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Female Ecologixtm Vaginal Health & Microbiome Profile Phylo Bioscience Laboratory

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Female Ecologixtm Vaginal Health & Microbiome Profile Phylo Bioscience Laboratory Female EcologiXTM Vaginal Health & Microbiome Profile Phylo Bioscience Laboratory INTERPRETIVE GUIDE DISCLAIMER: THIS INFORMATION IS PROVIDED FOR THE USE OF PHYSICIANS AND OTHER LICENSED HEALTH CARE PRACTITIONERS ONLY. THIS INFORMATION IS NOT FOR USE BY CONSUMERS. THE INFORMATION AND OR PRODUCTS ARE NOT INTENDED FOR USE BY CONSUMERS OR PHYSICIANS AS A MEANS TO CURE, TREAT, PREVENT, DIAGNOSE OR MITIGATE ANY DISEASE OR OTHER MEDICAL CONDITION. THE INFORMATION CONTAINED IN THIS DOCUMENT IS IN NO WAY TO BE TAKEN AS PRESCRIPTIVE NOR TO REPLACE THE PHYSICIANS DUTY OF CARE AND PERSONALISED CARE PRACTICES. INTERPRETIVE GUIDE Female EcologiX™ INTRODUCTION Due to recent advancements in culture-independent molecular techniques, it is now possible to measure the composition of the human microbiota. One key microbiota site is the vaginal tract, which hosts over 200 species of microorganisms1. The vaginal microbiota is a dynamic and complex ecosystem comprised of fluctuating populations that modulate host immune responses and maintain homeostasis. Disruption of microbiota composition and functions, termed dysbiosis, has been linked to a multitude of disorders including bacterial vaginosis (BV), premature delivery in pregnant women, infertility, miscarriages and increased risk of sexually transmitted infections (STIs). Analysis and monitoring of the vaginal microbiota is therefore extremely important for health and can be used to inform treatment plans and lifestyle factors. To meet this need, Phylobioscience have developed Female EcologiXTM Vaginal Health and Microbiome Profile, a ground breaking tool for analysis of vaginal microbiota composition and host immune responses. Using innovative microbial culture- independent technologies, including quantitative real-time PCR (qRT-PCR) and enzyme-linked immunosorbent assays (ELISA), the profile provides an accurate, reliable and quantifiable measurement of microbiota abundance and markers for host immune responses. For microbiota composition analysis, the technology detects: • Abundance of four key Lactobacillus species (markers for vaginal health) • Abundance of strictly anaerobic bacteria (associated with dysbiosis) • Abundance of enteric bacteria including Escherichia coli and Staphylococcus aureus (associated with aerobic vaginitis) • Presence of STI pathogen Chlamydia trachomatis • Presence of STI viral pathogens herpes simplex virus type 1 (HSV-1) and HSV-2 (genital herpes) • Abundance of BV-associated bacteria including Gardenella, Prevotella and Megasphaera species For host biomarker analysis, the technology detects: • I L-1 β (proinflammatory cytokine) Dependent on microbiota composition and host biomarkers, the profile will report three different states: • Healthy vaginal microbiota (homeostasis) • Vaginal dysbiosis (microbiota profile and/or host marker profile) • STI pathogen detected (Chlamydia, HSV-1, HSV-2) Invivo Healthcare, The New Warehouse, Libby’s Drive, Stroud, GL5 1RN page 1 Please call us on 0333 241 2997, or visit us at invivohealthcare.com INTERPRETIVE GUIDE GLOSSARY Term Description References Commensal Microorganism (i.e. bacteria, fungi) that lives in symbiosis with host when residing within 2 its specific environment Pathogen Microorganism (e.g. bacteria, fungi, virus) that may cause disease 3 Pathobiont Potential pathogen that lives in symbiosis under normal conditions 4 Homeostasis Ability to maintain internal stability in an organism despite environmental changes Dysbiosis Imbalance or disturbance in the human microbiota 5 Microbiota Collective ecosystem of microorganisms that inhabit the human body 1 Culture-independent Experimental scientific techniques that do not require cultivation of microbes (i.e. PCR, 6 techniques DNA sequencing) Community-state type Vaginal microbiota profile based on species of Lactobacillus that dominates 1 Lipopolysaccharide (LPS) LPS, also known as endotoxin is a component of the outer membrane of Gram- 7 negative bacteria Toll-like receptors (TLRs) Toll-like receptors are pattern recognition receptors (PRRs) that sense invading 8 pathogens or endogenous damage signals Aerobic vaginitis Aerobic vaginitis (AV) is a state of vaginal dysbiosis that is distinct from bacterial 9 vaginosis (BV) Cytolytic vaginosis Lactobacillus overgrowth syndrome resulting in lysis of vaginal epithelial cells 10 Quantitative real-time Molecular technique that is used to quantify the number of copies of the gene of 11 PCR interest in a community sample (i.e. vaginal, gut) Enzyme-linked Immunological assay used to measure antibodies, proteins, antigens and glycoproteins 12 immunosorbent assays in biological samples (ELISA) IL-1β Pro-inflammatory cytokine associated with stress,injury, inflammation and infection 14 Bacteriocins Antimicrobial compounds produced by bacteria that kill or inhibit other bacteria 15 Table 1: Glossary of terms Invivo Healthcare, The New Warehouse, Libby’s Drive, Stroud, GL5 1RN page 2 Please call us on 0333 241 2997, or visit us at invivohealthcare.com INTERPRETIVE GUIDE BACKGROUND Vaginal Microbiota The human body is colonised by a multitude of microorganisms collectively referred to as the human microbiota. In females, one of the main microbiota sites is the genital tract that hosts over 200 bacterial species. Unlike other microbiota sites, the vagina is extremely dynamic and is affected by numerous host factors including age, ethnic background, sexual intercourse, changes in hormone levels during pregnancy, menstruation and menopause, genital infections and personal hygiene16. Due to its extensive interaction and cross-talk with host immune responses, small fluctuations in the vaginal microbiota can impact vaginal health significantly. The disruption of homeostasis, termed dysbiosis, has been linked to a multitude of conditions, including bacterial vaginosis (BV), miscarriages, preterm delivery, pelvic inflammatory disease (PID) and increased risk of sexually transmitted infections (STIs). Homeostasis Detailed composition of the vaginal microbiota has been defined through high-throughput 16S rRNA gene sequencing. A cross-sectional study using 394 healthy women from four ethnic groups classified the vaginal microbiota into five microbial communities, termed community state types (CST), which are generally dominated by Lactobacillus spp.1. Briefly, CST I, II, III and V are dominated by Lactobacillus species (Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners and Lactobacillus jensenii, respectively) and CST IV has no dominant species and is classified as the diverse group, with higher numbers of strictly anaerobic bacteria and BV-associated bacteria such as Prevotella, Gardnerella, Aerococcus, Finegoldia, and Mobiluncus 1. Lactobacilli are extremely important for vaginal health due to their protective and antimicrobial functions. Lactobacilli produce lactic acid, creating an acidic environment (pH 2.8–4.2) that is inhospitable to many non-Lactobacillus commensals and potential vaginal pathogens17,18. For example, reduced infectivity of Chlamydia trachomatis by L. crispatus, via lactic acid production and acidic pH, has been reported in vitro19. Lactic acid also induces autophagy in epithelial cells to degrade intracellular microorganisms and promote homeostasis20. In addition, some Lactobacillus species also produce antimicrobial compounds such as bacteriocins that inhibit growth of pathogenic microorganisms such as Candida albicans, Prevotella bivia and E. coli21. In addition to their direct role in regulating vaginal health, Lactobacilli also modulate the host immune response, mediated through vaginal epithelial cells22,23. Lactobacilli are tolerated by vaginal epithelial cells and inhibit induction of proinflammatory cytokines such as IL-6, IL-1β and TNF-α24. Vaginal lactic acid has also been shown to mediate an antiinflammatory response in the presence of inflammatory inducing pathogens25. Dominance of Lactobacillus spp. is therefore a good marker for vaginal health and indicates functional host-microbial interactions. Dysbiosis Deviations from a healthy microbiota composition or homeostasis, termed dysbiosis, has been linked to pelvic inflammatory disease26 preterm labour27 and miscarriages28. Vaginal dysbiosis is often defined as a prolonged deviation from a low-diversity, Lactobacilli-dominated vaginal microbiota29. The most common dysbiosis of the vaginal microbiota is bacterial vaginosis (BV), an anaerobic polymicrobial dysbiosis29,30. BV-associated dysbiosis typically includes increased Gardnerella vaginalis, Atopobium vaginae and other anaerobes including Megasphaera spp., Dialister spp., Prevotella spp., Mobiluncus spp. with, or without, a low relative abundance of L. iners31,32. CST IV microbiota profiles show increased dominance of these BV-associated anaerobic bacteria. However, as the CST IV profile is detected in healthy women, recent studies now support a role for polymicrobial interactions, biofilm formation and host immune responses in the aetiology of BV5,33. Clinical signs of BV include malodour, a creamy grey discharge and an elevated vaginal pH, linked to reduced lactic acid- production31. Malodour, has been linked to increased vaginal biogenic amines (BAs), including putrescine, cadaverine, and trimethylamine34, that are often produced by BV-associated anaerobic bacteria. In addition, BV associated bacteria produce little lactic acid, but produce several immunomodulatory substances, including succinate, sialydases and proteases, as well as proinflammatory substances such as lipopolysaccharides
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