Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on real-time PCR technology and standard laboratory methods for detection and confirmation, with no mention of AI or ML.

Therapeutic

No
The device is an in vitro diagnostic (IVD) assay intended for the qualitative detection of Group B Streptococcus nucleic acid, used as an aid in determining colonization status. It is explicitly stated that the assay results "are not intended to diagnose or monitor treatment of a GBS infection," which means it does not treat or prevent disease, thus it is not a therapeutic device.

Diagnostic

Yes

The device qualitatively detects Group B Streptococcus (GBS) nucleic acid, which helps determine the colonization status of antepartum women. This falls under the definition of a diagnostic device as it aids in identifying a biological condition.

SaMD (Software as a Medical Device)

No

The device description explicitly states that the system consists of the Simplexa™ GBS Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc (DAD) and associated accessories. This includes hardware components like the LIAISON® MDX instrument and the DAD, making it a system that includes both hardware and software, not a software-only device.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the assay is "intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of Group B Streptococcus (GBS) nucleic acid". The term "in vitro" is a key indicator of an IVD, meaning it's used outside of a living organism.
Device Description: The description details a "real-time PCR system" that analyzes "bacterial DNA from vaginal swabs enriched in Lim Broth". This process involves analyzing biological samples outside the body to provide diagnostic information.
Performance Studies: The document describes clinical performance studies comparing the assay to a "GBS culture reference method", which is typical for validating an IVD.
Key Metrics: The inclusion of metrics like Sensitivity and Specificity are standard for evaluating the performance of diagnostic tests.

Therefore, based on the provided information, the Simplexa™ GBS Direct assay clearly fits the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Simplexa™ GBS Direct
The DiaSorin Molecular Simplexa™ GBS Direct assay is a real-time polymerase chain reaction (PCR) assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of Group B Streptococcus (GBS) nucleic acid from 18 to 24 hour Lim broth enrichments of vaginal/rectal specimen swabs obtained from antepartum women. Assay results can be used as an aid in determining the colonization status of antepartum women, but are not intended to diagnose or monitor treatment of a GBS infection.

The Simplexa™ GBS Direct assay does not provide susceptibility results. Culture isolates are needed to perform susceptibility testing as recommended for penicillin-allergic women.

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

Product codes (comma separated list FDA assigned to the subject device)

NJR, OOI

Device Description

The Simplexa™ GBS Direct assay system is a real-time PCR system that enables the direct amplification and qualitative detection of Group B Strep bacterial DNA from vaginal swabs enriched in Lim Broth for eighteen to twenty-four (18 to 24) hours that have not undergone a nucleic acid extraction. The system consists of the Simplexa™ GBS Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc (DAD) and associated accessories.

In the Simplexa™ GBS Direct assay, primers and fluorescent probes are used together to amplify Group B Streptococcus bacterial DNA and the Internal Control (DNA IC). The assay targets a conserved region of the cfb gene to identify Group B Streptococus in the specimen. The DNA IC is used to detect PCR failure and/or inhibition.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

vaginal/rectal specimen swabs

Indicated Patient Age Range

antecedant women

Intended User / Care Setting

Not Found

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

Not Found

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

The performance of the Simplexa™ GBS Direct assay was evaluated in a prospective study that was conducted with residual de-identified Lim broth enrichment cultures of vaginal/rectal swabs obtained from pregnant women at thirty-five to thirty-seven (35-37) weeks of gestation. The specimens were collected and enriched according to established CDC guidelines. Lim Broth cultures were incubated for eighteen to twenty-four (18-24) hours at 35-37 °C. Four hundred and thirty-two (432) samples that met the prescribed inclusion criteria were used to evaluate the performance of the Simplexa™ GBS Direct assay in comparison to those obtained with a GBS culture reference method. All samples were tested fresh upon completion of the enrichment process. Aliquots were made from each freshly enriched Lim Broth sample; the first aliquot was held at 2-8 °C and tested on the Simplexa™ GBS Direct at the collection site, and the second aliquot was kept at 2-8 °C and sent on cold packs to a central laboratory for GBS culture.

For the GBS culture reference method, enriched Lim Broth samples were sub-cultured to selective and non-selective blood agar for twenty-four (24) hours. If GBS was not observed, the plates were reincubated for an additional twenty-four (24) hours and deemed negative if GBS was not identified. Colonies with an appearance that was suggestive of GBS were further processed to confirm the presence of GBS using general laboratory methods, which included: gram stain, catalase testing, and latex agglutination.

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

Clinical Performance: A prospective study with 432 samples of residual de-identified Lim broth enrichment cultures of vaginal/rectal swabs. The Simplexa™ GBS Direct assay results were compared to a GBS culture reference method. The invalid rate was 0.0% (0/432).
Reproducibility: Evaluated across 3 investigative sites for inter-day and inter/intra-assay reproducibility. Each laboratory tested Simplexa™ GBS Direct Positive Control, No Template Control (Lim Broth), and 4 contrived samples (low positive and medium positive for two GBS strains: BAA-22 and BAA-1138) in negative matrix. Assays were performed in triplicate on 5 different days. Two operators per site, total of 6 instruments. Total N = 540 for both GBS and Internal Control (Q670).
Analytical Sensitivity/Limit of Detection (LoD): Determined using quantified stocks of two GBS strains (BAA-22 and BAA-1138) serially diluted into negative 18-24 hour Lim broth enrichments. LoD was the lowest concentration detected positive >= 95% of the time. LoD for ATCC BAA-22 (serotype III) was 80,000 CFU/mL. LoD for ATCC BAA-1138 (serotype Ia) was 30,000 CFU/mL.
Analytical Reactivity: Evaluated against 18 additional GBS strains spiked at 2 x LoD into 18-24 hour Lim broth enrichments. All 18 strains were detected (100% detection for each, 3/3 tests).
Cross Reactivity (Analytical Specificity): Evaluated by testing 74 potential cross-reacting organisms spiked into GBS negative enriched Lim broth in triplicate (4 organisms tested in silico). No cross-reactivity was observed.
Interference: Evaluated 26 potentially interfering endogenous and exogenous substances spiked into baseline samples (GBS BAA-1138 or BAA-22 at 2 x LoD) in 18-24 hour Lim broth enrichments. No interference was observed at the indicated concentrations.
Inhibition by other microorganisms: Evaluated the ability to identify GBS when 74 potentially inhibitory organisms were present at high concentrations (spiked individually into a pool with GBS BAA-22 or BAA-1138 at 2 x LoD). No false negative results with GBS BAA-22. With GBS BAA-1138, false negatives were observed in presence of Bacillus cereus, Candida parapsilosis, Clostridium perfringens, Peptostreptococcus anaerobius, Staphylococcus saprophyticus, Streptococcus pyogenes, and Ureaplasma urealyticum. In silico analysis showed no potential for cross-reaction or interference from Norovirus genogroups I and II or Trichomonas vaginalis primers/probe.
Carry-Over Contamination: Assessed by testing alternating high positive and negative samples on each disc. No evidence observed.

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

Clinical Agreement:
Sensitivity: 97.0% (97/100) (95% CI: 91.5% to 99.0%)
Specificity: 96.1% (319/332) (95% CI: 93.4% to 97.7%)

Reproducibility (GBS (FAM) agreement with expected results):
BAA-22 - LP: 100.0% (90/90) (95% CI: 95.9% to 100.0%)
BAA-22 - MP: 100.0% (90/90) (95% CI: 95.9% to 100.0%)
BAA-1138 - LP: 97.8% (88/90) (95% CI: 92.3% to 99.4%)
BAA-1138 - MP: 100.0% (90/90) (95% CI: 95.9% to 100.0%)
PC: 100.0% (90/90) (95% CI: 95.9% to 100.0%)
NTC: 100.0% (90/90) (95% CI: 95.9% to 100.0%)
Total Agreement: 99.6% (538/540) (95% CI: 98.7% to 99.9%)

Reproducibility (Internal Control (Q670) agreement with expected results):
All individual samples (BAA-22 LP, BAA-22 MP, BAA-1138 LP, BAA-1138 MP, PC, NTC) showed 100.0% agreement (90/90) with 95% CI: 95.9% to 100.0%.
Total Agreement: 100.0% (540/540) (95% CI: 99.3% to 100.0%)

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

GenePOC GBS LB Assay K170557

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

Regulation Number and Section

§ 866.3740

Streptococcus spp. serological reagents.(a)
Identification. Streptococcus spp. serological reagents are devices that consist of antigens and antisera (excluding streptococcal exoenzyme reagents made from enzymes secreted by streptococci) used in serological tests to identifyStreptococcus spp. from cultured isolates derived from clinical specimens. The identification aids in the diagnosis of diseases caused by bacteria belonging to the genusStreptococcus and provides epidemiological information on these diseases. Pathogenic streptococci are associated with infections, such as sore throat, impetigo (an infection characterized by small pustules on the skin), urinary tract infections, rheumatic fever, and kidney disease.(b)
Classification. Class I (general controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to § 866.9.

Summary

0

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

November 7, 2018

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

Re: K182467

Trade/Device Name: Simplexa GBS Direct, Simplexa GBS Positive Control Pack Regulation Number: 21 CFR 866.3740 Regulation Name: Streptococcus spp. Serological Reagents Regulatory Class: Class I Product Code: NJR, OOI Dated: September 5, 2018 Received: September 10, 2018

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/cdrl/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 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR

1

for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (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 http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

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/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Steven R. Gitterman -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) K182467

Device Name Simplexa™ GBS Direct Simplexa™ GBS Positive Control Pack

Indications for Use (Describe)

Simplexa™ GBS Direct

The DiaSorin Molecular Simplexa™ GBS Direct assay is a real-time polymerase chain reaction (PCR) assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of Group B Streptococcus (GBS) nucleic acid from 18 to 24 hour Lim broth enrichments of vaginal/rectal specimen swabs obtained from antepartum women. Assay results can be used as an aid in determining the colonization status of antepartum women, but are not intended to diagnose or monitor treatment of a GBS infection.

The Simplexa™ GBS Direct assay does not provide susceptibility results. Culture isolates are needed to perform susceptibility testing as recommended for penicillin-allergic women.

Simplexa™ GBS Positive Control Pack

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

Type of Use (Select one or both, as applicable)
Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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

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Image /page/3/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in dark blue, with "Molecular" underneath in green. The logo is clean and modern, representing the company's focus on molecular diagnostics.

510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 1 of 19

| Applicant | DiaSorin Molecular LLC.
11331 Valley View Street
Cypress, California 90630
USA |
|--------------------------------|------------------------------------------------------------------------------------------------------------------------------|
| Establishment Registration No. | 2023365 |
| Contact Person | Sharon Young
Principal Regulatory Affairs Specialist
tel 562.240.6680
fax 562.240.6529
Sharon.Young@DiaSorin.com |
| Summary Date | October 25 2018 |
| Proprietary Name | Simplexa™ GBS Direct and Simplexa™ GBS Positive Control Pack |
| Generic Name | Nucleic Acid Amplifications System, Group B Streptococcus, Direct
Specimen Testing |
| Classification | Class I |
| Predicate Devices | GenePOC GBS LB Assay K170557 |

Intended Use

Simplexa™ GBS Direct

The DiaSorin Molecular Simplexa™ GBS Direct assay is a real-time polymerase chain reaction (PCR) assay intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of Group B Streptococcus (GBS) nucleic acid from 18 to 24 hour Lim broth enrichments of vaginal/rectal specimen swabs obtained from antepartum women. Assay results can aid in determining the colonization status of antepartum women, but are not intended to diagnose or monitor treatment of a GBS infection.

The Simplexa™ GBS Direct assay does not provide susceptibility results. Culture isolates are needed to perform susceptibility testing as recommended for penicillin-allergic women.

Simplexa™ GBS Positive Control Pack

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

Device Description

The Simplexa™ GBS Direct assay system is a real-time PCR system that enables the direct amplification and qualitative detection of Group B Strep bacterial DNA from vaginal swabs enriched in Lim Broth for eighteen to twenty-four (18 to 24) hours that have not undergone a nucleic acid extraction. The system consists of the Simplexa™ GBS Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc (DAD) and associated accessories.

In the Simplexa™ GBS Direct assay, primers and fluorescent probes are used together to amplify Group B Streptococcus bacterial DNA and the Internal Control (DNA IC). The assay targets a conserved region of the cfb gene to identify Group B Streptococus in the specimen. The DNA IC is used to detect PCR failure and/or inhibition.

| Component Name | REF | EC SYMBOL
ON LABEL | Abbreviated
Name | Cap
Color | Number of
Vials | Reactions
per
Vial/Kit | Volume
per Vial |
|-----------------------------------------|---------|-----------------------|---------------------|--------------|--------------------|------------------------------|--------------------|
| Simplexa™ GBS
Direct
Reaction Mix | MOL3551 | REAG
C | RM | Orange | 24 | 1/24 | 50 μL |

Kit Description

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Image /page/4/Picture/0 description: The image contains the logo for DiaSorin Molecular. The logo features a stylized DNA double helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in a dark blue, sans-serif font, stacked above the word "Molecular" in a lighter green, sans-serif font.

510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 2 of 19

Component Description

Kit Component	Contents
Simplexa™ GBS
Direct Reaction
Mix (RM)	DNA polymerase, buffers, dNTPs, Internal Control DNA Template, dye-labeled fluorescent
primers and probes specific for detection of Group B Strep and for the DNA Internal Control
	Target	Probe
Fluorophore
(Dye)	Excitation	Emission	Targeted Gene
	GBS	CFR610	590	610	cfb
	Internal
Control
DNA (IC)	Q670	644	670	NA
Simplexa™ GBS
Direct Barcode
Card	Assay specific parameters, lot number, expiration date.

MATERIALS SUPPLIED SEPARATELY

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

Predicate Device Information

Similarities
Comparison to
Predicate Device	Predicate Device:
GenePOC GBS LB Assay K170557	Candidate Device:
Simplexa™ GBS Direct and
Simplexa™ GBS Positive Control Pack
Product Code	NJR and OOI
21 CFR 866.3740 - Streptococcal spp.
serological reagents
21 CFR 862.2570 - Instrumentation for
clinical multiplex test systems	Same
Organism
Detected	Group B Streptococcus	Same
Measurand	Target DNA sequence in cfb gene of
Streptococcus agalactiae (Group B
Streptococcus, GBS)	Conserved region of the cfb gene of
Streptococcus agalactiae (Group B
Streptococcus, GBS)
Intended Use	The GenePOC GBS LB assay
performed on the revogene instrument
is a qualitative in vitro diagnostic test
designed to detect Group B
Streptococcus (GBS) DNA from 18-24
hour LIM broth enrichments of
vaginal/rectal specimen swabs	The DiaSorin Molecular Simplexa™ GBS
Direct assay is a real-time polymerase
chain reaction (PCR) assay intended for
use on the LIAISON® MDX instrument
for the in vitro qualitative detection of
Group B Streptococcus (GBS) nucleic
acid from 18 to 24 hour Lim broth

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Image /page/5/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a stylized DNA double helix in shades of green on the left. To the right of the DNA helix are the words "DiaSorin" in dark blue on the top line and "Molecular" in light green on the bottom line.

510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 3 of 19

Similarities
Comparison to
Predicate Device	Predicate Device:
GenePOC GBS LB Assay K170557	Candidate Device:
Simplexa™ GBS Direct and
Simplexa™ GBS Positive Control Pack
	obtained from pregnant women. The
GenePOC GBS LB assay utilizes
automated sample processing and
real-time polymerase chain reaction
(PCR) to detect a cfb gene sequence
specific to the Streptococcus
agalactiae genome. The GenePOC
GBS LB assay is indicated for the
identification of antepartum GBS
colonization and does not provide
susceptibility results. It is not intended
to diagnose or monitor treatment of
GBS infection. Culture isolates are
needed for performing susceptibility
testing as recommended for penicillin-
allergic women.	enrichments of vaginal/rectal specimen
swabs obtained from antepartum
women. Assay results can be used as an
aid in determining the colonization status
of antepartum women, but are not
intended to diagnose or monitor
treatment of a GBS infection.

The Simplexa™ GBS Direct assay does
not provide susceptibility results. Culture
isolates are needed to perform
susceptibility testing as recommended for
penicillin-allergic women.

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

This control is not intended for use with
other assays or systems. |
| Automated
System (Sample
to Answer) | Yes | Yes |

Differences
Comparison to
Predicate
Device	GenePOC GBS LB Assay K170557	Simplexa™ GBS Direct and
Simplexa™ GBS Positive Control
Pack
Instrumentation	Revogene instrument	LIAISON® MDX
Single Use	Yes Cartridge (PIE) can be used once.	No. Consumable disc can be used until
all eight (8) wedges have been used.

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Image /page/6/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 on the top line and "Molecular" in green on the bottom line. The text is aligned to the right of the DNA helix.

510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 4 of 19

CLINICAL PERFORMANCE

The performance of the Simplexa™ GBS Direct assay was evaluated in a prospective study that was conducted with residual de-identified Lim broth enrichment cultures of vaginal/rectal swabs obtained from pregnant women at thirty-five to thirty-seven (35-37) weeks of gestation. The specimens were collected and enriched according to established CDC guidelines. Lim Broth cultures were incubated for eighteen to twenty-four (18-24) hours at 35-37 °C. Four hundred and thirty-two (432) samples that met the prescribed inclusion criteria were used to evaluate the performance of the Simplexa™ GBS Direct assay in comparison to those obtained with a GBS culture reference method. All samples were tested fresh upon completion of the enrichment process. Aliquots were made from each freshly enriched Lim Broth sample; the first aliquot was held at 2-8 °C and tested on the Simplexa™ GBS Direct at the collection site, and the second aliquot was kept at 2-8 °C and sent on cold packs to a central laboratory for GBS culture.

For the GBS culture reference method, enriched Lim Broth samples were sub-cultured to selective and non-selective blood agar for twenty-four (24) hours. If GBS was not observed, the plates were reincubated for an additional twenty-four (24) hours and deemed negative if GBS was not identified. Colonies with an appearance that was suggestive of GBS were further processed to confirm the presence of GBS using general laboratory methods, which included: gram stain, catalase testing, and latex agglutination.

The testing of samples with Lim broth incubation periods per CDC guidelines included eighty-four (84) total runs with thirty-one (31) control pairs, all of which produced the expected positive/negative results. The invalid rate of the clinical prospective study samples with Lim broth incubation periods per CDC guidelines was 0.0%, zero out of four hundred and thirty-two (0/432). The results of the study are shown in Table 1.

| Simplexa™™ GBS

Direct Results	Culture Comparator Method
	Detected	Not Detected	Total
Detected	97	13b	110
Not Detected	3a	319	322
Total	100	332	432
	Sensitivity
97.0%(97/100)
95% CI: 91.5% to 99.0%	Specificity
96.1%(319/332)
95% CI: 93.4% to 97.7%

Table 1. Clinical Agreement Summary Results - Prospective Clinical Agreement Results Simplexa™ GBS Direct vs. Culture

ª 3/3 samples with discrepant results were negative when tested with an alternate FDA cleared NAAT.

b 11/13 samples with discrepant results were positive when tested with an alternate FDA cleared NAAT.

REPRODUCIBILITY

Three (3) investigative sites assessed the device's inter-day and inter/intra-assay reproducibility. Each of the laboratories tested Simplexa™ GBS Direct Positive Control, No Template Control (Lim Broth), and four (4) contrived samples in negative matrix. Two (2) strains of GBS were used in the study, BAA-22 and BAA-1138. The four (4) contrived samples consisted of a low positive (LP) at approximately 1.5 x LoD and a medium positive (MP) at approximately 3.5 x LoD for each GBS strain. The assays were performed in triplicate on five (5) different days. Each site had two (2) operators who each assayed the entire sample panel and Positive Control once per day, for a total of two (2) sets of data per day on a total of six (6) LIAISON® MDX instruments. The combined results for all sites are presented in Tables 2 and 3.

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Image /page/7/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 bold, dark blue font, with the word "Molecular" underneath in a lighter green font. The logo is clean and modern, suggesting a company involved in molecular diagnostics or biotechnology.

510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 5 of 19

| GBS
(FAM) | Site 1 | | | Site 2 | | | Site 4 | | | Total %
Agreement
with
Expected
Results | 95% CI |
|-----------------------------------------------|---------------------------------------------|--------------|-----------------------------------------------|-------------------------------------------|--------------|-----------------------------------------------|---------------------------------------------|--------------|-----|-----------------------------------------------------|--------------------|
| %
Agreement
with
Expected
Results | Avg
Ct | Total
%CV | %
Agreement
with
Expected
Results | Avg
Ct | Total
%CV | %
Agreement
with
Expected
Results | Avg
Ct | Total
%CV | | | |
| BAA-22 -
LP | 100.0%
(30/30) | 33.2 | 1.2 | 100.0%
(30/30) | 33.7 | 1.7 | 100.0%
(30/30) | 32.1 | 0.9 | 100.0%
(90/90) | 95.9% to
100.0% |
| BAA-22 -
MP | 100.0%
(30/30) | 32.6 | 1.1 | 100.0%
(30/30) | 31.6 | 1.3 | 100.0%
(30/30) | 31.9 | 1.1 | 100.0%
(90/90) | 95.9% to
100.0% |
| BAA-
1138 -
LP | 100.0%
(30/30) | 34.1 | 1.6 | 93.3%
(28/30) | 34.6 | 3.1 | 100.0%
(30/30) | 33.1 | 2.3 | 97.8%
(88/90) | 92.3% to
99.4% |
| BAA-
1138 -
MP | 100.0%
(30/30) | 32.7 | 1.2 | 100.0%
(30/30) | 33.1 | 1.4 | 100.0%
(30/30) | 31.9 | 1.5 | 100.0%
(90/90) | 95.9% to
100.0% |
| PC | 100.0%
(30/30) | 30.2 | 0.6 | 100.0%
(30/30) | 28.7 | 0.7 | 100.0%
(30/30) | 29.1 | 0.7 | 100.0%
(90/90) | 95.9% to
100.0% |
| NTC | 100.0%
(30/30) | NA | NA | 100.0%
(30/30) | NA | NA | 100.0%
(30/30) | NA | NA | 100.0%
(90/90) | 95.9% to
100.0% |
| Total
Agreement | 100.0% (180/180)
95% Cl: 97.9% to 100.0% | | | 98.9% (178/180)
95% Cl: 96.0% to 99.7% | | | 100.0% (180/180)
95% CI: 97.9% to 100.0% | | | 99.6% (538/540)
95% Cl: 98.7% to 99.9% | |

Table 2. Simplexa™ GBS Direct Reproducibility (FAM)

Table 3. Simplexa™ GBS Direct Internal Control Reproducibility (Q670)

	Site 1
Sample
DNA IC
(Q670)	%
Agreement
with
Expected
Results	Avg
Ct	Total
%CV	%
Agreement
with
Expected
Results	Avg
Ct	Total
%CV	%
Agreement
with
Expected
Results	Avg
Ct	Total
%CV	Agreement
with
Expected
Results	95% Cl
BAA-22 -
LP	100.0%
(30/30)	31.1	0.9	100.0%
(30/30)	31.1	0.7	100.0%
(30/30)	29.8	0.6	100.0%
(90/90)	95.9% to
100.0%
BAA-22 -
MP	100.0%
(30/30)	31.0	0.4	100.0%
(30/30)	30.3	0.6	100.0%
(30/30)	30.6	0.7	100.0%
(90/90)	95.9% to
100.0%
BAA-
1138 -
LP	100.0%
(30/30)	31.0	0.8	100.0%
(30/30)	31.1	0.4	100.0%
(30/30)	29.9	1.6	100.0%
(90/90)	95.9% to
100.0%
BAA-
1138 -
MP	100.0%
(30/30)	31.0	0.7	100.0%
(30/30)	31.1	0.5	100.0%
(30/30)	29.8	0.7	100.0%
(90/90)	95.9% to
100.0%
РС	100.0%
(30/30)	30.9	0.7	100.0%
(30/30)	30.2	0.4	100.0%
(30/30)	30.4	0.6	100.0%
(90/90)	95.9% to
100.0%
NTC	100.0%
(30/30)	31.0	0.5	100.0%
(30/30)	31.1	0.5	100.0%
(30/30)	30.0	0.9	100.0%
(90/90)	95.9% to
100.0%
Total
Agreement	100.0% (180/180)
95% Cl: 97.9% to 100.0%			100.0% (180/180)
95% Cl: 97.9% to 100.0%			100.0% (180/180)
95% Cl: 97.9% to 100.0%			100.0% (540/540)
95% Cl: 99.3% to
100.0%

8

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510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 6 of 19

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

The Limit of Detection (LoD) was determined for the Simplexa™ GBS Direct assay using quantified stocks of two (2) GBS strains (BAA-22 and BAA-1138) serially diluted into negative eighteen to twentyfour (18 to 24) hour Lim broth enrichments of vaginal/rectal specimen swabs obtained from antepartum women. The LoD was determined to be the lowest concentration that could be detected positive ≥ 95% of the time. The results are shown in Table 4.

Table 4. Simplexa™ GBS Direct Summary of Limit of Detection (LoD)

GBS strain	LoD Concentration (CFU/mL)
ATCC BAA-22
(serotype III)	80,000
ATCC BAA-1138
(serotype la)	30,000

ANALYTICAL REACTIVITY/CROSS REACTIVITY

Analytical Reactivity

The Simplexa™ GBS Direct assay was evaluated for analytical reactivity to an additional eighteen (18) GBS strains spiked at 2 x LoD into eighteen to twenty-four (18 to 24) Lim broth enrichments of vaginal/rectal specimen swabs obtained from antepartum women. The results are shown in Table 5. All eighteen (18) strains were detected.

Detected / # Tested |

|-----|---------------------------------|----------|--------------------------------------|--------------------------------------------------------------|
| 1 | ATCC BAA-1177 | Ia | 110,000 | 100% (3/3) |
| 2 | ATCC 51487 | Ib | 110,000 | 100% (3/3) |
| 3 | ATCC 27591 | Ic | 110,000 | 100% (3/3) |
| 4 | ATCC 12973 | II | 110,000 | 100% (3/3) |
| 5 | ATCC BAA-2675 | II | 110,000 | 100% (3/3) |
| 6 | ATCC BAA-1176 | III | 110,000 | 100% (3/3) |
| 7 | ATCC BAA-2674 | III | 110,000 | 100% (3/3) |
| 8 | ATCC 12403 | III | 110,000 | 100% (3/3) |
| 9 | ATCC 49446 | IV | 110,000 | 100% (3/3) |
| 10 | ATCC BAA-2673 | IV | 110,000 | 100% (3/3) |
| 11 | ATCC BAA-2672 | V | 110,000 | 100% (3/3) |
| 12 | ATCC BAA-611 | V | 110,000 | 100% (3/3) |
| 13 | ATCC BAA-2671 | VI | 110,000 | 100% (3/3) |
| 14 | ATCC BAA-2670 | VII | 110,000 | 100% (3/3) |
| 15 | ATCC BAA-2669 | VIII | 110,000 | 100% (3/3) |

Table 5. Simplexa™ GBS Direct Analytical Reactivity

9

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510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 7 of 19

Detected / # Tested |

|-----|---------------------------------|---------------|--------------------------------------|-------------------------------------------------------------|
| 16 | ATCC BAA-2668 | IX 1 | 110,000 | 100% (3/3) |
| 17 | ATCC 13813 | Non-Hemolytic | 110,000 | 100% (3/3) |
| 18 | ATCC BAA-2666 | Non-Hemolytic | 110,000 | 100% (3/3) |

Cross Reactivity (Analytical Specificity)

The Simplexa™ GBS Direct assay's analytical specificity was evaluated by testing the ability of the assay to exclusively identify GBS without any cross-reactivity to organisms that are closely related, or cause similar clinical symptoms or may be present in eighteen to twenty-four (18 to 24) hour Lim broth enrichments of vaginal/rectal specimen swabs obtained from antepartum women. Seventy-four (74) potential cross-reacting organisms were spiked into GBS negative enriched Lim broth and tested in triplicate. Four (4) organisms with low concentration stocks were additionally tested in silico. No cross reactivity was observed demonstrating the specificity of the Simplexa™ GBS Direct assay. The results are presented in Table 7.

| No. | Cross Reactant % Detection
(# Detected/ # Tested) |
|-----|---------------------- | 1 | Acinetobacter baumannii | 2 | Actinomyces israelii | 3 | Aerococcus viridans | 4 | Aeromonas hydrophila | 5 | Atopobium vaginae | 6 | Bacillus cereus | 7 | Bacteroides fragilis | 8 | Bifidobacterium adolescentis | 9 | Bifidobacterium breve | 10 | Blastocystis hominis | 11 | Brevibacterium linens | 12 | Campylobacter jejuni | 13 | Candida albicans | 14 | Candida glabrata | 15 | Candida parapsilosis | 16 | Candida tropicalis | 17 | Chlamydia trachomatis | 18 | Chromobacterium violaceum | 19 | Clostridium difficile | 20 | Clostridium perfringens | No. | Cross Reactant % Detection
(# Detected/ # Tested) |
| 21 | Corynebacterium genitalium | 22 | Cryptococcus neoformans | 23 | Enterobacter cloacae | 24 | Enterococcus avium | 25 | Enterococcus faecalis | 26 | Enterococcus faecium | 27 | Escherichia coli | 28 | Fusobacterium nucleatum | 29 | Gardnerella vaginalis* | 30 | Giardia lamblia | 31 | HSV-1 | 32 | HSV-2 | 33 | HIV-1 | 34 | HPV Genotype 18 | 35 | White Blood DNA) | 1 x 106 cells/mL | 36 | Human rotavirus | 37 | Klebsiella oxytoca | 38 | Klebsiella pneumoniae | 39 | Lactobacillus acidophilus | 40 | Lactobacillus crispatus | 41 | Lactobacillus | 42 | Lactobacillus iners | 43 | Lactobacillus jensensii | 44 | Mobiluncus curtisii | 45 | Mobiluncus mulieris | 46 | Mycoplasma genitalium | 47 | Mycoplasma hominis | 48 | Norovirus Genogroup GI* | 49 | Norovirus Genogroup GII* | 50 | Neisseria gonorrhoeae | 51 | Peptostreptococcus anaerobius | 52 | Porphyromonas asaccharolytica | 53 | Prevotella melaninogenica | No. | Cross Reactant % Detection
(# Detected/ # Tested) |
| 54 | Propionibacterium acnes | 55 | Proteus mirabilis | 56 | Pseudomonas aeruginosa | 57 | Salmonella enterica | 58 | Serratia marcescens | 59 | Shigella sonnei | 60 | Staphylococcus aureus | 61 | Staphylococcus epidermidis | 62 | Staphylococcus saprophyticus | 63 | Streptococcus anginosus | 64 | Streptococcus bovis | 65 | Streptococcus dysgalactiae | 66 | Streptococcus intermedius | 67 | Streptococcus mitis | 68 | Streptococcus oralis | 69 | Streptococcus pneumoniae | 70 | Streptococcus pyogenes | 71 | Streptococcus salivarius | 72 | Streptococcus sanguinis | 73 | Streptococcus suis | 74 | Streptococcus uberis | 75 | Trichomonas vaginalis* | 76 | Ureaplasma urealyticum | 77 | VZV | 78 | Yersinia enterocolitica | Tested Concentration | Expected Negative Results
--------------------|----------------------|--------------------------------------------------------------------|
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 cells/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 IFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| Tested Concentration | Expected Negative Results
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| NA | NA |
| 1 x 106 cells/mL | 0% (0/3) |
| 1 x 105 TCID50/mL | 0% (0/3) |
| 1 x 105 TCID50/mL | 0% (0/3) |
| 5 x 104 TCID50/mL * | 0% (0/3) |
| 1 x 105 IU/mL | 0% (0/3) |
Cells (Human genomic
| 0% (0/3) |
| 1 x 104 TCID50/mL * | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
delbrueckii subsp. lactis | 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 5 x 105 CCU/mL* | 0% (0/5)** |
| 1 x 106 CCU/mL | 0% (0/3) |
| NA | NA |
| NA | NA |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| Tested Concentration | Expected Negative Results
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
subsp enterica | 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |
| NA | NA |
| 1 x 106 CCU/mL | 0% (0/3) |
| 1 x 105 copies/mL | 0% (0/3) |
| 1 x 106 CFU/mL | 0% (0/3) |

Table 7. Simplexa™ GBS Direct Cross Reactivity (Analytical Specificity)

10

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510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 8 of 19

11

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510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 9 of 19

Tested in silico and no potential for cross-reaction with the Simplexa™ GBS Direct primers and probe observed. ** Tested in quintuplicate due to assay set up error.

NA = not available

INTERFERENCE

The performance of the Simplexa™ GBS Direct assay was evaluated with twenty-six (26) potentially interfering endogenous and exogenous substances that may be present in eighteen to twenty-four (18 to 24) hour Lim broth enrichments of vaginal/rectal specimen swabs obtained from antepartum women. All samples were prepared by spiking each potentially interfering substance into a baseline consisting of either the ATCC BAA-1138 or ATCC BAA-22 strain of GBS at a concentration of 2 x LoD in eighteen to twenty-four (18 to 24) hour Lim broth enrichments of vaginal/rectal specimen swabs obtained from antepartum women. Each interferent was spiked into the baseline sample and tested at the concentrations listed in Table 8. The results are presented in Table 8. No interference was observed at the concentrations indicated in Table 8.

12

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510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 10 of 19

	Potentially Interfering Substance	Active Ingredient	Interferent Concentration
1	Amniotic Fluid	NA	10% v/v
2	Antacids (Gaviscon)	Aluminum Hydroxide /
Magnesium Hydroxide	5 % v/v*
3	Antacids/Gastritis Medication	Calcium Carbonate	10 mg/mL
4	Anti-Diarrheal Medication
(Imodium)	Loperamide Hydrochloride	10% v/v
5	Anti-Diarrheal Medication (Pepto
Bismol)	Bismuth Subsalicylate	10% v/v
6	Anti-Fungal /Anti-Itch Vaginal	Nystatin	10 mg/mL**
7	Anti-Hemorrhoid
Creams/Ointments	Hydrocortisone	10 mg/mL
8	Anti-Hemorrhoid
Creams/Ointments	Phenylephrine HCl	10 mg/mL
9	Antimicrobials	Clotrimazole	10% w/v
10	Fecal Fat	Palmitic Acid	5 mg/mL*
11	Gastritis Medications (Nexium)	Esomeprazole Magnesium	0.31 mg/mL*
12	Laxatives	Sennosides	5 mg/mL*
13	Meconium	NA	0.31% w/v *
14	Mesalazine enema	Mesalazine	5 mg/mL*
15	Mineral Oil enema	Mineral Oil	10% v/v
16	Moist Towelettes	Benzalkonium Chloride	10% v/v
17	Moist Towelettes	Ethanol	10% v/v
18	Mucus	Immunoglobulin, Lysozyme,
Polymers	10% v/v
19	Non-Steroidal Anti-Inflammatory
Medications	Naproxen Sodium	3 mg/mL*
20	Radiology Oral Compounds	Barium Sulfate	5 mg/mL*
21	Seminal Fluid	NA	5% v/v*
	Potentially Interfering Substance	Active Ingredient	Interferent Concentration
22	Spermicidal Lubricant	Nonoxynol-9	10 mg/mL
23	Stool	NA	0.31% w/v*
24	Topical Products	Baby Powder	10 mg/mL
25	Topical Products	KY Jelly	10% w/v*
26	Whole Blood	Glucose, Hormones, Enzymes, Ions, Iron, etc.	10% v/v

Table 8. Simplexa™ GBS Direct Interference

13

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510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 11 of 19

False negative and/or Invalid results were obtained at higher concentrations

On initial testing 1 sample produced an EC505 error due to insufficient information to determine whether amplification occurred; the retest result was acceptable

INHIBITION BY OTHER MICROORGANISMS

The Simplexa™ GBS Direct assay was evaluated by testing the ability to identify GBS when other potentially inhibitory organisms were present at high concentrations. The panel of seventy-four (74) potentially inhibitory organisms were individually spiked into a pool with a low concentration of GBS BAA-22 or BAA-1138 strains at approximately 2 x LoD in Lim broth enrichments eighteen to twenty-four (18 to 24) hour of vaginal/rectal specimen swabs obtained from antepartum women. No false negative results were observed with GBS BAA-22 in the presence of any of the potentially interfering organisms or viruses. However, with GBS BAA-1138, at least one false negative result was obtained in the presence of each of the following species: Bacillus cereus, Candida parapsilosis, Clostridium perfringens, Peptostreptococcus anaerobius, Staphylococcus saprophyticus, Streptococcus pyogenes and Ureaplasma urealyticum (Table 9). Additional in silico analysis demonstrated no potential for crossreaction or interference with the Simplexa™ GBS Direct primers and probe by Norovirus genogroups I and II or Trichomonas vaginalis.

| No. | Organism | GBS Strain | Tested Concentration | % Detection
(# Detected/ # Tested) |
|-----|--------------------------------------------|------------|----------------------|---------------------------------------|
| 1 | Acinetobacter baumannii | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 2 | Actinomyces israelii | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 3 | Aerococcus viridans | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 4 | Aeromonas hydrophila | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| No. | Organism | GBS Strain | Tested Concentration | % Detection
(# Detected/ # Tested) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 5 | Atopobium vaginae | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 6 | Bacillus cereus | BAA-1138 | 1 x 106 CFU/mL | 88.9 %(8/9) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 7 | Bacteroides fragilis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 8 | Bifidobacterium adolescentis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 9 | Bifidobacterium breve | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 10 | Blastocystis hominis | BAA-1138 | 1 x 106 cells/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 cells/mL | 100% (3/3) |
| 11 | Brevibacterium linens | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 12 | Campylobacter jejuni | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 13 | Candida albicans | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 14 | Candida glabrata | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 15 | Candida parapsilosis | BAA-1138 | 1 x 106 CFU/mL | 77.8% (7/9) |
| No. | Organism | GBS Strain | Tested Concentration | % Detection
(# Detected/ # Tested) |
| 16 | Candida tropicalis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 17 | Chlamydia trachomatis | BAA-1138 | 1 x 106 IFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 IFU/mL | 100% (3/3) |
| 18 | Chromobacterium violaceum | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 19 | Clostridium difficile | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 20 | Clostridium perfringens | BAA-1138 | 1 x 106 CFU/mL | 88.9% (8/9) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 21 | Corynebacterium genitalium | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 22 | Cryptococcus neoformans | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 23 | Enterobacter cloacae | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 24 | Enterococcus avium | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 25 | Enterococcus faecalis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 26 | Enterococcus faecium | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| No. | Organism | GBS Strain | Tested Concentration | % Detection
(# Detected/ # Tested) |
| 27 | Escherichia coli | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| 28 | Fusobacterium nucleatum | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| 29 | Giardia lamblia | BAA-22 | 1 x 106 cells/mL | 100% (3/3) |
| | | BAA-1138 | 1 x 106 cells/mL | 100% (3/3) |
| 30 | HIV-1 | BAA-22 | 5 x 104 TCID50/mL | 100% (3/3) |
| | | BAA-1138 | 5 x 104 TCID50/mL | 100% (3/3) |
| 31 | HPV Genotype 18 | BAA-22 | 1 x 105 IU/mL | 100% (3/3) |
| | | BAA-1138 | 1 x 105 IU/mL | 100% (3/3) |
| 32 | HSV-1 | BAA-22 | 1 x 105 TCID50/mL | 100% (3/3) |
| | | BAA-1138 | 1 x 105 TCID50/mL | 100% (3/3) |
| 33 | HSV-2 | BAA-22 | 1 x 105 TCID50/mL | 100% (3/3) |
| | | BAA-1138 | 1 x 105 TCID50/mL | 100% (3/3) |
| 34 | Human DNA (White Blood
Cells) | BAA-22 | 1 x 106 WBC/mL | 100% (3/3) |
| | | BAA-1138 | 1 x 106 WBC/mL | 100% (3/3) |
| 35 | Human rotavirus | BAA-22 | 1 x 104 TCID50/mL | 100% (3/3) |
| | | BAA-1138 | 1 x 104 TCID50/mL | 100% (3/3) |
| 36 | Klebsiella oxytoca | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| No. | Organism | GBS Strain | Tested Concentration | % Detection
(# Detected/ # Tested) |
| 37 | Klebsiella pneumoniae | BAA-1138 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| 38 | Lactobacillus acidophilus | BAA-1138 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| 39 | Lactobacillus crispatus | BAA-1138 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| 40 | Lactobacillus delbrueckii
subsp. lactis | BAA-1138 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| 41 | Lactobacillus iners | BAA-1138 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| 42 | Lactobacillus jensensii | BAA-1138 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| 43 | Mobiluncus curtisii | BAA-1138 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| 44 | Mobiluncus mulieris | BAA-1138 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| 45 | Mycoplasma genitalium | BAA-1138 | $5 x 10^5$ CCU/mL | 100% (3/3) |
| | | BAA-22 | $5 x 10^5$ CCU/mL | 100% (3/3) |
| 46 | Mycoplasma hominis | BAA-1138 | $1 x 10^6$ CCU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CCU/mL | 100% (3/3) |
| 47 | Neisseria gonorrhoeae | BAA-1138 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| | | BAA-22 | $1 x 10^6$ CFU/mL | 100% (3/3) |
| No. | Organism | GBS Strain | Tested Concentration | % Detection
(# Detected/ # Tested) |
| 48 | Norovirus genogroup I* | NA | NA | NA |
| 49 | Norovirus genogroup II* | NA | NA | NA |
| 50 | Peptostreptococcus
anaerobius | BAA-1138 | 1 x 106 CFU/mL | 88.9% (8/9) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 51 | Porphyromonas
asaccharolytica | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 52 | Prevotella melaninogenica | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 53 | Propionibacterium acnes | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 54 | Proteus mirabilis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 55 | Pseudomonas aeruginosa | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 56 | Salmonella enterica subsp
enterica | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 57 | Serratia marcescens | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 58 | Shigella sonnei | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 59 | Staphylococcus aureus | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| No. | Organism | GBS Strain | Tested Concentration | % Detection
(# Detected/ # Tested) |
| 60 | Staphylococcus epidermidis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 61 | Staphylococcus saprophyticus | BAA-1138 | 1 x 106 CFU/mL | 77.8% (7/9) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 62 | Streptococcus anginosus | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 63 | Streptococcus bovis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 64 | Streptococcus dysgalactiae | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 65 | Streptococcus intermedius | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 66 | Streptococcus mitis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 67 | Streptococcus oralis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 68 | Streptococcus pneumoniae | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 69 | Streptococcus pyogenes | BAA-1138 | 1 x 106 CFU/mL | 88.9% (8/9) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 70 | Streptococcus salivarius** | BAA-1138 | 1 x 106 CFU/mL | 100% (6/6) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (6/6) |
| No. | Organism | GBS Strain | Tested Concentration | % Detection
(# Detected/ # Tested) |
| 71 | Streptococcus sanguinis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 72 | Streptococcus suis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 73 | Streptococcus uberis | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |
| 74 | Trichomonas vaginalis* | NA | NA | NA |
| 75 | Ureaplasma urealyticum | BAA-1138 | 1 x 106 CCU/mL | 77.8% (7/9) |
| | | BAA-22 | 1 x 106 CCU/mL | 100% (3/3) |
| 76 | VZV | BAA-1138 | 1 x 105 copies/mL | 100% (3/3) |
| | | BAA-22 | 1 x 105 copies/mL | 100% (3/3) |
| 77 | Yersinia enterocolitica | BAA-1138 | 1 x 106 CFU/mL | 100% (3/3) |
| | | BAA-22 | 1 x 106 CFU/mL | 100% (3/3) |

Table 9. Simplexa™ GBS Direct Microbial Inhibition

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510(k) Summary

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

510(k) Summary

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Image /page/16/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo consists of a DNA helix graphic on the left, with the text "DiaSorin" in dark blue to the right of the helix. Below "DiaSorin" is the word "Molecular" in green.

510(k) Summary

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Image /page/17/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 dark blue, and below that, the word "Molecular" is written in light green.

510(k) Summary

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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 dark blue on the top line and "Molecular" in light green on the bottom line.

510(k) Summary

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Image /page/19/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 on the top line and "Molecular" in light green on the bottom line.

510(k) Summary

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Image /page/20/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 the word "Molecular" underneath in a lighter green color. The logo is clean and modern, suggesting a company involved in molecular diagnostics or biotechnology.

510(k) Summary

Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 18 of 19

Tested in silico and no potential for cross-reaction or interference with the Simplexa™ GBS Direct primers and probe observed

** Streptococcus salivarius was inadvertently tested twice.

NA = not available

CARRY-OVER CONTAMINATION

Amplification carry-over for the Simplexa™ assays has been assessed. The study was performed by testing alternating high positive and negative samples on each disc. No evidence of carry-over contamination was observed.

EXPECTED VALUES

The prevalence of GBS as determined by the Simplexa™ GBS Direct assay in a multi-site clinical study with prospectively collected specimens is shown in Table 10.

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Image /page/21/Picture/0 description: The image shows the logo for DiaSorin Molecular. The logo features a stylized DNA helix in shades of green and blue on the left. To the right of the helix are the words "DiaSorin" in dark blue on the top line and "Molecular" in light green on the bottom line.

510(k) Summary Simplexa™ GBS Direct Catalog No. MOL3550 Simplexa™ GBS Positive Control Pack Catalog No. MOL3560 October 25, 2018 Page 19 of 19

| Site ID | Total | Simplexa™ GBS Direct
Positive | GBS Prevalence |
|---------|-------|----------------------------------|----------------|
| 01 | 168 | 49 | 29.2% |
| 05 | 208 | 44 | 21.2% |
| 06 | 56 | 17 | 30.4% |
| All | 432 | 110 | 25.5% |

Table 10. Expected Values per Site

The expected values for GBS per site varied between 21.2% and 30.4%. The overall prevalence for all sites was 25.5% one hundred and ten of four hundred and thirty-two (110/432) by the Simplexa™ GBS Direct assay and 23.1% one hundred of four hundred and thirty-two (100/432) as determined by a culture reference method.

CONCLUSION

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.