Diagnosing Bacterial Vaginosis/Vaginitis (BV) Using the Gynecologene Test Method

Bacterial vaginosis (BV, or vaginal bacteriosis) and vaginitis are the most common vaginal
infections of women during the child-bearing years, with BV alone accounting for 27-41% 1
prevalence in Americans. BV is characterized by replacement of the healthy
lactobacillus-dominant microbiome by a diverse polymicrobial overgrowth of multiple taxa of
commensal facultative anaerobes and other pathogenic bacteria. Molecular-based phylogenetic
analyses, including the newly-completed Human Microbiome Project and other next-generations
sequencing (NGS) studies, revealed the complexity of the vaginal microbiome in health and
disease. BV does not appear to be a single entity, but rather consists of multiple bacterial
community clusters. Clusters are dominated by Lactobacillus iners, Lactobacillus crispatus,
or anaerobic combinations with or without Lactobacillus spp. (species) in varying
proportions. In addition, multiple significant BV-associated pathogens were discovered that
had escaped notice with other detection methods such as cultures; previously overlooked were
Atopobium vaginae, Lactobacillus iners and other Lactobacillus spp., and multiple
Lachnospiraceae spp (Bacterial Vaginosis Associated 1, Bacterial Vaginosis Associated 2, and
Bacterial Vaginosis Associated 3; BVA1, BVA2 and BVA 3 respectively). This multisite study
proposes to evaluate the sensitivity and specificity of the Gynecologene Next-Generation
Sequencing test in women with symptoms of vaginitis/vaginosis. To the investigators'
knowledge, the Gynecologene is the first commercially-available comprehensive NGS-based
laboratory-developed test available to assist with diagnosis and clinical management of BV.

This multisite site study compares the diagnostic performance of the Gynecologene
Next-Generation Sequencing test with the clinical diagnosis of bacterial vaginosis/vaginitis
(Nugent score). The second objective is to compare the sensitivity and specificity of the
Gynecologene Score with the Nugent Score for diagnosis of bacterial vaginosis/vaginitis. The
third objective is to identify the prevalence and diagnostic thresholds of bacterial loads
with the Gynecologene test.

Women between 18-55 years of age will be eligible for this study when diagnosed with
bacterial vaginosis or vaginitis confirmed using the CDC (Centers for Disease Control)
diagnostic gold standard of Nugent score; Amsel criteria will also be recorded. Other
inclusion as well as exclusion criteria exist. A minimum of 5 geographically different sites
will be enrolled in the study. Co-investigators will follow the protocol to enroll
participants and collect 2 vaginal swabs and 1 slide smear. The patient has no financial
gain nor responsibility within this protocol.

Overall, the study will have 90% power at a significance level of 0.05 to detect ± 5%
positive percent agreement of the study device with the predicate device. To achieve this
level of power, 100 total samples will be assayed per group to account for withdrawals and
treatment failures. Samples that contain more than one targeted organism can be counted
twice as the analyses for each organism are independent of each other. The provision of
treatment, including type, dose, and duration, is solely at the discretion of the clinician
according to his/her current routine procedures to reflect typical contemporary practice.

AIBiotech will serve as a central medical diagnostic laboratory for this trial. Results of
all analytical tests will be provided to the Principal Investigator.

Data for analysis will be blinded for patient identification and other PHI (protected health
information) by the study administrator. Data will be analyzed and reported.

Inclusion Criteria:

- Women ≥ 18 years of age and any race (African-American, Hispanic, Asian/Pacific
Islander, native American, white).

- Women during reproductive years prior to menopause and in no case over age 55 years.

- Signed Informed Consent document obtained prior to the initiation of screening
procedures.

- Signs and symptoms of vaginitis/vaginosis, including vaginal discharge, pruritis,
irritation, and/or dyspareunia.

- Diagnosis of bacterial vaginosis (study group) according to Nugent score (see below);
and/or diagnosis of vulvovaginal candidiasis; and/or diagnosis of Trichomonas
vaginalis.

- Clinical Diagnosis: Nugent Score: The presence of abnormal vaginal flora at initial
diagnosis confirmed by gram stain, with all subjects with BV (bacterial vaginosis or
bacterial vaginitis) having Nugent score > 7.

- Inclusion Criteria of Control Group: same as Study Group, but Nugent Score < 7.

- No active major medical or psychological problems that could be complicated by study
participation.

Exclusion Criteria:

- Treatment with any investigational prescription product within 28 days of study
screening.

- Post-menopausal or greater than 55 years of age.

- Prior history of hysterectomy or vaginal surgery.

- Prior or currently active autoimmune disease requiring management with systemic
immunosuppression for any reason. Such conditions include inflammatory bowel disease,
systemic vasculitis, scleroderma, psoriasis, multiple sclerosis, hemolytic anemia,
immune-related thrombocytopenia, rheumatoid arthritis, systemic lupus erythematosus,
Sjögren's syndrome, sarcoidosis, or other rheumatological disease.

- Any infection requiring parenteral/ enteral antibiotic therapy or causing fever (body
temperature > 100.5°F or 38.1°C) within 4 weeks prior to study screening.

- Any medical intervention or other condition which, in the opinion of the
Physician-Investigator could compromise adherence with study requirements or
otherwise compromise study subject safety and the study's objectives.