------------------------

X | Prescription Use (Part 21 CFR 801 Subpart D)

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

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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

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

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

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

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

{3}------------------------------------------------

Image /page/3/Picture/1 description: The image shows the logo for DiaSorin Molecular. The logo consists of a DNA helix graphic in green and blue on the left, with the text "DiaSorin" in dark blue on the top right. Below "DiaSorin" is the word "Molecular" in green.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 1 of 17

ApplicantDiaSorin Molecular LLC.11331 Valley View StreetCypress, California 90630USA
Establishment Registration No.2023365
Contact PersonSharon Youngtel 562.240.6680fax 562.240.6529Sharon.Young@DiaSorin.com
Summary DateAugust 7, 2018
Proprietary NameSimplexa™ Bordetella Direct and Simplexa™ Bordetella PositiveControl Pack
Generic NameBordetella nucleic acid
ClassificationClass II
Predicate DevicesARIES® Bordetella Assay for use with the ARIES® Systems

Intended Use

Simplexa™ Bordetella Direct

The DiaSorin Molecular Simplexa™ Bordetella Direct assay is an in vitro diagnostic test intended for use on the LIAISON® MDX instrument for the qualitative detection and differentiation of Bordetella perfussis and Bordetella parapertussis nucleic acids from frozen nasopharyngeal (NPS) specimens from patients with signs and symptoms of Bordetella infection of the respiratory tract.

The Simplexa™ Bordetella Direct assay is performed on the LIAISON® MDX instrument and utilizes realtime PCR amplification to detect B. pertussis by targeting the IS481 insertional element of the B. pertussis genome and to detect B. parapertussis by targeting the IS1001 insertional element of the B. parapertussis genome. The IS481 insertional element can also be present in B. holmesii and B. bronchiseptica. Specimens collected from patients with respiratory infection caused by B. pertussis, B. holmesii or B. bronchiseptica may vield positive test results in IS481 assays. B. holmesii infection may cause clinical illness similar to B. pertussis, and mixed outbreaks involving both B. pertussis and B. holmesii infection have been reported. Additional testing should be performed if necessary to differentiate B. holmesii and B. pertussis. B. bronchiseptica is a rare cause of infection in humans. When clinical factors suggest that B. pertussis may not be the cause of respiratory infection, other clinically appropriate investigation(s) should be carried out in accordance with published guidelines.

Negative results for the Simplexa™ Bordetella Direct assay do not preclude Bordetella infection and positive results do not rule out co-infection with other respiratory pathogens. Results from the Simplexa™ Bordetella Direct assay should be used with other clinical findings and epidemiological information as an aid in diagnosis of Bordetella infection. Test results should not be used as the sole basis for treatment or other patient management decisions.

Simplexa™ Bordetella Positive Control Pack

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

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

Device Description

The Simplexa™ Bordetella Direct assay system is a real-time PCR assay that enables the direct amplification, detection and differentiation of Bordetella pertussis and Bordetella parapertussis DNA from unprocessed nasopharyngeal swabs (NPS) without nucleic acid extraction. The system consists of the

{4}------------------------------------------------

Image /page/4/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 is the company name, "DiaSorin," in a dark blue, serif font. Below "DiaSorin" is the word "Molecular" in a lighter green sans-serif font.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 2 of 17

Simplexa™ Bordetella Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc and associated accessories.

In the Simplexa™ Bordetella Direct assay, primers and fluorescent probes are used together to amplify and detect Bordetella pertussis, Bordetella parapertussis and internal control targets. Insertion sequences IS481 and IS1001 are targeted to identify Bordetella pertussis and Bordetella parapertussis DNA respectively in the specimen. An internal control is used to detect PCR failure and/or inhibition.

The DiaSorin Molecular Simplexa™ Bordetella Direct kit contains sufficient reagents for 24 reactions. Upon receipt, store at -10 to -30ºC (do not use a frost-free freezer). Each vial contains sufficient material for a single reaction. Use within 30 minutes of thawing.

Kit Description

Component NameREFEC SYMBOLON LABELAbbreviatedNameCapColorNumberof VialsReactions Volumeper Vial/Kit per Vial
Simplexa™Bordetella DirectReaction MixMOL2751REAGRMBrown241/2450 ul

Component Description

Kit ComponentContents
DNA polymerase, buffer, dNTPs, Internal Control template DNA, dye-labeled fluorescent probes and primersspecific for detection of Bordetella pertussis , Bordetella parapertussis and DNA Internal Control
Simplexa™Bordetella DirectReaction Mix (RM)TargetProbeFluorophore(Dye)Excitation (nm)Emission (nm)TargetedGene
Bordetella pertussisFAM495520IS481
BordetellaparapertussisCFR610590610IS1001
DNA Internal ControlQ670644670DNA IC
Simplexa™Bordetella KitBarcode CardAssay specific parameters

MATERIALS SUPPLIED SEPARATELY

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

{5}------------------------------------------------

Image /page/5/Picture/0 description: The image shows the logo for DiaSorin Molecular. The word "DiaSorin" is in a dark blue font, and the word "Molecular" is in a light green font. To the left of the text is a green and blue DNA strand graphic.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 3 of 17

Predicate Device Information

ItemDevice
NameSimplexa™ Bordetella Direct
REF MOL2750
ARIES® Bordetella Assay for use withAnd
the ARIES® Systems (K163626)Simplexa™ Bordetella Positive Control
Pack REF MOL2760
Intended UseThe ARIES® Bordetella Assay is a real-time polymerase chain reaction (PCR)based qualitative in vitro diagnostic testfor the direct detection and identificationof Bordetella pertussis (B. pertussis) andBordetella parapertussis (B.parapertussis) nucleic acid innasopharyngeal swab (NPS) specimensobtained from individuals suspected ofhaving a respiratory tract infectionattributable to B. pertussis or B.parapertussis. The ARIES® BordetellaAssay targets the B. pertussis toxinpromoter and the B. parapertussisIS1001 insertion element in thegenomes. When clinical factors suggestthat B. pertussis or B. parapertussis maynot be the cause of respiratory infection,other clinically appropriateinvestigation(s) should be carried out inaccordance with published guidelines.Negative results for the ARIES®Bordetella Assay do not preclude B.pertussis or B. parapertussis infectionand positive results do not rule out co-infections with other respiratorypathogens. The direct detection andidentification of B. pertussis and B.parapertussis nucleic acids fromsymptomatic patients aids in thediagnosis of B. pertussis and B.parapertussis respiratory infection inconjunction with other clinical findingsand epidemiological information. TheARIES® Bordetella Assay is indicated foruse with the ARIES® Systems.Simplexa™ Bordetella DirectThe DiaSorin Molecular Simplexa™Bordetella Direct assay is an in vitrodiagnostic test intended for use on theLIAISON® MDX instrument for the qualitativedetection and differentiation of Bordetellapertussis and Bordetella parapertussisnucleic acids from frozen nasopharyngeal(NPS) specimens from patients with signsand symptoms of Bordetella infection of therespiratory tract.The Simplexa™ Bordetella Direct assay isperformed on the LIAISON® MDX instrumentand utilizes real-time PCR amplification todetect B. pertussis by targeting the IS481insertional element of the B. pertussisgenome and to detect B. parapertussis bytargeting the IS1001 insertional element ofthe B. parapertussis genome. The IS481insertional element can also be present in B.holmesii and B. bronchiseptica. Specimenscollected from patients with respiratoryinfection caused by B. pertussis, B. holmesiior B. bronchiseptica may yield positive testresults in IS481 assays. B. holmesii infectionmay cause clinical illness similar to B.pertussis, and mixed outbreaks involvingboth B. pertussis and B. holmesii infectionhave been reported. Additional testingshould be performed if necessary todifferentiate B. holmesii and B. pertussis. B.bronchiseptica is a rare cause of infection inhumans. When clinical factors suggest thatB. pertussis may not be the cause ofrespiratory infection, other clinicallyappropriate investigation(s) should be carriedout in accordance with published guidelines.Negative results for the Simplexa TMBordetella Direct assay do not precludeBordetella infection and positive results donot rule out co-infection with other respiratorypathogens. Results from the Simplexa™
ItemDevice
NameARIES® Bordetella Assay for use withthe ARIES® Systems (K163626)Simplexa™ Bordetella DirectREF MOL2750AndSimplexa™ Bordetella Positive ControlPack REF MOL2760
Bordetella Direct assay should be used withother clinical findings and epidemiologicalinformation as an aid in diagnosis ofBordetella infection. Test results should notbe used as the sole basis for treatment orother patient management decisions.Simplexa™ Bordetella Positive Control Pack
The Simplexa™ Bordetella Positive ControlPack is intended to be used as a control withthe Simplexa™ Bordetella Direct kit.This control is not intended for use with otherassays or systems.
Assay TargetsB. pertussis Target B. pertussis and B. parapertussis toxin promoter (ptxA-pr),B. parapertussis Target IS1001 insertionelement .The Simplexa™ Bordetella Direct assaydetects the following multi-copy insertionsequence targets: Bordetella pertussis targetIS481 and Bordetella parapertussis targetIS1001.
Sample TypesNasopharyngeal swabs in UTM™, M5®,M6™ and ESwab.Nasopharyngeal swabs in Remel M4, RemelM4RT, Remel M5, Remel M6, UTM, VTMand Liquid Aimes (ESwab).
InstrumentARIES® System, ARIES® M1 System.LIAISON® MDX.
Extraction MethodsNone.None.
Assay MethodologyReal-time system for detecting thepresence / absence of Bordetella pertussis and Bordetella parapertussisDNA in clinical specimens.PCR-based system for detecting thepresence / absence of Bordetella pertussisand Bordetella parapertussis DNA in clinicalspecimens.
Detection TechniquesDifferent fluorescent reporter dyes foreach target and melt analysis.Multiplex assay using different reporter dyesfor each target.
ItemDevice
NameARIES® Bordetella Assay for use withthe ARIES® Systems (K163626)Simplexa™ Bordetella DirectREF MOL2750AndSimplexa™ Bordetella Positive ControlPack REF MOL2760
OpticalDetectionFluorescence Emissions and Detection.Fluorescence.
Time to resultLess than 2 hours.Approximately 1 hour
TestInterpretationAutomated test interpretation and reportgeneration.Automated test interpretation and reportgeneration.

{6}------------------------------------------------

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

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 4 of 17

{7}------------------------------------------------

Image /page/7/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, the word "DiaSorin" is written in a dark blue sans-serif font. Below "DiaSorin", the word "Molecular" is written in a lighter green sans-serif font.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 5 of 17

METHOD COMPARISON

One thousand one hundred and forty-two (1142) samples were prospectively collected and frozen from five (5) geographically diverse sites between January 2017, from patients with signs and symptoms of Bordetella infections. Of the one thousand one hundred and forty-two (1142) samples, one hundred and thirteen (1113) samples were evaluable on Simplexa™ Bordetella Direct and a composite reference method. In addition, fifty-six (56) samples were contrived for Bordetella parapertussis at various concentrations across the clinical range of the assay (2-50 X LoD) and randomized among fifty-six (56) negatives for a total of one hundred and twelve (112) additional samples, which were evaluable on Simplexa™ Direct and a composite reference method. The composite reference method consisted of two well-characterized real-time PCR assays followed by confirmation of positive PCR amplification products with bi-directional sequencing, per target. Samples were characterized as positive if one or both composite reference methods were positive and confirmed by bi-directional sequencing. Samples were characterized as negative if both composite reference methods were negative. Samples were tested on Simplexa™ Bordetella Direct at the collection sites and the composite reference method was performed at DiaSorin Molecular. The Bordetella pertussis prospectively banked frozen sample results are shown in Table 1. The Bordetella parapertussis prospectively banked frozen sample results are shown in Table 2 and the Bordetella parapertussis contrived sample results are shown in Table 3.

Table 1. Simplexa™ Bordetella Direct Bordetella pertussis Results Versus PCR/Bi-Directional Sequencing Method Prospectively Banked Frozen Samples

Simplexa™Bordetella DirectB. pertussisPCR/Bi-directional Sequencing
DetectedNot DetectedTotal
Detected681381
Not Detected610261032
Total7410391113
%PPA91.9%(68/74)95% CI: 83.4% to 96.2%%NPA98.7%(1026/1039)95% CI: 97.9% to 99.3%

{8}------------------------------------------------

Image /page/8/Picture/1 description: The image shows the logo for DiaSorin Molecular. The logo consists of a DNA helix graphic in shades of green and blue on the left, followed by the text "DiaSorin" in dark blue on the top line. Below that, the word "Molecular" is written in green.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 6 of 17

Table 2. Simplexa™ Bordetella Direct Bordetella parapertussis Results versus PCR/Bi-Directional Sequencing Method Prospectively Collected Frozen Samples

Simplexa™PCR/Bi-directional Sequencing
Bordetella DirectB. parapertussisDetectedNot DetectedTotal
Detected13417
Not DetectedO10961096
Total1311001113
%PPA100.0%(13/13)95% Cl: 77.2% to 100.0%%NPA99.6%(1096/1100)95% CI: 99.1% to 99.9%

Table 3. Simplexa™ Bordetella Direct Bordetella parapertussis results versus PCR/Bi-Directional Sequencing Method Contrived Frozen Samples

Simplexa™Bordetella DirectB. parapertussisPCR/Bi-directional Sequencing
DetectedNot DetectedTotal
Detected56056
Not Detected05656
Total5656112
%PPA100.0%(56/56)95% CI: 93.6% to 100.0%%NPA100.0%(56/56)95% CI: 93.6% to 100.0%

REPRODUCIBILITY

Three (3) investigative laboratory testing sites assessed the device's inter-day and interlintraassay reproducibility. Each of the laboratories tested a panel of six (6) members that included contrived Bordetella pertussis A639, and Bordetella parapertussis A747 samples at the following concentrations; low positive (LP) approximately 1-2 X LoD and a medium positive sample (MP) approximately 3-4 X LoD. A positive and negative control was also included in the panel. Each sample panel member was tested in triplicate per run for two (2) runs per day for five (5) non-consecutive days per site. Two (2) runs per day were each performed by a different operator. In total ninety (90) replicates X 2 runs X 5 days X 3 sites) were tested for each sample panel member. Combined results for all sites are presented in Table 4. The data in Table 4 show that the Simplexa™ Bordetella Direct is 100% in agreement with the expected results demonstrating the reproducibility of the assay for both Bordetella pertussis and Bordetella parapertussis targets.

{9}------------------------------------------------

Image /page/9/Picture/0 description: The image contains a sequence of alphanumeric characters. The characters are 'K173498' and are presented in a bold, sans-serif font. The characters are black against a white background.

Image /page/9/Picture/1 description: The image shows the logo for DiaSorin Molecular. The logo features a green and blue DNA helix on the left side. To the right of the helix, the words "DiaSorin" are written in blue, and below that, the word "Molecular" is written in green.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 7 of 17

Site 1Site 2Site 3Overall
Sample PanelMember%AgreementwithExpectedResultsAvg.Ct%CV%AgreementwithExpectedResultsAvg.Ct%CV%AgreementwithExpectedResultsAvg.Ct%CVTotal %95% Cl
BordetellapertussisA639 - LP(FAM)100.0%(30/30)33.93.4100.0%(30/30)34.23.3100.0%(30/30)34.92.6100.0%(90/90)95.9%to100.0%
BordetellapertussisA639 - MP(FAM)100.0%(30/30)32.92.5100.0%(30/30)33.82.9100.0%(30/30)33.92.7100.0%(90/90)95.9%to100.0%
Bordetellapara-pertussisA747 - LP(CFR 610)100.0%(30/30)33.93.9100.0%(30/30)34.32.7100.0%(30/30)35.43.5100.0%(90/90)95.9%to100.0%
Bordetellapara-pertussisA747 - MP(CFR 610)100.0%(30/30)33.02.7100.0%(30/30)33.52.6100.0%(30/30)33.72.0100.0%(90/90)95.9%to100.0%
Nativenegativenaso-pharyngealswab (UTM)100.0%(30/30)N/AN/A100.0%(30/30)N/AN/A100.0%(30/30)N/AN/A100.0%(90/90)95.9%to100.0%
PositiveControl(FAM)100.0%(30/30)24.00.8100.0%(30/30)22.01.1100.0%(30/30)23.30.7100.0%(90/90)95.9%to100.0%
PositiveControl(CFR 610)100.0%(30/30)23.60.9100.0%(30/30)20.81.0100.0%(30/30)22.40.7100.0%(90/90)95.9%to100.0%
TotalAgreement100.0% (180/180)100.0% (180/180)100.0% (180/180)100.0% (540/540)99.3% to 100.0%

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

The Limit of Detection (LoD) was determined for the Simplexa™ Bordetella Direct assay using quantified stocks of two (2) strains of Bordetella pertussis (A639 & BAA-589) and Bordetella parapertussis (A747 & E595) serially diluted into native nasopharyngeal swab matrix in Universal Transport Media (UTM). LoD is the lowest concentration that could be detected as positive > 95% of the time. The LoD concentrations for the Bordetella strains are summarized in Table 5.

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

Image /page/10/Picture/1 description: The image shows the logo for DiaSorin Molecular. The logo consists of a stylized DNA double helix on the left, rendered in shades of green and blue. To the right of the helix is the text "DiaSorin" in a dark blue, sans-serif font. Below "DiaSorin" is the word "Molecular" in a lighter green, sans-serif font, aligned with the right edge of "DiaSorin."

510(k) Summary Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 8 of 17

Bordetella speciesBordetella strainLoD Concentration (CFU/mL)
Bordetella pertussisA63914.7
Bordetella pertussisBAA-58920.9
Bordetella parapertussisA747347.3
Bordetella parapertussisE595239.0

Table 5. Simplexa™ Bordetella Direct Limit of Detection

ANALYTICAL REACTIVITY / CROSS REACTIVITY

Analytical Reactivity

Analytical Reactivity for Simplexa™ Bordetella Direct was assessed using eighteen (18) Bordetella strains including twelve (12) Bordetella pertussis strains and six (6) Bordetella parapertussis strains that were not tested as a part of the Limit of Detection (LoD) study. All eighteen (18) strains were detected as positive for Bordetella pertussis at or below 80 CFU/mL or for Bordetella parapertussis at or below 590 CFU/mL. In addition to the strains that were tested, in silico BLAST analysis demonstrated that the assay should detect at least two hundred and ninety-four (294) additional Bordetella pertussis and five (5) additional Bordetella parapertussis strains. The results of the testing are summarized in Table 6 for Bordetella pertussis and in Table 7 for Bordetella parapertussis.

Table 6. Simplexa™ Bordetella Direct Analytical Reactivity - Bordetella pertussis
Bordetella StrainConcentrationBordetella pertussis(IS481)Result (# detected/# tested)
Bordetella pertussis BAA-133535.6 CFU/mL3/3
Bordetella pertussis ATCC 846735.6 CFU/mL3/3
Bordetella pertussis ATCC 930679.7 CFU/mL3/3
Bordetella pertussis ATCC 1274235.6 CFU/mL3/3
Bordetella pertussis ATCC 5144535.6 CFU/mL3/3
Bordetella pertussis ATCC 5389479.7 CFU/mL3/3
Bordetella pertussis ATCC 847835.6 CFU/mL3/3
Bordetella pertussis ATCC 1274335.6 CFU/mL3/3
Bordetella pertussis ATCC 934035.6 CFU/mL3/3
Bordetella pertussis ATCC 979735.6 CFU/mL3/3
Bordetella pertussis ATCC 1038035.6 CFU/mL3/3
Bordetella pertussis E43135.6 CFU/mL3/3

{11}------------------------------------------------

Image /page/11/Picture/0 description: The image shows the alphanumeric string 'K173498' in a bold, sans-serif font. The characters are black against a white background. The string appears to be a code or identifier.

Image /page/11/Picture/1 description: The image shows the logo for DiaSorin Molecular. The logo consists of a green and blue DNA helix graphic 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 expertise in molecular diagnostics.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 9 of 17

Table 7. Simplexa™ Bordetella Direct Analytical Reactivity - Bordetella parapertussis

Bordetella StrainConcentrationBordetella parapertussis(IS1001)Result (# detected/# tested)
Bordetella parapertussis ATCC 15311586.3 CFU/mL3/3
Bordetella parapertussis ATCC 15237586.3 CFU/mL3/3
Bordetella parapertussis ATCC 15989586.3 CFU/mL3/3
Bordetella parapertussis BAA-587586.3 CFU/mL3/3
Bordetella parapertussis C510586.3 CFU/mL3/3
Bordetella parapertussis E838586.3 CFU/mL3/3

Cross Reactivity (Analytical Specificity)

Analytical specificity was evaluated for the Simplexa™ Bordetella Direct by testing cross reactivity to organisms that are closely related, or cause similar clinical symptoms, or are present as normal flora in the nasopharynx. Negative specimens were spiked with potentially cross reactive organisms at the concentrations indicated in Table 8. The spiked specimens were examined for reactivity with Simplexa™ Bordetella Direct. Ninety-seven (97) organisms were tested. No cross reactivity was found with the exception of Bordetella holmseli which was expected due to the presence of the IS481 element in Bordetella holmesii. The results are shown in Table 8.

Table 8. Simplexa™ Bordetella Direct Cross Reactivity

Cross ReactantConcentrationBordetella pertussis(IS481)% Detection(# Detected/# Tested)Bordetella parapertussis(IS1001)% Detection(# Detected/# Tested)
Acinetobacter baumannii1 x 106 CFU/mL0% (0/3)0% (0/3)
Acinetobacter Iwoffi1 x 106 CFU/mL0% (0/3)0% (0/3)
Adenovirus 11 x 105 TCID50/mL0% (0/3)0% (0/3)
Adenovirus 311 x 105 TCID50/mL0% (0/3)0% (0/3)
Arcanobacterium haemolyticum1 x 106 CFU/mL0% (0/3)0% (0/3)
Bacillus cereus1 x 106 CFU/mL0% (0/3)0% (0/3)
Bacteroides fragilis1 x 106 CFU/mL0% (0/3)0% (0/3)
Bordetella avium1 x 106 CFU/mL0% (0/3)0% (0/3)
Bordetella bronchiseptica RB501 x 106 CFU/mL0% (0/3)0% (0/3)
Bordetella hinzi1 x 106 CFU/mL0% (0/3)0% (0/3)
Bordetella holmseii F0611 x 106 CFU/mL100% (8/8)0% (0/3)
Bordetella petri1 x 106 CFU/mL0% (0/3)0% (0/3)
Bordetella trematum1 x 106 CFU/mL0% (0/3)0% (0/3)
Burkholderia cenocepacia1 x 106 CFU/mL0% (0/3)0% (0/3)
Burkholderia cepacia1 x 106 CFU/mL0% (0/3)0% (0/3)
Burkholderia multivorans1 x 106 CFU/mL0% (0/3)0% (0/3)
Burkholderia thailandensis1 x 106 CFU/mL0% (0/3)0% (0/3)
Cross ReactantConcentrationBordetella pertussis(IS481)% Detection(# Detected/# Tested)Bordetella parapertussis(IS1001)% Detection(# Detected/# Tested)
Candida albicans1 x 106 CFU/mL0% (0/3)0% (0/3)
Candida glabrata1 x 106 CFU/mL0% (0/3)0% (0/3)
Chlamydia pneumoniae1 x 106 IFU/mL0% (0/3)0% (0/3)
Chlamydia trachmomatis1 x 106 IFU/mL0% (0/3)0% (0/3)
Citrobacter freundii1 x 106 CFU/mL0% (0/3)0% (0/3)
Clostridium difficile1 x 106 CFU/mL0% (0/3)0% (0/3)
Coronavirus 229E1 x 105 TCID50/mL0% (0/3)0% (0/3)
Coronavirus NL63*1 x 104 TCID50/mL0% (0/3)0% (0/3)
Coronavirus OC431 x 105 TCID50/mL0% (0/3)0% (0/3)
Corynebacterium diptheriae1 x 106 CFU/mL0% (0/3)0% (0/3)
Coxsackievirus A161 x 105 TCID50/mL0% (0/3)0% (0/3)
Coxsackievirus B41 x 105 TCID50/mL0% (0/3)0% (0/3)
Cytomegalovirus1 x 105 TCID50/mL0% (0/3)0% (0/3)
Echovirus 61 x 105 TCID50/mL0% (0/3)0% (0/3)
Echovirus 71 x 105 TCID50/mL0% (0/3)0% (0/3)
Echovirus 91 x 105 TCID50/mL0% (0/3)0% (0/3)
Echovirus 111 x 105 TCID50/mL0% (0/3)0% (0/3)
Enterobacter aerogenes Z0521 x 106 CFU/mL0% (0/3)0% (0/3)
Enterobacter cloacae1 x 106 CFU/mL0% (0/3)0% (0/3)
Enterococcus faecalis vanB1 x 106 CFU/mL0% (0/3)0% (0/3)
Enterovirus 701 x 105 TCID50/mL0% (0/3)0% (0/3)
Enterovirus 711 x 105 TCID50/mL0% (0/3)0% (0/3)
Epstein-Barr Virus1 x 105 copies/mL0% (0/3)0% (0/3)
Escherichia coli1 x 106 CFU/mL0% (0/3)0% (0/3)
Fusobacterium necrophorum1 x 106 CFU/mL0% (0/3)0% (0/3)
Haemophilus influenzae1 x 106 CFU/mL0% (0/3)0% (0/3)
Haemophilus parainfluenzae1 x 106 CFU/mL0% (0/3)0% (0/3)
HSV-1 (MacIntyre)1 x 105 TCID50/mL0% (0/3)0% (0/3)
HSV-2 (G)1 x 105 TCID50/mL0% (0/3)0% (0/3)
Influenza A/Swine/lowa/15/30H1N1*1 x 104 TCID50/mL0% (0/3)0% (0/3)
Influenza B/Malaysia/2506/04*1 x 104 TCID50/mL0% (0/3)0% (0/3)
Klebsiella oxytoca1 x 106 CFU/mL0% (0/3)0% (0/3)
Klebsiella pneumoniae1 x 106 CFU/mL0% (0/3)0% (0/3)
Lactobacillus acidophilus1 x 106 CFU/mL0% (0/3)0% (0/3)
Lactobacillus plantarum 17-51 x 106 CFU/mL0% (0/3)0% (0/3)
Legionella longbeachae1 x 106 CFU/mL0% (0/3)0% (0/3)
Cross ReactantConcentrationBordetella pertussis(IS481)% Detection(# Detected/# Tested)Bordetella parapertussis(IS1001)% Detection(# Detected/# Tested)
Legionella pneumophila(Philadelphia)1 x 106 CFU/mL0% (0/3)0% (0/3)
Listeria monocytogenes1 x 106 CFU/mL0% (0/3)0% (0/3)
Measles1 x 105 TCID50/mL0% (0/3)0% (0/3)
Metapneumovirus-91 x 105 TCID50/mL0% (0/3)0% (0/3)
Moraxella catarrhalis Ne 111 x 106 CFU/mL0% (0/3)0% (0/3)
Morganella morganii1 x 106 CFU/mL0% (0/3)0% (0/3)
Mumps1 x 105 TCID50/mL0% (0/3)0% (0/3)
Mycobacterium avium1 x 106 CFU/mL0% (0/3)0% (0/3)
Mycobacterium tuberculosis(genomic DNA)1 x 106 genomecopies/mL0% (0/3)0% (0/3)
Mycoplasma hominis1 x 106 CFU/mL0% (0/3)0% (0/3)
Mycoplasma pneumoniae StrainM1291 x 106 CCU/mL0% (0/3)0% (0/3)
Neisseria elongata1 x 106 CFU/mL0% (0/3)0% (0/3)
Neisseria gonorrhoeae1 x 106 CFU/mL0% (0/3)0% (0/3)
Neisseria meningitidis1 x 106 CFU/mL0% (0/3)0% (0/3)
Neisseria mucosa1 x 106 CFU/mL0% (0/3)0% (0/3)
Parainfluenza 11 x 105 TCID50/mL0% (0/3)0% (0/3)
Parainfluenza 21 x 105 TCID50/mL0% (0/3)0% (0/3)
Parainfluenza 3*1 x 104 TCID50/mL0% (0/3)0% (0/3)
Parainfluenza 4*1 x 104 TCID50/mL0% (0/3)0% (0/3)
Parvimonas micra1 x 106 CFU/mL0% (0/3)0% (0/3)
Peptostreptococcus anaerobius1 x 106 CFU/mL0% (0/3)0% (0/3)
Proteus mirabilis Z0501 x 106 CFU/mL0% (0/3)0% (0/3)
Proteus vulgaris1 x 106 CFU/mL0% (0/3)0% (0/3)
Pseudomonas aeruginosa1 x 106 CFU/mL0% (0/3)0% (0/3)
Pseudomonas fluorescens1 x 106 CFU/mL0% (0/3)0% (0/3)
Rhinovirus 1A*1 x 104 TCID50/mL0% (0/3)0% (0/3)
RSV A1 x 105 TCID50/mL0% (0/3)0% (0/3)
RSV B WV/14617/851 x 105 TCID50/mL0% (0/3)0% (0/3)
Serratia liquefaciens1 x 106 CFU/mL0% (0/3)0% (0/3)
Serratia marcescens1 x 106 CFU/mL0% (0/3)0% (0/3)
Staphylococcus aureus (MRSA)1 x 106 CFU/mL0% (0/3)0% (0/3)
Staphylococcus epidermidis(MRSE)1 x 106 CFU/mL0% (0/3)0% (0/3)
Stenotrophomonas maltophilia1 x 106 CFU/mL0% (0/3)0% (0/3)
Streptococcus anginosus1 x 106 CFU/mL0% (0/3)0% (0/3)
Streptococcus canis1 x 106 CFU/mL0% (0/3)0% (0/3)
Cross ReactantConcentrationBordetella pertussis(IS481)% Detection(# Detected/# Tested)Bordetella parapertussis(IS1001)% Detection(# Detected/# Tested)
Streptococcus dysgalactiae1 x 106 CFU/mL0% (0/3)0% (0/3)
Streptococcus intermedius1 x 106 CFU/mL0% (0/3)0% (0/3)
Streptococcus mitis1 x 106 CFU/mL0% (0/3)0% (0/3)
Streptococcus mutans1 x 106 CFU/mL0% (0/3)0% (0/3)
Streptococcus pneumoniae1 x 106 CFU/mL0% (0/3)0% (0/3)
Streptococcus pyogenes M11 x 106 CFU/mL0% (0/3)0% (0/3)
Streptococcus salivarius1 x 106 CFU/mL0% (0/3)0% (0/3)
Ureaplasma urealyticum1 x 106 CCU/mL0% (0/3)0% (0/3)
Varicella Zoster Virus*1 x 104 TCID50/mL0% (0/3)0% (0/3)

{12}------------------------------------------------

Image /page/12/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, the word "DiaSorin" is written in a dark blue, sans-serif font, with the word "Molecular" underneath in a lighter green color.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 10 of 17

{13}------------------------------------------------

Image /page/13/Picture/0 description: The image contains the logo for DiaSorin Molecular. The logo features a stylized DNA strand in shades of green and blue on the left. To the right of the DNA strand 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™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 11 of 17

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

Image /page/14/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a stylized DNA double helix on the left, rendered in shades of green and blue. To the right of the helix, the word "DiaSorin" is written in a dark blue, sans-serif font. Below "DiaSorin", the word "Molecular" is written in a lighter green, sans-serif font.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 12 of 17

  • The testing concentration of these viruses is lower due to the lack of a high titer stock.

INTERFERENCE

The performance of Simplexa™ Bordetella Direct was evaluated with potentially interfering substances that may be present in the nasopharynx. The potentially interfering substances were evaluated in a contrived sample that contained Bordetella pertussis and Bordetella parapertussis at approximately 2-4 X LoD. There was no evidence of interference caused by the substances at the concentrations listed in Table 9 (Bordetella pertussis) and Table 10 (Bordetella parapertussis).

Table 9. Simplexa™ Bordetella Direct Interference - Bordetella pertussis
--------------------------------------------------------------------------------
Potentially Interfering SubstanceActive IngredientInterferentConcentrationB. pertussis (IS481)% Detection
Albuterol sulfateAlbuterol sulfate10 mg/mL100% (3/3)
Ampicillin powderAmpicillin10 mg/mL100% (3/3)
Azithromycin powderAzithromycin10 mg/mL100% (3/3)
Beclomethasone dipropionateBeclomethasonedipropionate10 mg/mL100% (3/3)
BloodNA10% v/v100% (3/3)
Chloraseptic sore throat sprayPhenol10% v/v100% (3/3)
CiprofloxacinCiprofloxacin1.25 mg/mL100% (3/3)
ErythromycinErythromycin10 mg/mL100% (3/3)
Flonase Nasal SprayFluticasone propionate-corticosteroid10% v/v100% (3/3)
MucinMucin10 mg/mL100% (3/3)
MupirocinMupirocin10 mg/mL100% (3/3)
RifampicinRifampicin2.5 mg/mL100% (3/3)
Robitussin DMRobitussin DM10% v/v100% (3/3)
Saline Nasal spray-SodiumchlorideSodium chloride10% v/v100% (3/3)

{15}------------------------------------------------

Image /page/15/Picture/0 description: The image contains the word "orin" in a dark blue font. The letters are bold and slightly spaced apart. There is a dot above the "i" in "orin."

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 13 of 17

Potentially Interfering SubstanceActive IngredientInterferentConcentrationB. pertussis (IS481)% Detection
Sudafed PEPhenylephrine10 mg/mL100% (3/3)
Zicam 12 hrs sprayOxymetazoline HCl10% v/v100% (3/3)

Table 10. Simplexa™ Bordetella Direct Interference - Bordetella parapertussis

Potentially Interfering SubstanceActive IngredientInterferentConcentrationB. parapertussis(IS1001)% Detection
Albuterol sulfateAlbuterol sulfate10 mg/mL100% (3/3)
Ampicillin powderAmpicillin10 mg/mL100% (3/3)
Azithromycin powderAzithromycin10 mg/mL100% (3/3)
Beclomethasone dipropionateBeclomethasonedipropionate10 mg/mL100% (3/3)
BloodNA10% v/v100% (3/3)
Chloraseptic sore throat sprayPhenol10% v/v100% (3/3)
CiprofloxacinCiprofloxacin1.25 mg/mL100% (3/3)
ErythromycinErythromycin10 mg/mL100% (3/3)
Flonase Nasal SprayFluticasone propionate-corticosteroid10% v/v100% (3/3)
MucinMucin10 mg/mL100% (3/3)
MupirocinMupirocin10 mg/mL100% (3/3)
RifampicinRifampicin5 mg/mL100% (8/8)*
Robitussin DMRobitussin DM10% v/v100% (3/3)
Saline Nasal spray-SodiumchlorideSodium chloride10% v/v100% (3/3)
Sudafed PEPhenylephrine10 mg/mL100% (3/3)
Zicam 12 hrs sprayOxymetazoline HCl10% v/v100% (3/3)
  • The initial three (3) replicates of Rifampicin at 5 mg/mL were valid but not detected for Bordetella parapertussus. An additional five (5) replicates were performed with no interference for Bordetella parapertussis at 5 mg/mL.

COMPETITIVE INTERFERENCE

The Simplexa™ Bordetella Direct assay was evaluated for competitive interference by testing whether the presence of a clinically relevant high concentration of either Bordetella pertussis or Bordetella parapertussis could affect the detection of the other Bordetella species when present at a low level. A low positive sample was contrived for each target by spiking Bordetella pertussis or Bordetella parapertussis separately (at approximately 2 X LoD) into nasopharyngeal swab matrix in Universal Transport Media (UTM) and a baseline Ct was determined for each sample. High concentrations of the other Bordetella strain were then spiked into the low level sample and tested. No competitive interference was observed. The results showed that a low level of Bordetella pertussis was detected in the presence of a high level of Bordetella parapertussis; similarly, a low level of Bordetella parapertussis was detected in the presence of a high level of Bordetella pertussis. The results are shown in Table 11.

{16}------------------------------------------------

Image /page/16/Picture/1 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, the word "DiaSorin" is written in a dark blue, sans-serif font. Below "DiaSorin", the word "Molecular" is written in a lighter green, sans-serif font.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 14 of 17

Baseline(Low Concentration)Competitive Interferent(High Concentration)Bordetella pertussis(IS481)(#Detected/#Total)Bordetella parapertussis(IS1001)(#Detected/#Total)
Species2 X LoDCFU/mLSpeciesCFU/mL
Bordetella pertussis29.4 CFU/mLBordetella parapertussis2x107 CFU/mL3/33/3
Bordetella parapertussis694.6 CFU/mLBordetella pertussis1X107 CFU/mL3/33/3

Table 11. Simplexa™ Bordetella Direct Competitive Interference

INHIBITION BY OTHER MICROORGANISMS

Simplexa™ Bordetella Direct was tested for the ability to identify Bordetella when potentially inhibitory organisms were present. The panel of ninety-seven (97) potentially inhibitory organisms was individually spiked into a pool with a low concentration (approximately 2X LoD) of Bordetella pertussis and Bordetella parapertussis. The organisms were spiked into the matrix at the concentrations indicated in Table 12. Bordetella pertussis and Bordetella parapertussis were tested separately (data not shown) and as a dual Below are the results of the dual positive Bordetella pertussis and Bordetella positive sample. parapertussis sample in Table 12. No inhibitory effects were observed for Simplexa™ Bordetella Direct at the concentrations tested.

Table 12. Summary of Results for the Microbial Inhibition Study
Bordetella pertussis and Bordetella parapertussis at 2 X LoD
OrganismConcentrationBordetellapertussis (IS481)% Detection(# Detected/# Tested)Bordetellaparapertussis (IS1001)% Detection(# Detected/# Tested)
BaselineN/A100% (45/45)100% (45/45)
Acinetobacter baumannii1 x 106 CFU/mL100% (3/3)100% (3/3)
Acinetobacter Iwoffi1 x 106 CFU/mL100% (3/3)100% (3/3)
Adenovirus 11 x 105 TCID50/mL100% (3/3)100% (3/3)
Adenovirus 311 x 105 TCID50/mL100% (3/3)100% (3/3)
Arcanobacterium haemolyticum1 x 106 CFU/mL100% (3/3)100% (3/3)
Bacillus cereus1 x 106 CFU/mL100% (3/3)100% (3/3)
Bacteroides fragilis1 x 106 CFU/mL100% (3/3)100% (3/3)
Bordetella avium1 x 106 CFU/mL100% (3/3)100% (3/3)
Bordetella bronchiseptica1 x 106 CFU/mL100% (3/3)100% (3/3)
Bordetella hinzi1 x 106 CFU/mL100% (3/3)100% (3/3)
Bordetella holmseii F0611 x 106 CFU/mL100% (3/3)100% (3/3)
Bordetella petri1 x 106 CFU/mL100% (3/3)100% (3/3)
Bordetella trematum1 x 106 CFU/mL100% (3/3)100% (3/3)
Burkholderia cenocepacia1 x 106 CFU/mL100% (3/3)100% (3/3)
OrganismConcentrationBordetellapertussis (IS481)% Detection(# Detected/# Tested)Bordetellaparapertussis (IS1001)% Detection(# Detected/# Tested)
Burkholderia cepacia1 x 10° CFU/mL100% (3/3)100% (3/3)
Burkholderia multivorans1 x 10° CFU/mL100% (3/3)100% (3/3)
Burkholderia thailandensis1 x 10° CFU/mL100% (3/3)100% (3/3)
Candida albicans1 x 10° CFU/mL100% (3/3)100% (3/3)
Candida glabrata1 x 106 CFU/mL100% (3/3)100% (3/3)
Chlamydia pneumoniae1 x 10° IFU/mL100% (3/3)100% (3/3)
Chlamydia trachmomatis1 x 10° IFU/mL100% (3/3)100% (3/3)
Citrobacter freundii1 x 10° CFU/mL100% (3/3)100% (3/3)
Clostridium difficile1 x 10° CFU/mL100% (3/3)100% (3/3)
Coronavirus 229E1 x 105 TCID50/mL100% (3/3)100% (3/3)
Coronavirus NL63*1 x 104 TCID50/mL100% (3/3)100% (3/3)
Coronavirus OC431 x 10° TCID50/mL100% (3/3)100% (3/3)
Corynebacterium diptheriae1 x 10° CFU/mL100% (3/3)100% (3/3)
Coxsackievirus A161 x 105 TCID50/mL100% (3/3)100% (3/3)
Coxsackievirus B41 x 105 TCID50/mL100% (3/3)100% (3/3)
Cytomegalovirus1 x 105 TCID50/mL100% (3/3)100% (3/3)
Echovirus 61 x 105 TCID50/mL100% (3/3)100% (3/3)
Echovirus 71 x 105 TCID50/mL100% (3/3)100% (3/3)
Echovirus 91 x 10° TCID50/mL100% (3/3)100% (3/3)
Echovirus 111 x 10 TCID50/mL100% (3/3)100% (3/3)
Enterobacter aerogenes Z0521 x 10° CFU/mL100% (3/3)100% (3/3)
Enterobacter cloacae1 x 10° CFU/mL100% (3/3)100% (3/3)
Enterococcus faecalis vanB1 x 106 CFU/mL100% (3/3)100% (3/3)
Enterovirus 701 x 105 TCID50/mL100% (3/3)100% (3/3)
Enterovirus 711 x 10 TCID50/mL100% (3/3)100% (3/3)
Epstein-Barr Virus1 x 10° copies/mL100% (3/3)100% (3/3)
Escherichia coli1 x 10° CFU/mL100% (3/3)100% (3/3)
Fusobacterium necrophorum1 x 10° CFU/mL100% (3/3)100% (3/3)
Haemophilus influenzae1 x 10° CFU/mL100% (3/3)100% (3/3)
Haemophilus parainfluenzae1 x 10° CFU/mL100% (3/3)100% (3/3)
HSV-1 (MacIntyre)1 x 105 TCID50/mL100% (3/3)100% (3/3)
HSV-2 (G)1 x 10 TCID50/mL100% (3/3)100% (3/3)
Influenza A/Swine/lowa/15/30H1N1*1 x 104 TCID50/mL100% (3/3)100% (3/3)
OrganismConcentrationBordetellapertussis (IS481)% Detection(# Detected/# Tested)Bordetellaparapertussis (IS1001)% Detection(# Detected/# Tested)
Influenza B/Malaysia/2506/04*1 x 104 TCID50/mL100% (3/3)100% (3/3)
Klebsiella oxytoca1 x 106 CFU/mL100% (3/3)100% (3/3)
Klebsiella pneumoniae1 x 106 CFU/mL100% (3/3)100% (3/3)
Lactobacillus acidophilus1 x 106 CFU/mL100% (3/3)100% (3/3)
Lactobacillus plantarum 17-51 x 106 CFU/mL100% (3/3)100% (3/3)
Legionella longbeachae1 x 106 CFU/mL100% (3/3)100% (3/3)
Legionella pneumophila(Philadelphia)1 x 106 CFU/mL100% (3/3)100% (3/3)
Listeria monocytogenes1 x 106 CFU/mL100% (3/3)100% (3/3)
Measles1 x 105 TCID50/mL100% (3/3)100% (3/3)
Metapneumovirus-91 x 105 TCID50/mL100% (3/3)100% (3/3)
Moraxella catarrhalis Ne 111 x 106 CFU/mL100% (3/3)100% (3/3)
Morganella morganii1 x 106 CFU/mL100% (3/3)100% (3/3)
Mumps1 x 105 TCID50/mL100% (3/3)100% (3/3)
Mycobacterium avium1 x 106 CFU/mL100% (3/3)100% (3/3)
Mycobacterium tuberculosis(genomic DNA)1 x 106 genomecopies/mL100% (3/3)100% (3/3)
Mycoplasma hominis1 x 106 CFU/mL100% (3/3)100% (3/3)
Mycoplasma pneumoniae StrainM1291 x 106 CCU/mL100% (3/3)100% (3/3)
Neisseria elongata1 x 106 CFU/mL100% (3/3)100% (3/3)
Neisseria gonorrhoeae1 x 106 CFU/mL100% (3/3)100% (3/3)
Neisseria meningitidis1 x 106 CFU/mL100% (3/3)100% (3/3)
Neisseria mucosa1 x 106 CFU/mL100% (3/3)100% (3/3)
Parainfluenza 11 x 105 TCID50/mL100% (3/3)100% (3/3)
Parainfluenza 21 x 105 TCID50/mL100% (3/3)100% (3/3)
Parainfluenza 3*1 x 104 TCID50/mL100% (3/3)100% (3/3)
Parainfluenza 4*1 x 104 TCID50/mL100% (3/3)100% (3/3)
Parvimonas micra1 x 106 CFU/mL100% (3/3)100% (3/3)
Peptostreptococcus anaerobius1 x 106 CFU/mL100% (3/3)100% (3/3)
Proteus mirabilis Z0501 x 106 CFU/mL100% (3/3)100% (3/3)
Proteus vulgaris1 x 106 CFU/mL100% (3/3)100% (3/3)
Pseudomonas aeruginosa1 x 106 CFU/mL100% (3/3)100% (3/3)
Pseudomonas fluorescens1 x 106 CFU/mL100% (3/3)100% (3/3)
Rhinovirus 1A*1 x 104 TCID50/mL100% (3/3)100% (3/3)
OrganismConcentrationBordetellapertussis (IS481)% Detection(# Detected/# Tested)Bordetellaparapertussis (IS1001)% Detection(# Detected/# Tested)
RSV A1 x 105 TCID50/mL100% (3/3)100% (3/3)
RSV B WV/14617/851 x 108 TCID50/mL100% (3/3)100% (3/3)
Serratia liquefaciens1 x 10° CFU/mL100% (3/3)100% (3/3)
Serratia marcescens1 x 10° CFU/mL100% (3/3)100% (3/3)
Staphylococcus aureus (MRSA)1 x 10° CFU/mL100% (3/3)100% (3/3)
Staphylococcus epidermidis(MRSE)1 x 10° CFU/mL100% (3/3)100% (3/3)
Stenotrophomonas maltophilia1 x 10° CFU/mL100% (3/3)100% (3/3)
Streptococcus anginosus1 x 10° CFU/mL100% (3/3)100% (3/3)
Streptococcus canis1 x 10° CFU/mL100% (3/3)100% (3/3)
Streptococcus dysgalactiae1 x 10° CFU/mL100% (3/3)100% (3/3)
Streptococcus intermedius1 x 10° CFU/mL100% (3/3)100% (3/3)
Streptococcus mitis1 x 10° CFU/mL100% (3/3)100% (3/3)
Streptococcus mutans1 x 10° CFU/mL100% (3/3)100% (3/3)
Streptococcus pneumoniae1 x 10° CFU/mL100% (3/3)100% (3/3)
Streptococcus pyogenes M11 x 10° CFU/mL100% (3/3)100% (3/3)
Streptococcus salivarius1 x 10° CFU/mL100% (3/3)100% (3/3)
Ureaplasma urealyticum1 x 10° CCU/mL100% (3/3)100% (3/3)
Varicella Zoster Virus*1 x 104 TCID50/mL100% (3/3)100% (3/3)

{17}------------------------------------------------

Image /page/17/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a green and blue DNA helix graphic on the left, with the text "DiaSorin" in dark blue on the top right and "Molecular" in green below it. The text is in a serif font and is aligned to the right of the DNA helix graphic.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 15 of 17

{18}------------------------------------------------

Image /page/18/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 a dark blue serif font, with the word "Molecular" underneath in a lighter green sans-serif font.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018
Page 16 of 17

{19}------------------------------------------------

Image /page/19/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA strand in shades of green and blue on the left. To the right of the DNA strand are the words "DiaSorin" in dark blue on the top line and "Molecular" in green on the bottom line.

510(k) Summary

Simplexa™ Bordetella Direct Catalog No. MOL2750 Simplexa™ Bordetella Positive Control Pack Catalog No. MOL2760 August 7, 2018 Page 17 of 17

  • The testing concentration of these viruses is lower due to the lack of a high titer stock.

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.

§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.

(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.