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

Prospective Multicenter Cross Sectional Study of the American International Biotechnology (AIBiotech) Gynecologene Test for the Evaluation of Bacterial Vaginosis/Vaginitis in Symptomatic Women

This study is designed as a prospective evaluation of the diagnostic performance of the multiplex nucleic acid-based genetic test (Gynecologene Next-Generation Sequencing test) to identify known significant causative organisms in bacterial vaginosis/vaginitis and other major pathogens and normal commensals in symptomatic women during the reproductive years. Vaginal fluid samples will be split and tested with the comparator methods and Nugent score, with the results evaluated according to sensitivity, specificity, positive predictive value, and negative predictive value. Laboratorians performing each test will be blinded to the clinical history and any prior test results to minimize ascertainment bias.

Each subject will be followed routinely for up to 6 weeks after initial visit for vaginosis/vaginitis by telephone or office visit at the discretion of the treating physician.

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.