The AmpliVue® GBS Assay is a qualitative in vitro diagnostic test for the rapid detection of Group B Streptococcus from vaginal/rectal swabs from antepartum women following 18 to 24 hours of incubation in an LIM enrichment broth culture. The AmpliVue" GBS Assay utilizes helicase-dependent amplification (HDA) of the thiolase (atoB) gene sequence and a self-contained disposable amplification detection device that allows for manual evaluation of assay results. Results can be used as an aid in determining the colonization status of antepartum women.

The AmpliVue GBS Assay does not provide susceptibility results. Culture isolates are needed for performing susceptibility testing as recommended for penicillin-allergic women.

The AmpliVue® GBS Assay is intended for use in hospital, reference or state laboratory settings. The device is not intended for point-of-care use.

Device Description

The AmpliVue® GBS Assay combines simple sample processing, an isothermal amplification technology named helicase-dependent amplification (HDA), and a selfcontained disposable amplicon detection device, for the detection of Group B Streptococcus from vaginal /rectal swabs following 18 to 24 hours incubation in Lim enrichment broth culture.

A small amount of cultured specimen is transferred into a dilution tube. The diluted sample culture is then transferred into a lysis tube, and the cells are lysed by simple heat treatment. After heat treatment, an aliquot of the lysed sample is added to a reaction tube containing a lyophilized mix of HDA reagents including primers specific for the amplification of the thiolase (atoB) gene. The rationale behind the selection of this particular target sequence were: 1) a BLAST search of this gene resulted in no thiolase with significant homology in species other than those belonging to S. agalactiae; 2) the gene is conserved in GBS based on the recent GBS Genome project. The assay also includes a process control that monitors sample processing, confirms the integrity of the assay reagents and cassette detection, and assays for HDA-inhibitors that may be present within a broth culture. After completion of the HDA reaction, the reaction tube is transferred to a cassette for rapid detection with test result displayed as test and/or control lines in the window of the cassette.

AI/ML Overview

Here's a breakdown of the acceptance criteria and study details for the AmpliVue® GBS Assay, based on the provided 510(k) summary:

Acceptance Criteria and Device Performance

1. Table of Acceptance Criteria and Reported Device Performance:

The document doesn't explicitly state "acceptance criteria" in a separate table with target values for clinical performance. However, it presents the clinical sensitivity and specificity results. For the purpose of this response, we'll interpret the reported clinical performance metrics as reflecting the achieved acceptance criteria. The predicate device's performance is also included for context, as it would have likely informed the expected performance of the new device.

Performance Metric	Acceptance Criteria (Implied / Comparator)	Reported Device Performance (AmpliVue® GBS Assay)
Clinical Sensitivity	N/A (Predicate: 97.4% (95% CI: 91.9 - 99.0%))	99.5% (95% CI: 96.9 - 100%)
Clinical Specificity	N/A	92.7% (95% CI: 90.5 - 94.3%)
LoD (Limit of Detection)	N/A (Lowest concentration for 95% positivity)	1.39 x 10^6 CFU/mL (based on highest value among 6 strains)
Reproducibility	Acceptable (no significant differences between users, sites, lots)	Achieved (100% agreement for low positive, moderate positive, negative controls; 78% for GBS High Negative overall)
Cross-reactivity	No cross-reactivity with tested organisms/viruses at specified levels	Achieved (None of 91 organisms/viruses, nor human genomic DNA, cross-reacted)
Interference	No interference with tested substances at specified concentrations	Achieved (None of 34 substances interfered)
Analytical Reactivity (Inclusivity)	Detection of various GBS strains at 1x LoD	Achieved (All 12 tested strains detected at 1x LoD)

Note: The document explicitly states "Reproducibility studies are acceptable" without providing a numerical acceptance criterion. For clinical sensitivity and specificity, no explicit numerical acceptance criterion is provided for the AmpliVue® GBS Assay in this summary. Instead, its performance is presented, and it is also compared to the predicate device's clinical sensitivity.

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

Sample Size for Clinical Study (Test Set): 911 specimens were initially collected. After accounting for invalid samples (3 initially, 3 remaining invalid after repeat testing), the clinical performance was calculated based on 908 specimens.
Data Provenance:
- Country of Origin: United States (four distinct geographical sites).
- Retrospective or Prospective: Prospective study conducted from July to November 2013.
Sample Size for Analytical Studies (LoD, Cross-reactivity, Interference, Inclusivity):
- LoD: For each of the six GBS strains, 20 replicates were tested, across 5 experiments of 14 assays per strain.
- Cross-reactivity: Two GBS strains (BAA-611 and SS617) tested near LoD alongside a panel of 91 organisms (76 bacteria, 3 yeast, 11 viruses, 1 parasite) and human genomic DNA.
- Interference: Two GBS strains (BAA-611 and SS617) tested near LoD with a panel of 34 substances.
- Inclusivity: Twelve additional strains of Streptococcus agalactiae were tested as three replicates each, near the LoD.

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

The document does not explicitly state the "number of experts" used to establish the ground truth or their specific qualifications (e.g., "radiologist with 10 years of experience").

However, the ground truth for the clinical study was established by bacterial culture. This implies that laboratory professionals (e.g., microbiologists, medical technologists) followed established protocols for bacterial culture, which serves as the "gold standard" or ground truth for GBS detection in this context.

For discordant specimens (AmpliVue Positive/Bacterial Culture Negative, and AmpliVue Negative/Bacterial Culture Positive), an FDA-cleared molecular device for the detection of GBS was used for further testing. This implies that the results from this FDA-cleared molecular device were leveraged to refine the understanding of the true GBS status in these cases.

4. Adjudication Method for the Test Set:

Based on the information provided, the adjudication method for the clinical test set seems to be:

Primary Ground Truth: Bacterial culture.
Discordant Analysis: For specimens where the AmpliVue® GBS Assay and bacterial culture disagreed, an FDA-cleared molecular device for the detection of GBS was used to re-evaluate the specimen. The results from this molecular device were then factored into the final assessment of the true positive/negative status for the discordant samples, as indicated by the footnotes in the clinical performance table. For example, of the 52 AmpliVue Positive/Bacterial Culture Negative samples, 39 were determined positive by the FDA-cleared molecular device. Similarly, the 1 AmpliVue Negative/Bacterial Culture Positive sample was found negative by the FDA-cleared molecular device.

This method resembles a tie-breaker or reference method adjudication rather than an expert consensus voting system.

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, a Multi Reader Multi Case (MRMC) comparative effectiveness study was not done.

This device is an in vitro diagnostic (IVD) assay for direct detection of a pathogen, not an AI-powered diagnostic imaging tool that assists human readers. Therefore, the concept of "human readers improve with AI vs without AI assistance" is not applicable to this type of device. The assay results are read manually (colored lines visible to the naked eye) by a single operator for each test, rather than a panel of human readers interpreting complex medical images.

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

The performance reported in the clinical study (Clinical Sensitivity and Specificity) is a standalone performance of the AmpliVue® GBS Assay. The assay produces a direct result (test and/or control lines in the cassette window visible to the naked eye) that is interpreted by an operator. While an operator reads the result, it's not a "human-in-the-loop" scenario in the sense of a subjective interpretation or decision-making process that would significantly alter the algorithm's output. The output of the HDA reaction and detection cassette directly provides the positive/negative result.

7. The Type of Ground Truth Used (expert consensus, pathology, outcomes data, etc.):

The primary ground truth used for the clinical study was bacterial culture, which is the conventional gold standard for confirming GBS colonization. For discordant results, further confirmation was sought using an FDA-cleared molecular device for the detection of GBS.

8. The Sample Size for the Training Set:

The document does not provide information on a specific "training set" sample size. This is common for molecular diagnostic assays that detect specific gene sequences. The assay's "training" largely relies on the initial design and optimization of primers, probes, and reaction conditions to target the specific atoB gene sequence of S. agalactiae based on existing genomic knowledge. The clinical studies and performance evaluations described are validation studies, not "training" of a machine learning model.

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

As there is no explicit "training set" in the context of machine learning, the question of how its ground truth was established is not directly applicable.

However, the design of the assay (selection of the atoB gene as a target) was based on:

BLAST search indicating no significant homology in non-S. agalactiae species.
Conservation of the gene in GBS based on the recent GBS Genome project.

These bioinformatics and genomic reference data can be considered the "ground truth" that informed the initial assay design and target selection. The analytical and clinical validation studies then confirm that this design effectively detects GBS in real-world samples.

Summary

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K133503

Quidel Corporation

AmpliVue® GBS Assay Page 1 of 15

510(k) Summary

Applicant:

DEC 2 0 2013

Quidel Corporation 10165 McKellar Court San Diego, California 92121 Telephone: 858-552-7910 Fax: 858-646-8045

Contact Information:

Ronald H. Lollar, Senior Director Clinical and Quality Affairs 1055 East State Street Suite 100 Athens, Ohio 45701 740-589-3300 - Corporate number 740-589-3373 – Desk phone 740-593-8437 - Fax lollar@dhiusa.com

Date of preparation of 510(k) summary:

December 19, 2013

A. 510(k) Number:

K133503

B. Purpose for Submission:

To obtain substantial equivalence for the AmpliVue® GBS Assay

C. Measurand:

Thiolase (atoB) gene of S. agalactiae (Group B Streptococcus)

D. Type of Test:

Helicase-dependent amplification (HDA)

E. Applicant:

Quidel Corporation

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F. Proprietary and Established Names:

AmpliVue® GBS Assay

G. Regulatory Information:

NJR - Nucleic AcidAmplification AssaySystem, Group BStreptococcus,Direct Specimen	Class I (Non-exempt)	21 CFR 866.3740 - Streptococcus spp.Serological Reagents	Microbiology (83)

H. Intended Use:

1. Intended Use(s):

The AmpliVue GBS Assay is a qualitative in vitro diagnostic test for the rapid detection of Group B Streptococcus from vaginal/rectal swabs from antepartum women following 18 to 24 hours of incubation in an LIM enrichment broth culture. The AmpliVue" GBS Assay utilizes helicase-dependent amplification (HDA) of the thiolase (atoB) gene sequence and a self-contained disposable amplification detection device that allows for manual evaluation of assay results. Results can be used as an aid in determining the colonization status of antepartum women.

The AmpliVue GBS Assay does not provide susceptibility results. Culture isolates are needed for performing susceptibility testing as recommended for penicillin-allergic women.

The AmpliVue® GBS Assay is intended for use in hospital, reference or state laboratory settings. The device is not intended for point-of-care use.

2. Indication(s) for Use:

The AmpliVue® GBS Assay is a qualitative in vitro diagnostic test for the rapid detection of Group B Streptococcus from vaginal/rectal swabs from antepartum women following 18 to 24 hours of incubation in an LIM enrichment broth culture. The AmpliVue GBS Assay utilizes helicase-dependent amplification (HDA) of the thiolase (atoB) gene sequence and a self-contained disposable amplification detection device that allows for manual evaluation of assay results. Results can be used as an aid in determining the colonization status of antepartum women.

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The AmpliVue GBS Assay does not provide susceptibility results. Culture isolates are needed for performing susceptibility testing as recommended for penicillin-allergic women.

The AmpliVue® GBS Assay is intended for use in hospital, reference or state laboratory settings. The device is not intended for point-of-care use.

3. Special conditions for use statement(s):

For in vitro diagnostic use only .
For prescription use only .

4. Special instrument requirements:

None

Device Description: I.

Materials Provided:

· 16 Tests per Kit

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Symbol	Component	Quantity	Storage
1	Detection Cassettes	16/kit	2° to 30°C
2	Dilution Buffer	16 tubes/kit 0.35 mL	2° to 8°C
3	Lysis Buffer	16 tubes/kit 0.35 mL	2° to 8°C
4	Reaction Tubes	16 tubes/kit	2° to 8°C
5	Amplicon Cartridge	16/kit	2° to 30°C

Materials required but not provided:

External controls for Group B Streptococcus (e.g. AmpliVue® GBS Control Set, . which contains positive and negative controls, serves as an external processing and extraction control)
Sterile DNAse-free filter-blocked or positive displacement micropipettor tips .
. Micropipettor
Stopwatch or timer .
. Scissors or a blade
Micro tube tray .
Heat block capable of 95° C ± 2° C temperature .
Heat block with heated lid capable of 64° ± 2° C temperature .
. Thermometer

J. Substantial Equivalence Information:

1. Predicate device name(s):
  illumigene® GBS Assay, external control kit (Meridian Bioscience, Inc.)
1. Predicate 510(k) number(s):
  K112125

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and the same of the same of the same of the same of the same of the same of the states of the states of the states of the states of the states of the states of the states of

510(k) Summary

Comparison with predicate: 3.

Similarities
Item	Device (K133503)	Predicate (K112125)
Intended Use	Qualitative in vitro diagnostic testfor the rapid detection of Group BStreptococcus from vaginal/rectalswabs from antepartum womenfollowing 18 to 24 hours ofincubation in enriched Lim brothculture	Same
Sample Types	Enriched Lim broth cultures ofVaginal/Rectal swab specimens	Same
Sample Heat Lysis	Manual	Same
Self-Contained System Assayafter sample preparation	Yes	Same

Differences
Item	Device (K133503)	Predicate (K112125)
Sample Type EnrichmentCulture	Lim broth cultures, only	Trans Vag and Lim broth
DNA Amplification Technology(HDA)	Helicase-dependent amplification(HDA)	Loop-mediated isothermal DNAamplification (LAMP)
Target Sequence Detected	Thiolase (atoB) gene	213 base pair (bp) sequenceresiding in the 593-805 bp regionof S. agalactiae genomeSegment 3
Detection Technique	Manual	Automated
Instrument	None	illumipro-10
Testing Time	75 - 90 minutes	60 minutes
Clinical Sensitivity	99.5% (95% CI: 96.9-100%)	97.4% (95% CI: 91.9 - 99.0%)

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	Differences
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K. Standard/Guidance Document Referenced (if applicable):

Not applicable

L. Test Principle:

Following a specimen lysis step, amplification of the atoB gene occurs if GBS is present in the HDA reaction. Competitive amplification of the process control DNA also takes place unless amplification inhibitory substances are present or the sample processing fails. The HDA reaction is asymmetric so that an excess of single stranded DNA is formed from a biotinylated primer present within the reaction mix.

The capture probes for each amplicon bind to the corresponding biotinylated single-stranded DNA and this dual-labeled probe-amplicon hybrid are then detected by using a proprietary detection cassette. The bottom line captures the FITC-labeled probe-target amplicon and the top line captures the DNP-labeled probe-control amplicon. The biotin label attracts the streptavidin-conjugated color particles for visualization and the test result is shown as colored lines visible to the naked eye. The self-contained cassette is comprised of two individual components: an amplicon cartridge that holds the running buffer and a single 0.2-mL thin wall reaction tube containing the amplified product; and the detection chamber which houses the amplicon Quidel Corporation cartridge and a vertical-flow DNA detection strip embedded into the cassette. The DNA detection strip is coated with an anti-FITC antibody and an anti-DNP antibody that serve as the GBS test (T) line and the control (C) line respectively in the assay. A razor blade and a plastic pin located at the bottom of the detection chamber open the HDA reaction tube and the running buffer bulb when the handle of the detection chamber is closed. The mixture flows through a fiberglass paper connected to the DNA detection strip that is attached with a fiberglass pad pre-loaded with streptavidin-conjugated color particles for color visualization. Detection of GBS is reported when the T line is visible through the detection window of the cassette. No detection of GBS is reported when only the C line is displayed. The assay is regarded as unresolved when neither line is displayed.

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M. Performance Characteristics:

1. Analytical performance:
- a. Precision/Reproducibility:

Reproducibility

In order to confirm the reproducibility of the AmpliVue® GBS Assay, a blinded and randomized study panel containing Streptococcus agalactiae negative and positive samples were tested at three test sites (one in-house laboratory and two clinical sites). Each site tested a reproducibility panel and Assay Controls for five days in triplicate. Testing was done by two operators at each site. Each operator ran the panel once a day using one lot of AmpliVue® GBS Assay. A total of 540 specimens were tested (including controls). The AmpliVue® GBS Assay generated the following reproducible results in this study.

	Site #1		Site #2		Site #3
Category	#expectedresults/#tested	%Agreement	#expectedresults/#tested	%Agreement	#expectedresults/#tested	%Agreement	Overall Results		95%ConfidenceInterval
GBS HighNegative	18/30	60%	24/30	80%	28/30	93%	70/90	78%	68% -85%
GBS LowPositive	30/30	100%	30/30	100%	30/30	100%	90/90	100%	95% -100%
GBSModeratePositive	30/30	100%	30/30	100%	30/30	100%	90/90	100%	95% -100%
GBSNegative	30/30	100%	30/30	100%	30/30	100%	90/90	100%	95% -100%
GBSPositiveControl	30/30	100%	30/30	100%	30/30	100%	90/90	100%	95% -100%
GBSNegativeControl	30/30	100%	30/30	100%	30/30	100%	90/90	100%	95% -100%

The results suggest that there are no significant differences between different users, different sites and different lots in different days. Reproducibility studies are acceptable.

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b. Linearity/assay reportable range:
Not applicable - This assay is qualitative.
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Traceability:

Not applicable. This assay is qualitative.

Specimen Stability:

A study was performed to determine the stability of samples in Lim Broth that are used in the AmpliVue GBS Assay. A freshly grown stock of GBS bacteria of known titer was used to spike a pooled negative clinical sample in Lim Broth. The spiked samples were stored at 4°C or 25°C and tested at 2h. 8h, 24h and 25h time points prior to being tested in the AmpliVue® GBS Assay.

Based on this study, clinical samples are expected to be stable at both 25°C and 4°C for up to 25 hours.

Controls:

Controls (AmpliVue GBS Control Set, which contains positive and negative controls, serves as an external processing and extraction control) were run on the AmpliVue® GBS Assay each day of testing. These controls are described as follows:

a. The process control is used to monitor sample processing, to detect HDA inhibitory specimens and to confirm the integrity of assay reagents and cassette detection. The process control is included in the Dilution Buffer tube.
The external positive control may be treated as a patient specimen. The b. control should be sampled and tested as if it were a broth culture and processed as described in the Assay Procedure. The external positive control is intended to monitor substantial reagent and cassette failure.
The external negative control may be treated as a patient specimen. The C. control should be sampled and tested as if it were a broth culture and processed as described in the Assay Procedure. The external negative control is used to detect reagent or environmental contamination (or carry-over) by GBS DNA or amplicon.

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d. Detection limit:

The analytical sensitivity (limit of detection or LoD) of the AmpliVue® GBS Assay was determined using quantified (CFU/mL) cultures of six Streptococcus agalactiae strains serially diluted in a Lim broth. The LoD is defined as the lowest concentration at which 95% of all replicates tested positive.

The GBS bacterial strains were freshly grown. The cell density of these bacterial suspensions was estimated using the McFarland standard OD reading. After a cell suspension of 0.25 McFarland units were established, the bacteria were serially diluted in Lim broth to densities ranging from 3x to 0.3x LoD levels based on preliminary studies.

Each test dilution was run as 20 replicates in the AmpliVue® assay and plated on 20 TSA + 5% blood plates. The study was performed in 5 experiments of 14 assays per strain. For each experiment. 4 replicates of each of the three Lim broth bacterial dilutions were performed, along with a positive and a negative run control. All five experiments of the LoD study for each strain were completed within 8 hours. The stocks of cells were stored on ice or at 4 ℃ when not in use.

The highest dilution where at least 19 of 20 replicates show detection of GBS (95% positivity) was assigned the Limit of Detection of the strain. The CFU/mL was calculated based on the average bacterial plate count of the dilution.

The LoD for the six strains tested ranged from 1.24 x 10° to 1.39 x 10° CFU/mL. Based on this data the reported LoD for the AmpliVue® GBS Assay is the highest value of the six strains: 1.39 x 106 CFU/mL.

Strain	CFU/mL
BAA-611	1.39 x 106
SS617	6.96 x 105
SS618	6.02 x 105
SS619	5.64 x 105
ATCC 12403	8.84 x 105
SS700	1.24 x 105

LoD Results

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e. Analytical specificity:
Cross Reactivity:

A study was conducted using two strains of Streptococcus agalactiae (BAA-611 and SS617) tested near LoD to evaluate the AmpliVue® GBS Assay for potential interference using a panel of 91 organisms (76 bacteria, three yeast, 11 viruses and a parasite listed in the tables below) potentially found in vaginal/rectal samples cross-react with the AmpliVue® GBS Assay. Human genomic DNA was also evaluated for cross-reactivity. The microorganisms were tested above clinically relevant levels (bacteria ≥ 1 x 106 CFU/mL, viruses ≥ 1 x 105 TCID50/mL).

Bacteria
Abiotrophia defective	Enterobacter cloacae	Salmonella choleraesius(typhimurium)
Acinetobacter baumanii	Enterococcus faecalis	Salmonella enterica arizonae
Aeromonas hydrophila	Enterococcus faecium	Salmonella enterica entericaSerovar Typhimurium
Alcaligenes faecalis faecalis	Escherichia coli	Salmonella enterica indica
Bacillus cereus	Escherichia fergusonii	Serratia liquefaciens
Bacillus subtilis	Gardnerella vaginalis	Serratia marcescens
Bacteroides fragilis	Helicobacter pylori	Shigella boydii
Bifidobacterium adolescentis	Klebsiella oxytoca	Shigella flexneri
Campylobacter fetus	Klebsiella pneumoniae	Shigella sonnei
Campylobacter hyointestinalis	Lactobacillus acidophilus	Staphylococcus aureus
Campylobacter jejuni	Legionella pneumophila	Staphylococcus epidermidis
Chlamydia trachomatis	Listeria monocytogenes	Stenotrophomonas maltophila
Citrobacter freundii	Mobiluncus mulieris	Streptococcus mutans
Clostridium bifermentans	Moraxella cartarrhalis	Streptococcus pyogenes
Clostridium butyricum	Morganella morganii	Streptococcus bovis

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Bacteria
Clostridium difficile	Neisseria gonorrhoeae	Streptococcus dysgalactiae
Clostridium haemolyticum	Peptostreptococcus anaerobius	Streptococcus gordonii
Clostridium novyi	Pleisiomonas shigelloides	Streptococcus intermedius
Clostridium orbiscindens	Porphyromonasasaccharolytica	Streptococcus mitis
Clostridium perfringens	Prevotella melaninogenica	Streptococcus oralis
Clostridium septicum	Proteus mirabilus	Streptococcus pneumonia
Clostridium sordellii	Providencia alcalifaciens	Streptococcus salivarius
Clostridium sporogenes	Pseudomonas aeruginosa	Streptococcus suis
Edwardsiella tarda	Pseudomonas fluorescens	Streptococcus uberis
Enterobacter aerogenes	S. dysgalactiae equisimilis	Vibrio parahaemolyticus
	.	Yersinia enterocolitica

Yeast
Candida albicans	Candida glabrata	Candida parapsilosis

Viruses
Adenovirus	Enterovirus	Norovirus
CMV	HPV-16	Rotavirus
Coxsackie virus	HSV1 (Macintyre)	VZV
Echovirus	HSV2 (G)

Parasite

Trichomonas vaginalis

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None of the organisms or viruses tested above cross-reacts with the performance of the AmpliVue® GBS Assay. In addition, human genomic DNA does not interfere with the performance of the AmpliVue® GBS Assay.

Interference:

A study was conducted using two strains of Streptococcus agalactiae (BAA-611 and SS617) tested near LoD to evaluate the AmpliVue GBS Assay for potential interference using a panel consisting of 34 substances (listed in the table below) found in vaginal/rectal specimens. Substances were introduced into the assay dilution tubes at concentrations which were medically relevant. Each of the strains was tested for each substance. None of the substances tested were found to interfere with the AmpliVue® GBS Assay.

Substance Name	ConcentrationTested	GBSResult	Substance Name	ConcentrationTested	GBSResult
ImodiumAD®(Loperamide HCl)	1.5 µg/mL	Positive	Witch Hazel	0.15%	Positive
Mucin	4.5 µg/mL	Positive	Benzalkonium Cl	0.00018%	Positive
Barium sulfate	7.5 µg/mL	Positive	Fecal Sugar - Dextrose	1.5 µg/mL	Positive
Fecal Fat - Palmitic Acid	1.95 µg/mL	Positive	Human Serum Albumin	15 µg/mL	Positive
Mylanta®Mg(OH)2	0.15 µg/mL	Positive	Mineral Oil	0.015% v/v	Positive
Hemorrhoidal cream(Phenylephrine HCl)	30 µg/mL	Positive	Triclosan	0.001%	Positive
Hemoglobin	4.8 µg/mL	Positive	Hemorrhoidal cream(Target Brand Cream)	0.015% ofswab	Positive
Mylanta® Al(OH)3	0.15 µg/mL	Positive	Anusol®(Hydrocortisone Cream)	0.015% ofswab	Positive
Prilosec® (EsomeprazoleMagnesium Hydrate)	0.75 µg/mL	Positive	Desitin®(Zinc Oxide)	0.015% ofswab	Positive
Tums® (CalciumCarbonate)	0.75 µg/mL	Positive	KY Jelly	0.015% ofswab	Positive

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Substance Name	ConcentrationTested	GBSResult	Substance Name	ConcentrationTested	GBSResult
Fecal Fat - Stearic Acid	39 µg/mL	Positive	Petroleum Jelly	0.015% ofswab	Positive
Kaopectate® (BismuthSubsalicylate)	1.3 µg/mL	Positive	Condom Swab(Nonoxynol-9)	0.015% ofswab	Positive
Tagamet® (Cimetidine)	0.75 µg/mL	Positive	Body Powder	0.015% ofswab	Positive
Ethanol	0.015% v/v	Positive	Meconium	0.015% ofswab	Positive
Miconazole Nitrate Salt	0.003%	Positive	Amniotic Fluid	0.0750%	Positive
Nystatin	15 USP U/mL	Positive	Stool	0.015%	Positive
Urine	0.05%	Positive	Whole Blood	0.006%	Positive

Analytical Reactivity (Inclusivity):

The reactivity of the AmpliVue® GBS Assay was evaluated against an additional twelve strains of Streptococcus agalactiae. The testing was performed near the level of detection for the assay (1x LoD).

Each strain was tested as three replicates in the AmpliVue® GBS Assay. The study was performed in multiple experiments of no more than 14 assays per experiment. For each experiment, three replicates of up to four strains were performed, along with a positive and a negative run control. All twelve strains were detected by the AmpliVue® GBS Assay in this study at a LoD of 1084 CFU/assay (1.39 x 106 CFU/mL).

Inclusivity Results
Bacterial Strain	Serotype	Vendor
ATCC 12973	II	ATCC
CCUG 28551	IV	CCUG
CCUG 29785	VI	CCUG
ATCC 49449	X	ATCC
ATCC 27956	not typed	ATCC
ATCC 7077	not typed	ATCC
ATCC 4768	not typed	ATCC
ATCC 12927	not typed	ATCC
ATCC 9925	not typed	ATCC

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Inclusivity Results
Bacterial Strain	Serotype	Vendor
ATCC 55194	not typed	ATCC
ATCC 55191	not typed	ATCC
CNCTC 6609	VII	CNCTC

f. Assay cut-off:
Not applicable.

2. Comparison studies:

Method comparison with predicate device: a.
Not applicable
b. Matrix comparison:
Not applicable
1. Clinical studies:
- a. Clinical Sensitivity:

Performance characteristics of the AmpliVue® GBS Assay were established during a prospective study conducted from July to November 2013. Nine hundred eleven specimens used for this study were collected from antepartum women between 35 to 37 weeks' gestation at four distinct geographical sites across the United States. ' The age range for these women was between 15 to 44 years old.

Nine hundred eleven specimens were tested by both the AmpliVue® GBS Assay and bacterial culture. Three specimens (0.3%) were invalid in the AmpliVue® GBS Assay when initially tested. Three specimens remained invalid upon repeat testing. We elected to calculate clinical performance based on the initial test result obtained for each specimen. Therefore, the data below is for the remaining nine hundred eight specimens.

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Combined Sites
	Bacterial Culture						95% CI
		POS	NEG				Total	Sensitivity
AmpliVue® GBS Assay	POS	196	52*	248	Specificity	92.7%	90.5%	94.3%
	NEG	1**	659	660
	Total	197	711	908

Fifty-two (52) discordant specimens (AmpliVue Positive/Bacterial Culture Negative) reported above were tested with an FDA-cleared molecular device for the detection of GBS. Thirty-nine (39) of these specimens were positive for GBS, and thirteen (13) were negative.

** The one (1) discordant specimen (AmpliVue Negative/Bacterial Culture Positive) reported above was tested with an FDA-cleared molecular device for the detection of GBS, and was negative.

b. Clinical specificity:
See Section 3a.
Other clinical supportive data (when a. and b. are not applicable); ﻥ
Not applicable
1. Clinical cut-off:
  Not applicable
1. Expected values:
  Clinical performance of the AmpliVue® GBS assay with Lim enrichment broth was established in specimens from antepartum women at four clinical sites during a clinical study performed in 2013. Nine hundred and eight specimens collected from antepartum women between 35 to 37 weeks' gestation at four distinct geographical sites across the United States, were tested. The age range for these women was between 15 to 44 years old. The percentage of positive cases as determined by the AmpliVue @ GBS assay during the study was 27.3% (248 of 908).

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Image /page/15/Picture/0 description: The image shows the text "DEPARTMENT OF HEALTH & HUMAN SERVICES" in bold, uppercase letters. The text is black and appears to be centered. The words are stacked on top of each other, with "DEPARTMENT OF HEALTH" on the top line and "& HUMAN SERVICES" on the bottom line.

Public Health Service

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

QUIDEL CORPORATION RONALD H. LOLLAR SENIOR DIRECTOR CLINICAL AND QUALITY AFFAIRS 2005 EAST STATE ST., SUITE 100 ATHENS OH 45701

December 20, 2013

Re: K133503

Trade/Device Name: AmpliVue® GBS Assay Regulation Number: 21 CFR 866.3740 Regulation Name: Streptococcus spp. serological reagents Regulatory Class: I Product Code: NJR Dated: November 13, 2013 Received: November 14, 2013

Dear Mr. Lollar:

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 Parts 801 and 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 electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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Page 2-Mr. Lollar

. ...

If you desire specific advice for vour device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Small Manufacturers, International and Consumer Assistance 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" (21CFR 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/ReportalProblem/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 Small Manufacturers, International and Consumer Assistance 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 yours.

Uwe Scherf -S for

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

Enclosure

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Indications for Use

510(k) Number (if known):

Device Name: AmpliVue GBS Assay

Indications For Use:

. .
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

The AmpliVue® GBS Assay is a qualitative in vitro diagnostic test for the rapid detection of Group B Streptococcus from vaginal/rectal swabs from antepartum women following 18 to 24 hours of incubation in an LIM enrichment broth culture. The AmpliVue® GBS Assay utilizes helicase-dependent amplification (HDA) of the Thiolase (atoB) gene sequence and a self-contained disposable amplification detection device that allows for manual evaluation of assay results. Results can be used as an aid in determining the colonization status of antepartum women.

The AmpliVue GBS Assay does not provide susceptibility results. Culture isolates are needed for performing susceptibility testing as recommended for penicillin-allergic women.

The AmpliVue GBS Assay is intended for use in hospital, reference or state laboratory settings. The device is not intended for point-of-care use.

Prescription Use _____________________________________________________________________________________________________________________________________________________________	AND/OR	Over-The-Counter Use
(Part 21 CFR 801 Subpart D)		(21 CFR 807 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH; Office of in Vitro Diagnostics and Radiological Health (OR)

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Page 1 of 1

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.