BVBLUE™ is an enzyme activity test for use in the detection of vaginal fluid specimens for sialidase activity, an enzyme produced by bacterial pathogens such as Gardnerella vaginalis, Bacteroides spp., Prevotella spp., and Mobiluncus spp.

BVBLUE™ is indicated for use in women suspected of having Bacterial Vaginosis (BV) infection, e.g., women with vaginal discharge typical of BV and/or women with previous history of BV, as an aid in the diagnosis of BV infection. BVBLUE™ test results should be considered in conjunction with other clinical and patient information (see Limitations of the Procedure).

For In Vitro Diagnostic Use Only. BVBLUE™ is indicated for professional use only and may be used at the point of care and/or in physician's offices. It is not intended for home use.

Device Description

The BVBLUE™ Test contains a chromogenic substrate of sialidase enzyme, IBX-4041. The chromogenic substrate is provided in each of the BVBLUE™ Testing Vessels as a solution containing 0.25 mg IBX-4041 in 0.5 mL aqueous potassium acetate solution (49.0 mg/mL; 0.5 M, pH 5.5-6.0). When the solution containing the solubilized chromogenic substrate is exposed to sialidase enzyme, the substrate is hydrolyzed to yield sialic acid and IBX-4050. Upon the addition of one of more drops of the BVBLUE™ Developer Solution [an aqueous sodium hydroxide solution (40 mg/mL; 1.0 M; pH>11.0)], IBX-4050 exhibits an intense blue color. In the absence of sialidase enzyme, the chromogenic substrate is not hydrolyzed and exhibits a vellow color upon the addition of one or more drops of the BVBLUE'M Developer Solution.

Materials Provided:
10 BVBLUE™ Units per Package:

Ten (10) BVBLUE™ Testing Vessels each containing 0.25 mg IBX-4041 . component in 0.5 mL of an aqueous potassium acetate buffer solution (49.0 mg/mL; 0.5 M; pH 5.5-6.0).
One (1) BVBLUE™ Developer Solution Tube containing 0.5 mL of an . aqueous sodium hydroxide solution (40.0 mg/mL; 1.0 M; pH>11.0).
Instructions for Use .

AI/ML Overview

Here's a detailed breakdown of the acceptance criteria and study information for the BVBLUE™ device, based on the provided text:

BVBLUE™ Device: Acceptance Criteria and Study Details

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state "acceptance criteria" in a separate section with specific numerical targets prior to presenting results. Instead, performance metrics are reported, and the conclusion states that these demonstrate substantial equivalence. Based on the provided performance numbers, the implied acceptance criteria would be those achieved or exceeded in the studies.

Performance Metric	Implied Acceptance Criteria (Achieved)	Reported Device Performance	Study Context (e.g., population size)
Non-Clinical Testing:
Analytical Sensitivity (BVBLUE™ positive for ≥0.25 µg sialidase)	100% Identification	100% (69/69)	207 replicates (69 positive, 138 negative)
Analytical Specificity (BVBLUE™ negative for <0.25 µg sialidase)	100% Identification	100% (138/138)	207 replicates (69 positive, 138 negative)
Inter-operator Reproducibility (Positive, ≥0.30 µg sialidase)	100% Identification	100% (18/18)	90 replicates (18 positive, 72 negative)
Inter-operator Reproducibility (Negative, <0.30 µg sialidase)	≥96% Identification	96% (69/72)	90 replicates (18 positive, 72 negative)
Real-time Stability	≥241 days (and continuing)	241 days (and continuing)	Ambient temperature
Accelerated Stability	≥41 days	41 days	56 °C
Clinical Study 1 (vs. Amsel Criteria, initial findings):
BVBLUE™ Sensitivity	≥85%	85% (73/86)	86 samples positive by Amsel Criteria
BVBLUE™ Specificity	≥90%	90% (375/416)	416 samples negative by Amsel Criteria
Clinical Reproducibility	100% Correlation	100%	50 vaginal fluid samples from 25 women
Clinical Study 2 (vs. Amsel Criteria, reconciled):
BVBLUE™ Sensitivity	≥97%	97% (30/31)	31 samples positive by Amsel Criteria
BVBLUE™ Specificity	≥99%	99% (86/87)	87 samples negative by Amsel Criteria
Clinical Study 2 (vs. Gram's Stain, reconciled):
BVBLUE™ Sensitivity	≥97%	97% (30/31)	31 samples positive by Gram's stain
BVBLUE™ Specificity	≥99%	99% (86/87)	87 samples negative by Gram's stain
Clinical Study 2 (vs. Amsel OR Gram's Stain):
BVBLUE™ Sensitivity	≥76%	76% (26/34)	34 samples positive by Amsel OR Gram's stain
BVBLUE™ Specificity	≥98%	98% (82/84)	84 samples negative by Amsel OR Gram's stain
Clinical Study 2 (vs. Semiquantitative Morphotpyes):
BVBLUE™ Sensitivity	≥70%	70% (30/43)	43 samples positive by Morphotpyes
BVBLUE™ Specificity	≥99%	99% (74/75)	75 samples negative by Morphotpyes
Clinical Study 2 (vs. Culture of Microorganisms):
BVBLUE™ Sensitivity	≥77%	77% (10/13)	13 samples positive by Culture
BVBLUE™ Specificity	≥83%	83% (35/42)	42 samples negative by Culture
Clinical Study 2 (vs. Clinical Diagnosis of BV):
BVBLUE™ Sensitivity (BV only)	≥96%	96% (48/50)	50 samples with BV diagnosis
BVBLUE™ Sensitivity (BV + Candidiasis)	≥80%	80% (4/5)	5 samples with BV + Candidiasis
BVBLUE™ Sensitivity (BV + Trichomoniasis)	100%	100% (2/2)	2 samples with BV + Trichomoniasis
BVBLUE™ Specificity (Gonorrhea)	100%	100% (1/1)	1 sample with Gonorrhea
BVBLUE™ Specificity (Herpes)	100%	100% (1/1)	1 sample with Herpes
BVBLUE™ Specificity (Trichomoniasis)	100%	100% (4/4)	4 samples with Trichomoniasis
BVBLUE™ Specificity (Candidiasis)	100%	100% (41/41)	41 samples with Candidiasis
BVBLUE™ Specificity (Healthy Controls)	≥94%	94% (142/151)	151 healthy control samples
Sialidase enzyme activity (BV patients)	95% CI significantly higher than cutoff	95% CI (8.12-16.56 U)	27 BV patients, mean 12.34 U
Sialidase enzyme activity (Non-BV patients: Candidiasis, Trichomoniasis, Healthy)	95% CI significantly lower than cutoff	Candidiasis (2.57-4.83 U), Trichomoniasis (0.59-3.39 U), Healthy (2.23-3.17 U)	59 healthy, 16 Candidiasis, 3 Trichomoniasis

2. Sample Size and Data Provenance

Non-Clinical Test Set:
- Analytical Samples: 207 replicates (69 positive, 138 negative) of coded sialidase solutions.
- Inter-operator Reproducibility: 90 replicates (18 positive, 72 negative) of coded sialidase solutions.
- Provenance: Analytical laboratory setting. Country of origin not specified, but implied domestic (USA) where the company is located. These are retrospective as they are prepared solutions, not patient samples.
Clinical Study 1 Test Set:
- Total Sample Size: 502 women.
- Test Set Size (BVBLUE vs. Amsel): 502 samples.
- Test Set Size (Reproducibility): 50 vaginal fluid samples from 25 women.
- Provenance: Conducted at 5 separate clinical sites. Country of origin not specified, but implied domestic (USA). This was a prospective study where samples were collected and tested.
Clinical Study 2 Test Set:
- Total Sample Size: 118 women.
- Test Set Size (BVBLUE vs. Amsel or Gram's Stain): 118 samples for Amsel, Gram's Stain, Morphotpyes, and quantitative sialidase.
- Test Set Size (BVBLUE vs. Culture): 55 samples.
- Test Set Size (Clinical Diagnosis & Sialidase Activity): 255 clinical samples (clinical diagnosis comparison), 118 clinical samples (quantitative sialidase activity).
- Provenance: Conducted at 2 separate clinical sites. Country of origin not specified, but implied domestic (USA). This was a prospective study.

3. Number of Experts and Qualifications for Ground Truth

The document does not explicitly state the number or qualifications of "experts" used to establish ground truth in the traditional sense of independent readers reviewing cases. Instead, the ground truth for clinical studies was established using recognized diagnostic methods:

Amsel Criteria: This is described as the "Gold Standard Method of clinically diagnosing bacterial vaginosis as recommended by the CDC." It involves a set of clinical signs and symptoms evaluated by a clinician, along with specific laboratory findings (e.g., pH, amine odor). The personnel involved in applying Amsel Criteria were "site personnel" (18 in Study 1, 4 in Study 2) and "company employees" (2 in Study 1, 1 in Study 2). Their specific qualifications (e.g., experience as physicians, nurses, lab technicians) are not detailed, but their role in applying a "Gold Standard" implies medical or laboratory training.
Gram's Stain: Described as "a second Gold Standard Method of clinically diagnosing bacterial vaginosis." The personnel performing and interpreting Gram's stain are not specifically detailed for their qualifications, but would typically be trained laboratory technologists or physicians.
Semi-Quantitative Identification of Morphotpyes, Genital Culture, Anaerobic Culture: These are laboratory-based diagnostic methods requiring trained laboratory personnel.

4. Adjudication Method for the Test Set

The document mentions "reconciled comparison" in Clinical Study 2 for BVBLUE™ vs. Amsel Criteria and BVBLUE™ vs. Gram's Stain. This implies some form of adjudication for discordant cases between the BVBLUE™ test and the reference standard. However, the specific method of adjudication (e.g., a third expert review, consensus among original reviewers, or specific protocol for resolving discrepancies) is not detailed. For Clinical Study 1, it is explicitly stated that "The first clinical study did not provide for the evaluation of discordant samples via a second reference method..." indicating no explicit adjudication for discordant results in that study beyond the initial comparison.

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

No Multi-Reader Multi-Case (MRMC) comparative effectiveness study was done to evaluate the effect size of how much human readers improve with AI vs without AI assistance. The BVBLUE™ is a diagnostic test kit to be used by trained personnel, not an AI assistance tool for human interpretation. The "inter-operator reproducibility studies" evaluated the consistency of the test kit results when performed by different individuals, not a human reader's diagnostic performance with or without AI.

6. Standalone (Algorithm Only) Performance

Yes, the BVBLUE™ device performance as a standalone test kit for detecting sialidase activity (and thus aiding in BV diagnosis) was evaluated. The performance metrics presented for sensitivity and specificity against the ground truth (Amsel Criteria, Gram's Stain, etc.) represent the standalone performance of the device without explicit human-in-the-loop diagnostic assistance for interpretation beyond performing the test itself and reading its color change.

7. Type of Ground Truth Used

The ground truth for the clinical studies was primarily established by:

Expert Consensus/Clinical Standard: Amsel Criteria, which combines clinical observation and simple lab tests (pH, amines). This is explicitly called the "Gold Standard" as recommended by the CDC.
Laboratory Standard: Gram's stain, also described as a "Gold Standard" for bacterial vaginosis diagnosis, involving microscopic evaluation by trained personnel.
Other Laboratory Methods: Semi-quantitative identification of morphotypes, genital cultures, and anaerobic cultures were also used as reference methods in Clinical Study 2.
Clinical Diagnosis: Comparison against a broader clinical diagnosis of various vaginal conditions (bacterial vaginosis, candidiasis, trichomoniasis, gonorrhea, herpes).
Quantitative Sialidase Enzyme Activity: Used to establish the quantitative levels of the target enzyme in different clinical conditions and compare them to the device's cutoff.

8. Sample Size for the Training Set

The document does not explicitly state details about a "training set" for the device in the context of machine learning or algorithm development. The BVBLUE™ is described as an enzyme activity test with a chromogenic substrate to identify sialidase. It is not an algorithm that would typically undergo a separate "training" phase with a dataset. The non-clinical studies (analytical sensitivity/specificity, inter-operator reproducibility) could be considered part of the initial validation and refinement.

9. How Ground Truth for the Training Set Was Established

As noted above, there is no explicit "training set" in the context of an algorithm. For the non-clinical studies concerning analytical performance and reproducibility, the ground truth was established by preparation of coded sialidase solutions with known concentrations (e.g., ≥0.25 µg or <0.25 µg of sialidase enzyme).

Summary

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MAY 1 5 2000

EXTUBIT # I Page 1 of 8

510(K) SUMMARY

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is:__K993732/S1

1. Submitter's Identification:

Gryphus Diagnostics, L.L.C. 2800 Milan Court, Suite 235 Birmingham, AL 35211 Contact Person: Stephen C. Johnson, Ph.D.

Data Summary Prepared: October 20, 1999

Name of the Device: 2.

BVBLUETM

3. Predicate Device Information:

1. Affirm/VP Identifiaction Test for Gardnerella vaginalis and Trichomonas vaginalis, K# 923133, Microprobe Inc.
1. Indicard® Test, K# 964015, Litmus Concepts, Inc.
1. FemExam® TestCard™ pH and Amines Tests, K# 962718, Litmus Concepts, Inc.

Device Description: র্বা

Materials Provided:

10 BVBLUE™ Units per Package:

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Ten (10) BVBLUE™ Testing Vessels each containing 0.25 mg IBX-4041 . component in 0.5 mL of an aqueous potassium acetate buffer solution (49.0 mg/mL; 0.5 M; pH 5.5-6.0).
One (1) BVBLUE™ Developer Solution Tube containing 0.5 mL of an . aqueous sodium hydroxide solution (40.0 mg/mL; 1.0 M; pH>11.0).
Instructions for Use .

ર. Intended Use:

BVBLUE™ is indicated for use in wornen suspected of having Bacterial Vaginosis (BV) infection, e.g., women with vaginal discharge typical of BV and/or women with previous history of BV, as an aid in the diagnosis of BV infection. BVBLUE™ test results should be considered in conjunction with other clinical and patient information (see Limitations of the Procedure).

For In Vitro Diagnostic Use Only. BVBLUE™ is indicated for professional use only and may be used at the point of care and/or in physician's offices. lt is not intended for home use.

6. Comparisons to Predicate Devices:

Affirm/VP (manufactured by Becton Dickinson) evaluates patient vaginal fluid samples for the presence of Gardnerella vaginalis nucleic acid. Gardnerella vaginalis is a microorganism causative of bacterial vaginosis. Using this test, a patient vaginal fluid sample is obtained with a sterile cotton swab. A portion of the sample is transferred to a tube containing a lysis solution, thoroughly mixed, incubated at 85 °C for 10 minutes, and treated with a buffer solution. A portion of the resulting mixture is transferred to a Processor and then reacted for a period of 30 minutes to provide an identification of Gardnerella vaginalis nucleic acid. A positive indication for Gardnerella vaginalis nucleic acid is indicative of bacterial vaginosis in patients presenting bacterial vaginosis-like symptoms.

Indicard™ test (manufactured by Litmus Concepts, Inc.) evaluates patient vaginal fluid specimens for the presence of Gardnerella vaginalis proline imino peptidase enzyme activity. Gardnerella vaginalis is a microorganism

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causative of bacterial vaginosis. Using this test, a patient vaginal fluid sample is obtained with a sterile cotton swab. A portion of the sample is then transferred to the Indicard™ test to evaluate for the presence of Gardnerella vaginalis. A positive indication for Gardnerella vaginalis oroline imino peotidase enzyme activity is indicative of bacterial vaginosis in patients presenting bacterial vaginosis-like symptoms.

FemExam® TestCard™ (manufactured by Litmus Concepts, Inc.) evaluates patient vaginal fluid samples for elevated pH (i.e., pH >4.7) and for the presence of volatile amines in patient vaginal fluid samples. The pH and amine tests are two components of the Amsel Criteria, the Gold Standard method of clinically diagnosing patient vaginal fluid samples for bacterial vaginosis. Using this test, a patient vaginal fluid sample is obtained with a sterile cotton swab. A portion of the sample is first transferred to a section of the FemExam® TestCard™ to evaluate for elevated pH. A second portion of the sample is transferred to the card to evaluate for the presence of volatile amines. A positive indication in both the pH and amines tests is indicative of bacterial vaginosis in patients presenting bacterial vaginosis-like symptoms.

BVBLUE TM (manufactured by Gryphus Diagnostics, L.L.C.) evaluates patient vaginal fluid samples for sialidase activity, an enzyme produced by bacterial pathogens associated with Bacterial Vaginosis (BV) such as Gardnerella vaginalis, Bacteroides spp., Prevotella spp., and Mobiluncus spp. Using this test, a patient vaginal fluid sample is obtained with a sterile cotton swab. A portion of the sample is then transferred to the BVBLUE™ test and incubated for 10 minutes at 37 ℃ to provide identification of sialidase enzyme activity. A positive indication for sialdiase enzyme activity is indicative of bacterial vaginosis in patients presenting bacterial vaginosis-like symptoms.

7. Discussion of Non-Clinical Tests Performed for Determination of Substantial Equilivance are as follows:

BVBLUE™ was evaluated using analytical samples of sialidase enzyme (Sigma Chemical Co.). The BVBLUE™ Test produces a positive result when the vaginal fluid contains ≥0.25 µg sialidase enzyme [i.e., ≥ 7.64 U (units of enzyme activity)]. This performance was defined in laboratory studies employing coded sialidase solutions containing varying levels of sialidase activity. A total of 207 replicates were evaluated.

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Of the 207 replicates, a total of 69/207 contained sialidase enzyme levels ≥0.25 ug (33%): and 138/207 contained sialidase enzyme levels <0.25 µg (67%).

BVBLUE™ properly identified 69/69 (100%) of confirmed positives for sialidase enzyme levels ≥0.25 µg from analytical samples. BVBLUE™ properly identified 138/138 (100%) of confirmed negatives for sialidase enzyme levels <0.25 ug from analytical samples.

Inter-operator reproducibility studies were conducted with BVBLUE 1M by three individuals in a blinded study employing coded sialidase solutions of varying levels of sialidase activity. A total of 30 replicates were performed by each individual, for a total of 90 replicates.

Of the 90 replicates, a total of 18/90 contained sialidase enzyme levels of 20.30 µg (20%); and 72/90 contained sialidase enzyme levels of <0.30 µg (80%).

BVBLUE™ properly identified 18/18 (100%) of confirmed positives for sialidase enzyme levels ≥0.30 µg from analytical samples. BVBLUE™ properly identified 69/72 (96%) of confirmed negatives for sialidase enzyme levels <0.30 µg from analytical samples.

Real-time and accelerated stability studies were conducted with BVBLUE™. Accelerated studies conducted at 56 °C established a stability of 41 days at 56 °C. Real-time studies conducted at ambient temperature established a stability of 241 days and continuing. These real-time stability studies are continuing.

8. Discussion of Clinical Tests Performed:

BVBLUE™ was evaluated in two separate clinical studies.

The first clinical study was conducted by 18 site personnel and 2 company employees at 5 separate clinical sites. A total of 502 women were evaluated. The objective of this study was to define correlations between a diagnosis of bacterial vaginosis based on Amsel Criteria, the Gold Standard Method of clinically diagnosing bacterial vaginosis as recommended by the CDC, and BVBLUE™. A second objective was to determine the reproducibility of BVBLUE™ in patients with and without bacterial vaginosis. Two vaginal fluid samples were obtained from all women by contacting a sterile cotton swab with the vaginal wall (lower one-third). One swab sample was used in an evaluation for bacterial vaginosis based on Amsel Criteria and the second was used in an evaluation of BVBLUE™. A total of three vaginal fluid samples were

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obtained from a subset of women attending one clinical site. The third
swab sample was used to perform an independent BVBLUE™ Test. The results of the two tests were compared. Women were excluded from the study if they had used an antibiotic or antifungal agent within three days of testing. One production lot of the BVBLUE™ Test Kit was used in the clinical study.

Of the 502 clinical samples evaluated, a total of 86/502 (17%) exhibited bacterial vaginosis based on Amsel Criteria. Of these 86 samples, 73 (85%) exhibited a positive BVBLUE™ test result. Of the 502 clinical samples evaluated, a total of 416/502 (83%) did not exhibit bacterial vaginosis based on Amsel Criteria. Of these 416 samples, 375 (90%) exhibited a negative BVBLUE™ test result.

The first clinical study did not provide for the evaluation of discordant samples via a second reference method of diagnosing for bacterial vaginosis. This issue, among others, was addressed in a second clinical study.

There was a 100% correlation of results during the reproducibility testing of BVBLUE™. The reproducibility of the BVBLUE Test was conducted on a total of 50 vaginal fluid samples obtained from a total of 25 women.

The second clinical study was conducted by 4 site personnel and 1 company employee at 2 separate clinical sites. A total of 118 women were evaluated. The objective of this study was to (1) define correlations between a diagnosis of bacterial vaginosis based on Amsel Criteria, the Gold Standard Method of clinically diagnosing bacterial vaginosis as recommended by the CDC, and BVBLUE™, (2) define correlations between a diagnosis of bacterial vaginosis based on Gram's stain, a second Gold Standard Method of clinically diagnosing bacterial vaginosis, and BVBLUE™: (3) define correlations between a diagnosis of bacterial vaginosis based on Amsel Criteria OR Gram's Stain and BVBLUE™: (4) define correlations between the semi-quantitative method of identifying Gardnerella vaginalis morphotypes, Prevotella spp. morphotypes, Bacteroides spp. morphotypes, and Mobiluncus spp. morphotypes and BVBLUE™; (5) define correlation between a genital culture for Gardnerella vaginalis, and anaerobic culture for microorganisms including Bacteroides spp. and Prevotella spp., and BVBLUETM; (6) define the quantitative sialidase enzyme activities in vaginal fluid specimens of women with bacterial vaginosis, candidiasis, trichomoniasis, and healthy controls; and (10) define the correlations between interfering substances including blood, semen, menses, contraceptive method, and vaginal signs and symptoms, and BVBLUE™ Three vaginal fluid samples were obtained from all women by contacting a sterile cotton swab with the vaginal wall (lower one-third). One swab sample was used in an evaluation of

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bacterial vaginosis based on Amsel Criteria; a second for an evaluation of bacterial vaginosis based on Gram's stain and the semi-quantitative identification of Gram's stain morphotypes; and a third for an evaluation of BVBLUE 100 and quantitative sialidase enzyme activity. A total of five vaginal fluid samples were obtained from a subset of women attending two clinical sites. The fourth swab sample was used in a genital culture. The fifth swab sample was used in an anaerobic culture. Women were excluded from the study if they had used an antibiotic or antifungal agent within three days of testing. Two production lots of the BVBLUE™ Test Kit was used in the clinical study.

In the reconciled comparison of BVBLUE™ vs. Amsel Criteria in all patients, of the 118 clinical samples evaluated, a total of 31/118 (26%) exhibited bacterial vaginosis. Of these 31 samples, 30 (97%) exhibited a positive BVBLUE™ test result. Of the 118 clinical samples evaluated, a total of 87/118 (74%) did not exhibit bacterial vaginosis. Of these 87 samples, 86 (99%) exhibited a negative BVBLUE™ test result.

In the reconciled comparison of BVBLUE™ vs. Gram's stain in all patients. of the 118 clinical samples evaluated, a total of 31/118 (26%) exhibited bacterial vaginosis. Of these 31 samples, 30 (97%) exhibited a positive BVBLUE™ test result. Of the 118 clinical samples evaluated, a total of 87/118 (74%) did not exhibit bacterial vaginosis. Of these 87 samples, 86 (99%) exhibited a negative BVBLUE™ test result.

In the comparison of BVBLUE™ vs. Amsel Criteria OR Gram's Stain in all patients, of the 118 clinical samples evaluated, a total of 34/118 (29%) exhibited bacterial vaginosis. Of these 34 samples, 26 (76%) exhibited a positive BVBLUE™ test result. Of the 118 clinical samples evaluated, a total of 84/118 (71%) did not exhibit bacterial vaginosis. Of these 84 samples, 82 (98%) exhibited a negative BVBLUE™ test result.

In the comparison of BVBLUE™ vs. Semi-Quantitative Identification of Morphotpyes Associates with Bacterial Vaginosis in all patients, of the 118 clinical samples evaluated, a total of 43/118 (36%) exhibited bacterial vaginosis. Of these 43 samples, 30 (70%) exhibited a positive BVBLUE™ test result. Of the 118 clinical samples evaluated, a total of 75/118 (64%) did not exhibit bacterial vaginosis. Of these 75 samples, 74 (99%) exhibited a negative BVBLUE™ test result.

In the comparison of BVBLUE™ vs. Clinically Significant Culture of Microorganisms Associated with Bacterial Vaginosis in all patients, of the 55 clinical samples evaluated, a total of 13/55 (24%) exhibited bacterial vaginosis. Of these 13 samples, 10 (77%) exhibited a positive BVBLUE™ test result. Of the 55 clinical samples evaluated, a total of 42/55 (76%) did

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not exhibit bacterial vaginosis. Of these 42 samples, 35 (83%) exhibited a negative BVBLUE™ test result.

In the comparison of BVBLUE™ vs. clinical diagnosis of bacterial vaginosis, Candidiasis, Trichomoniasis, herpes, gonorthea, and in healthy controls, of 255 clinical samples evaluated, a total of 50/255 (20%) exhibited bacterial vaginosis, 5/255 (2%) exhibited bacterial vaginosis and Candidiasis, and 2/255 (0.8%) exhibited bacterial vaginosis and Trichomoniasis. Of these samples, a total of 48/50 (96%) of those exhibiting bacterial vaginosis, 4/5 (80%) of those exhibiting bacterial vacinosis and Candidiasis, and 2/2 (100%) of those exhibiting bacterial vaginosis and Trichomoniasis exhibited a positive BVBLUE™ test result. Of the 255 clinical samples evaluated, a total of 1/255 (0,4%) exhibited gonorrhea. 1/255 (0.4%) exhibited herpes, 4/255 (1.6%) exhibited Trichomoniasis, 41/255 (16%) exhibited Candidiasis, and 151/255 (59%) were healthy controls. Of these samoles, 1/1 (100%) of those exhibiting gonorthea, 1/1 (100%) of those exhibiting herpes, 4/4 (100%) of those exhibiting Trichomoniasis. 41/41 (100%) of those exhibiting Candidiasis, and 142/151 (94%) of healthy controls exhibited a negative BVBLUE™ test result.

In the comparison of BVBLUE™ vs. the quantitative assessment of sialidase enzyme activity in patients with bacterial vaginosis. Candidiasis. Trichomoniasis, and in healthy controls, of 118 clinical samples evaluated. 27/118 (23%) exhibited bacterial vaginosis. The mean sialidase enzyme activity in these samples was 12.34 U. Of the 118 dinical samples, 59/118 (50%) were healthy controls. The mean sialidase enzyme activity in these samples was 2.70 U. Of the 118 clinical samples, 16/118 (14%) exhibited Candidiasis. The mean sialidase enzyme activity in these samples was 3.70 U. Of the 118 clinical samples, 3/118 (3%) exhibited Trichomoniasis. The mean sialidase enzyme activity in these samples was 1.99 U.

The cutoff precision of BVBLUE™, as determined using analytical samples of sialidase enzyme, was found to be 7.64 U. From the clinical studies, the 95% CI for patients with bacterial vaginosis (8.12-16.56 U) is significantly higher than the cutoff of BVBLUE™ The 95% CIs for patients with Candidiasis (2.57-4.83 U), Trichomoniasis (0.59-3.39 U), or in healthy controls (2.23-3.17 U) are significantly lower than the cutoff of BVBLUETM

The data from this clinical study demonstrated that blood, semen, menses, contraceptive methods, vulvo vaginal signs and symptoms, and the presence of other morphotypes and microorganisms did not adversely effect the performance of BVBLUE 10

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EXHIBIT # 1 Page 8 of 8

9. Conclusions:

The BVBLUE™ device has similar intended uses and similar technological characteristics as the predicate devices. Moreover, bench testing contained in this submission and clinical testing supplied demonstrate that contaiffed in this cubeir sechnological characteristics do not raise any new any different of safety or effectiveness. Thus, the BVBLUE™ device is qubstantially equivalent to the predicate devices in the identification of bacterial vaginosis in patient vaginal fluid samples.

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Image /page/8/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is a stylized symbol of three human profiles facing right, stacked on top of each other, representing health and human services.

MAY 1 5 2000

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Gryphus Diagnostics, L.L.C. c/o Mr. Thomas M. Tsakeris Devices and Diagnostics Consulting Group 16809 Briardale Road Rockville, Maryland 20855

Re: K993732

Trade Name: BVBlue™ Regulatory Class: I Reserved Product Code: MXB Dated: March 7, 2000 Received: March 7, 2000

Dear Mr. Tsakeris:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). 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.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic QS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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Page 2

This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device. please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll-free number (800) 638-2041 or (301) 443-6597 or at its internet address "http://www.fda.gov/cdrl/dsma/dsmamain.html".

Sincerely yours,

Steven Butman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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EXHIBIT B Page 1 of 1

510(k) Number (if known): K993732

Device Name: BV BLUE™

Indications for Use:

BV BLUEm is an enzyme activity test for use in the detection of vaginal fluid specimens for sialidase activity, an enzyme produced by bacterial pathogens such as Gardnerella vaginalis, Bacteroides spp., Prevotella spp., and Mobiluncus spp.

BV BLUE™ is indicated for use in women suspected of having Bacterial Vaginosis (BV) infection, e.g., women with vaginal discharge typical of BV and/or women with previous history of BV, as an aid in the diagnosis of BV infection. BV BLUE™ test results should be considered in conjunction with other clinical and patient information (see Limitations of the Procedure)

For In Vitro Diagnostic Use Only. BV9LUE™ is indicated for professional use only and may be used at the point of care and/or in physician's offices. It is not intended for home-use.

Cooper
(Division Sign-Off)
Division of Clinical Laboratory Devices
510(k) Number K993732

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Concurrence of CDRH, Office of Device Evaluation (ODE)

い OR Over-The-Counter Use Prescription Use Format 1-2-96) (Per 21 CFR 801.109)

Regulation Number and Section

§ 866.2660 Microorganism differentiation and identification device.

(a)
Identification. A microorganism differentiation and identification device is a device intended for medical purposes that consists of one or more components, such as differential culture media, biochemical reagents, and paper discs or paper strips impregnated with test reagents, that are usually contained in individual compartments and used to differentiate and identify selected microorganisms. The device aids in the diagnosis of 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.