Amsel's Clinical Criteria Research Articles

Integrating compositional and functional content to describe vaginal microbiomes in health and disease

BackgroundA Lactobacillus-dominated vaginal microbiome provides the first line of defense against adverse genital tract health outcomes. However, there is limited understanding of the mechanisms by which the vaginal microbiome modulates protection, as prior work mostly described its composition through morphologic assessment and marker gene sequencing methods that do not capture functional information. To address this gap, we developed metagenomic community state types (mgCSTs) which use metagenomic sequences to describe and define vaginal microbiomes based on both composition and functional potential.ResultsMgCSTs are categories of microbiomes classified using taxonomy and the functional potential encoded in their metagenomes. MgCSTs reflect unique combinations of metagenomic subspecies (mgSs), which are assemblages of bacterial strains of the same species, within a microbiome. We demonstrate that mgCSTs are associated with demographics such as age and race, as well as vaginal pH and Gram stain assessment of vaginal smears. Importantly, these associations varied between mgCSTs predominated by the same bacterial species. A subset of mgCSTs, including three of the six predominated by Gardnerellavaginalis mgSs, as well as mgSs of L. iners, were associated with a greater likelihood of bacterial vaginosis diagnosed by Amsel clinical criteria. This L. iners mgSs, among other functional features, encoded enhanced genetic capabilities for epithelial cell attachment that could facilitate cytotoxin-mediated cell lysis. Finally, we report a mgSs and mgCST classifier for which source code is provided and may be adapted for use by the microbiome research community.ConclusionsMgCSTs are a novel and easily implemented approach to reduce the dimension of complex metagenomic datasets while maintaining their functional uniqueness. MgCSTs enable the investigation of multiple strains of the same species and the functional diversity in that species. Future investigations of functional diversity may be key to unraveling the pathways by which the vaginal microbiome modulates the protection of the genital tract. Importantly, our findings support the hypothesis that functional differences between vaginal microbiomes, including those that may look compositionally similar, are critical considerations in vaginal health. Ultimately, mgCSTs may lead to novel hypotheses concerning the role of the vaginal microbiome in promoting health and disease, and identify targets for novel prognostic, diagnostic, and therapeutic strategies to improve women’s genital health.6QiSoF_fVWyny-AcPZFxWyVideo

Efficacy and safety of the combination nifuratel-nystatin and clindamycin-clotrimazole, in the treatment of bacterial vaginosis. Randomized controlled clinical trial

Introduction: Bacterial vaginosis (BV) is the most frequent gynecological infection in women of reproductive age, thus driving the search for effective and safe treatments. Objective: To compare the efficacy and safety of the combination nifuratel-nystatin and clindamycinclotrimazole, in the treatment of bacterial vaginosis. Materials and Methods: Randomized controlled clinical trial in 147 single women (18 to 39 years old), non-pregnant and sexually active, with a diagnosis of BV according to the Amsel clinical criteria and the Nugent score; between 2016 and 2018. In a highly complex private clinic in Armenia, Colombia. Women were randomized into two groups: "A" (73 participants: nifuratel (500 mg) - nystatin (100,000 IU) and "B" (74 participants: clindamycin (100 mg) - clotrimazole (200 mg)); both groups were treated with vaginal ovules, "A" for six days and "B" for three days. All participants were followed-up for clinical and microbiological healing at 7 and 30 days, respectively, after completion of treatment, the STATA® 14.0 program was used. Results: The mean age of women was 28.35±5.79 years. The clinical cure rate with the nifuratelnystatin combination was 93.15%, and that of clindamycin-clotrimazole 97.29%, (p=0.123). The microbiological cure rate with the nifuratel-nystatin combination was 87.67%, and that of clindamycin-clotrimazole 93.24%, (p=0.102). Regarding safety, there were also no significant differences between the two groups (p=0.144); Mild adverse reactions were observed. Recurrence in group "A" was 12.32% compared to 6.75% in group "B" (p>0.05). Conclusions: In this study, the combinations nifuratel (500 mg) -nystatin (100,000 IU) and clindamycin (100 mg)-clotrimazole (200 mg), reported that both are equally effective and safe options in the treatment of BV. It is necessary to evaluate the effect and safety of other combinations in order to implement timely interventions. Keywords: Vaginosis; Bacterial; Efficacy; Safety; Nifuratel; Nystatin; Clindamycin; Clotrimazole.

Clinico-microbiological retrospective study of abnormal vaginal discharge in women coming to obstetrics and gynecology out patient department at a tertiary care hospital

Introduction: Vaginal discharge is a normal physiological discharge which is usually white or clear, non-offensive and varies with menstrual cycle. Abnormal vaginal discharge if untreated can lead to cervicitis, pelvic inflammatory disease, urinary tract infection and also infertility. As abnormal vaginal discharge presents vaguely in a clinical set up, diagnosis is made by laboratory confirmation for rapid identification of causative pathogens and appropriate treatment. Materials and Methods: This is a retrospective study where records of clinically suspected vaginitis/cervicitis cases from Jan 2016 till June 2018 were studied. History, general examination findings, per speculum and pelvic examination findings, investigations performed and treatment given were collected for these cases and the data was analyzed. The amount, odor, color, pH and consistency of vaginal discharge were noted. Three high vaginal swabs were taken from every patient using sterile swab sticks and labeled. The tests include: saline wet mount, KOH mount and Grams stain. Bacterial vaginosis was diagnosed using the Amsel's clinical criteria and standard microbiological techniques. The collected samples were also cultured on MacConkey’s agar, Blood agar and Sabouraud’s Dextrose agar for aerobic bacterial and fungal cultures respectively. The antibiotic sensitivity testing of all isolates was performed by Kirby Bauer’s disc diffusion method on Muller Hinton agar and interpreted as per CLSI guidelines. Results: This is a retrospective study where records of clinically suspected vaginitis/cervicitis cases were studied. A total of 1680 clinically suspected vaginitis/cervicitis case records were collected in this study. Out of 1680 cases, only 475 (28.27%) cases were positive for growth of a pathogen and the remaining 1205 (71.72%) cases either showed no growth or no pathogens were isolated. Bacterial vaginosis was diagnosed in 120 (7.14%) cases and trichomoniasis was diagnosed in 51 (3.03%) cases. Out of the 475 samples positive for growth, most common isolate was Escherichia coli (195, 41.05%), followed by Coagulase negative staphylococcus (130, 27.36%). Conclusion: Escherichia coli was the most common organism causing infection in our study population followed by Coagulase negative staphylococcus. Clinical diagnosis along with complete microbiological evaluation is necessary to determine the causative agents of vaginal infections for appropriate management. Keywords: Abnormal vaginal discharge, Vaginitis, Cervicitis, High vaginal swabs.

Prevalence of bacterial vaginosis among antenatal attendees with abnormal vaginal discharge in a secondary health facility in Delta State, Nigeria

Introduction: Bacterial vaginosis (BV) is the most common lower genital tract syndrome among women of reproductive age with significant adverse outcome in pregnancy. Routine screening of antenatal patients for BV is not recommended, however, it is important to determine its prevalence especially in pregnant women who complain of abnormal vaginal discharge. Methods: This was a cross-sectional study of 340 antenatal attendees who complained of an abnormal vaginal discharge. The high vaginal swab was collected from each woman and subjected to wet film and Gram reaction. The diagnosis of BV was made using the Amsel's clinical criteria. Data were analyzed using the EPI-INFO statistical package and the results were presented as percentages and proportions. Results: BV was detected in 105 pregnant women with abnormal vaginal discharge, therefore, giving a prevalence rate of 30.4%. Positive correlates of BV included multiple sexual partners (P = 0.0001) and cigarette smoking (P = 0.008). Conclusion: BV is a prevalent disorder in pregnant women with abnormal vaginal discharge.

The Effect of Prangos Ferulacea Vaginal Cream on Accelerating the Recovery of Bacterial Vaginosis: A Randomized Controlled Clinical Trial.

The present study was conducted on the effect of prangos ferulacea vaginal cream on accelerating the recovery of bacterial vaginosis. This randomized controlled clinical trial was conducted on 100 non-pregnant women referring to health centers affiliated to Lorestan University of Medical Sciences in 2016 with the diagnosis of bacterial vaginosis based on the patient's complaints, Amsel's clinical criteria, and the Nugent microscopic criteria. The women were randomly divided into two groups of 50. One group was treated with oral metronidazole plus Prangos ferulacea vaginal cream and the other with oral metronidazole plus a placebo vaginal cream for seven days. The patient's complaints, Amsel's clinical criteria and the Nugent microscopic criteria were assessed seven days after treatment. The data were analyzed using SPSS, version 20, with a significance level of 0.05. The response to oral metronidazole plus Prangos ferulacea vaginal cream treatment was 94% according to Amsel's clinical criteria and 88% according to the Nugent microscopic criteria. The response to oral metronidazole plus placebo vaginal cream treatment was 94% according to Amsel's criteria and 86% according to the Nugent criteria. The analysis of the patients' complaints, Amsel's clinical criteria and the Nugent microscopic criteria showed significant differences in each group before and after the treatment. This trial showed that Prangos ferulacea vaginal cream accelerated the recovery of bacterial vaginosis of patients with bacterial vaginosis. It can be used effectively as a complementary treatment with oral metronidazole in cases of medication resistance and also in people wishing to use herbal remedies Trial Registration Number: IRCT2016042327534N1.

Role of Gardnerella Vaginalis Among Symptomatic Bacterial Vaginosis Patients Attending at Chittagong Medical College Hospital

Background : Now a day’s bacterial vaginosis is an extremely common health problem for women in the world which causes many complications both in the pregnancy and non-pregnancy states. G. vaginalis is most important cause of bacterial vaginosis. Materials and methods : A cross sectional study was conducted to assess the role of Gardnerella vaginalis as an etiological agent of bacterial vaginosis. This study was undertaken to assess the patients attending at the outpatient department of Gynaecology and Obstetrics of Chittagong Medical College Hospital, Chittagong. A total of 170 sexually active female in the age group of 15-45 years, with abnormal vaginal discharge were selected for the study. A detailed history and a thorough clinical examination of all the cases were done. pH of the vaginal discharge was measured and three high vaginal swabs were collected. Bacterial vaginosis was diagnosed using Amsel’s criteria and Nugent’s method. Gardnerella vaginalis was isolated and identified with their drug sensitivity test by standard methods. Results : In this study 38(22.35%) Gardnerella vaginalis were isolated by culture and bacterial vaginosis was detected by Amsel clinical criteria (Clinical method) 43(25.30%), Gram stain Nugent criteria (Gold Standard) 45(26.47%).Our study showed a relatively high prevalence of bacterial vaginosis and high isolation rate of Gardnerella vaginalis within the positive bacterial vaginosis patients. Conclusion : This result helps proper management and treatment.
 JCMCTA 2017 ; 28 (2) : 81-87

Anaerobic bacteriological profile of leukorrhea in reproductive age group women

INTRODUCTION: Leukorrhea is one of the most common complaints of women in their reproductive age group attending the OBG OPD and Bacterial Vaginosis (BV) is its most common cause. AIM: of the study was to isolate and identify the anaerobic bacteria causing leukorrhea. MATERIALS AND METHODS: This was a cross sectional study conducted in a medical college and tertiary care hospital, on 250 married women in their reproductive age group attending the OBG OPD with complaints of leukorrhea. High vaginal swabs were collected from these women and BV was diagnosed using Amsel's clinical composite criteria. Anaerobic bacteria were isolated from the samples using standard operative procedure. RESULTS: Out of 250 women, 220 (88%) had a thin, grey, homogenous discharge and 30 (12%) had thick, white non offensive discharge, indicating vaginal candidiasis. Out of the 220 women, 123 (55.9%) were diagnosed to have BV by Amsel's criteria and 42 (19%) of them were culture positive for anaerobic bacteria. Porphyromonas spp. (55%) was the most common bacteria isolated, followed by Peptostreptococcus spp. (17%) and Bacteroides spp. (14%). CONCLUSION: Anaerobic bacteria are important pathogens in the causation of bacterial vaginosis along with other aerobic organisms.

Detection of Bacterial Vaginosis and Comparison Of Clinical & Microbiological Methods Among Women Attending Chittagong Medical College Hospital

Bacterial vaginosis is a special public health concern in the world because of high burden of reproductive and pregnancy related morbidity. This present study was undertaken to detect the bacterial vaginosis in reproductive women by different clinical and microbiological methods on patients attending at the out patient department of Gynaecology and Obstetrics of Chittagong Medical College Hospital. A total of 170 sexually active female in the age group of 15-45 years, with abnormal vaginal discharge were selected for the study. In this study bacterial vaginosis was detected by Amsel clinical criteria (Clinical method) Gram stain Nugent criteria (Gold Standard) culture and by newly developed BV assay test and compared these methods. Out of 170 study cases, 43(25.30%) cases were diagnosed as bacterial vaginosis by Amsel criteria, 45(26.47%) cases were positive by Nugent criteria, 46(27.06%) cases were positive by BV assay test and 38(22.35%) cases were culture positive for Gardnerella vaginalis. Sensitivity of the clinical criteria (Amsel) BV assay test, and culture were 95.5%, 97.8% and 84.4% respectively in response to Gold standard Nugent criteria. All these findings make the BV assay test more accurate and reliable screening procedure for the diagnosis of bacterial vaginosis which help our physicians and patients.
 JCMCTA 2016 ; 27 (1) : 40 - 45

Vaginosis bacteriana

Bacterial vaginosis (BV) is the main cause of vaginal dysbacteriosis in the women during the reproductive age. It is an entity in which many studies have focused for years and which is still open for discussion topics. This is due to the diversity of microorganisms that cause it and therefore, its difficult treatment. Bacterial vaginosis is probably the result of vaginal colonization by complex bacterial communities, many of them non-cultivable and with interdependent metabolism where anaerobic populations most likely play an important role in its pathogenesis. The main symptoms are an increase of vaginal discharge and the unpleasant smell of it. It can lead to serious consequences for women, such as an increased risk of contracting sexually transmitted infections including human immunodeficiency virus and upper genital tract and pregnancy complications. Gram stain is the gold standard for microbiological diagnosis of BV, but can also be diagnosed using the Amsel clinical criteria. It should not be considered a sexually transmitted disease but it is highly related to sex. Recurrence is the main problem of medical treatment. Apart from BV, there are other dysbacteriosis less characterized like aerobic vaginitis of which further studies are coming slowly but are achieving more attention and consensus among specialists.

Diagnosis of Bacterial Vaginosis: Amsel’s Criteria vs Nugent’s scoring

Bacterial Vaginosis (BV) is the most common cause of vaginitis. If untreated, it is associated with serious complications. BV is a poly-microbial, synergistic infection, involving over-growth of multiple bacterial pathogens. Vaginal flora becomes altered leading to increased pH due to decrease in lactobacilli (which produce H2O2).Although Nugent's criterion is considered as the gold standard in diagnosis of BV, routinely a combination of various methods is used for the diagnosis of bacterial vaginosis(BV). In the present study we compared Amsel's composite clinical criteria with Nugent's method for the diagnosis of BV. The study was undertaken from February - November, 2015 at a Tertiary Care Hospital in Hyderabad. The study consisted of women with complaints of vaginal discharge. BV was diagnosed based on Amsel's criteria and Nugent's scoring. For each case, the following were interpreted: pH, Whiff test (a test in which vaginal secretions are mixed with 10% KOH resulting in a fishy odor typical of bacterial vaginosis)and the presence of the clue cells on vaginal wet smear. Gram staining was performed for Nugent's method. The present study included 362 cases of abnormal vaginal discharge. Prevalence of BV was 48%. Age group 24-29 years was most affected. Amsel's criteria detected 170/362 whereas Nugent score identified 173/362 subjects as having bacterial vaginosis. In comparison with Nugent's criteria the sensitivity, specificity, positive predictive value and negative predictive value of Amsel's criteria were 78.72%, 92.35%, 75.51% and 93.54%. The prevalence rate of BV in present study high. With limited resources in developing countries like ours, there is a great need for inexpensive diagnostic methods for bacterial vaginosis. Amsel's criteria is as good as Nugent's scoring in diagnosis of BVand it is simple, easy, cost effective, fast and reliable, and can be done in OPD which can be used for precise and fast treatment. Keywords Bacterial vaginosis, Amsel's criteria, Nugents scoring, Clue cells, Whiff test, Vaginal Discharge

Diagnostic value of Amsel's clinical criteria for diagnosis of bacterial vaginosis.

Introduction:Bacterial vaginosis (BV) is one of the most prevalent infections in women of reproductive age. Amsel’s criteria and Nugent scoring system are among the most commonly used diagnostic methods. Although Nugent scoring system is considered the gold standard for diagnosing BV, it is time consuming and costly, and its interpretation needs lab equipment and experts. Hence, most physicians are inclined to use simpler clinical criteria that are yet accurate instead. The present study aimed to determine the diagnostic value of Amsel’s criteria in diagnosing BV.Material and Methods:This present study was conducted to validate diagnostic tests of BV in 120 married women in 2013. Amsel’s criteria and Nugent scoring system were used to diagnose BV. Nugent scoring system was considered the gold standard and sensitivity, specificity, positive predictive value and negative predictive value of Amsel’s criteria were compared with those of Nugent scoring system.Results:Kappa coefficient was used to assess the diagnostic value of Nugent scoring system and Amsel’s criteria. Kappa coefficient was found 0.8, which confirms the reliability of both diagnostic methods. McNemar test did not reveal a significant difference between Nugent scoring system and Amsel’s criteria in terms of diagnosing BV. As compared to Nugent scoring system, Asmel’s criteria enjoy sensitivity of 0.91, specificity of 0.91, positive predictive value of 0.86, negative predictive value of 0.94, and accuracy of 0.91.Conclusion:If lab equipment is not available for diagnosing BV, Amsel’s criteria can be as good as Nugent scoring system at diagnosing this infection.

Comparing the Therapeutic Effects of Garlic Tablet and Oral Metronidazole on Bacterial Vaginosis: A Randomized Controlled Clinical Trial

Background:Bacterial vaginosis (BV) is one of the most common gynecological infections during reproductive age. Although metronidazole is one of the most effective medications recommended as the first-line treatment, it has various side effects. Because of the side effects and contraindications of some chemical medicines, using herbs has been investigated in treating BV.Objectives:The aim of this study was to compare the effect of garlic tablet (Garsin) and oral metronidazole in clinical treatment of the BV in women referred to Resalat Health Center, affiliated with Mazandaran University of Medical Sciences, in 2013.Patients and Methods:This randomized clinical trial was conducted on 120 married women aged 18 to 44 years who were diagnosed with BV by Amsel’s clinical criteria and Gram staining. Enrolled women were randomly allocated to two groups of 60 patients and were treated with either garlic tablet or oral metronidazole for seven days. Amsel’s criteria and Gram stain were assessed seven to ten days after beginning the treatment period and side effects were registered.Results:Amsel’s criteria were significantly decreased after treatment with garlic or metronidazole (70% and 48.3%, respectively; P < 0.001). Therapeutic effects of garlic on BV were similar to that of metronidazole (63.3% and 48.3%, respectively; P = 0.141). There were significant differences between the two treatment groups in terms of side effects; metronidazole was associated with more complications (P = 0.032).Conclusions:This study reveals that garlic could be a suitable alternative for metronidazole in treatment of BV in those interested in herbal medicines or those affected by side effects of metronidazole.

Prevalence and epidemiological correlates of bacterial vaginosis among nonpregnant females at a tertiary care center in Assam, India

Purpose: Bacterial vaginosis is the most common type of vaginal infection among women of reproductive age and accounted for at least one-third of all vulvovaginal infections. The main aim of this prospective study was to determine the risk factors that may be associated with the occurrence of bacterial vaginosis among the reproductive age group females in Assam. Materials and Methods: A total of two hundred married, nonpregnant females in the reproductive age group who complained of one or more symptoms suggesting lower genital tract infection were enrolled in this study. Detailed history of the patients was taken. Three high vaginal swabs were obtained after informed consent of the patients. Bacterial vaginosis was diagnosed on the basis of Amsel's clinical criteria and Nugent's Gram stain scoring method. Data was statistically analyzed using Chi-square test. Results: Our study found prevalence of bacterial vaginosis to be 51.5% (103/200) by Amsel's criteria and 49% (98/200) by Nugent's criteria respectively. Significant association was found between bacterial vaginosis and age, socioeconomic status, parity and contraceptives. However, no significant association was found between duration of marriage and stage of the menstrual cycle. Conclusion: A relatively higher prevalence rate of bacterial vaginosis was found in the population under study, which may be attributed to factors like low socioeconomic status and unhygienic practices. Barrier contraception was found to be an effective method for prevention of transmission of bacterial vaginosis. However, more studies may be needed to confirm and evaluate the causes of these associations, so that a preventive strategy can be planned.

Comparison of Nucleic Acid Amplification Assays with BD Affirm VPIII for Diagnosis of Vaginitis in Symptomatic Women

A commercially available, nonamplified, nucleic acid probe-based test system (BD Affirm VPIII) was compared with nucleic acid amplification (NAA)-based assays for determining the etiology of vaginitis in a cohort of 323 symptomatic women. First, a semiquantitative, multiplexed PCR assay (BV-PCR) and the Affirm VPIII Gardnerellavaginalis test were compared with a unified bacterial-vaginosis (BV) reference standard incorporating both Nugent Gram stain scores and Amsel clinical criteria. In the evaluable population of 305 patients, BV-PCR was 96.9% (191/197) sensitive and 92.6% specific (100/108) for BV, while Affirm VPIII was 90.1% sensitive (179/197) and 67.6% specific (73/108). Second, a multiplexed PCR assay detecting Candida albicans and Candida glabrata (CAN-PCR) was compared with the Affirm VPIII Candida test using a reference standard for vulvovaginal candidiasis (VVC) of yeast culture plus exclusion of alternate vaginitis etiologies. In the population evaluated (n = 102), CAN-PCR was 97.7% sensitive (42/43) and 93.2% specific (55/59) and Affirm VP III was 58.1% sensitive (25/43) and 100% specific (59/59) for VVC. Finally, the results of a commercial NAA test (GenProbe Aptima Trichomonas vaginalis assay; ATV) for T. vaginalis were compared with the Affirm VPIII Trichomonas vaginalis test. In the absence of an independent reference standard for trichomonal vaginitis (TV), a positive result in either assay was deemed to represent true infection. In the evaluable cohort of 388 patients, the sensitivity of ATV was 98.1% (53/54) versus 46.3% (25/54) for Affirm VPIII. The diagnostic accuracy of the combined NAA-based test construct was approximately 20 to 25% higher than that of the Affirm VPIII when modeled in populations with various prevalences of infectious vaginitis.

Bacterial Communities in Women with Bacterial Vaginosis: High Resolution Phylogenetic Analyses Reveal Relationships of Microbiota to Clinical Criteria

BackgroundBacterial vaginosis (BV) is a common condition that is associated with numerous adverse health outcomes and is characterized by poorly understood changes in the vaginal microbiota. We sought to describe the composition and diversity of the vaginal bacterial biota in women with BV using deep sequencing of the 16S rRNA gene coupled with species-level taxonomic identification. We investigated the associations between the presence of individual bacterial species and clinical diagnostic characteristics of BV.Methodology/Principal FindingsBroad-range 16S rRNA gene PCR and pyrosequencing were performed on vaginal swabs from 220 women with and without BV. BV was assessed by Amsel’s clinical criteria and confirmed by Gram stain. Taxonomic classification was performed using phylogenetic placement tools that assigned 99% of query sequence reads to the species level. Women with BV had heterogeneous vaginal bacterial communities that were usually not dominated by a single taxon. In the absence of BV, vaginal bacterial communities were dominated by either Lactobacillus crispatus or Lactobacillus iners. Leptotrichia amnionii and Eggerthella sp. were the only two BV-associated bacteria (BVABs) significantly associated with each of the four Amsel’s criteria. Co-occurrence analysis revealed the presence of several sub-groups of BVABs suggesting metabolic co-dependencies. Greater abundance of several BVABs was observed in Black women without BV.Conclusions/SignificanceThe human vaginal bacterial biota is heterogeneous and marked by greater species richness and diversity in women with BV; no species is universally present. Different bacterial species have different associations with the four clinical criteria, which may account for discrepancies often observed between Amsel and Nugent (Gram stain) diagnostic criteria. Several BVABs exhibited race-dependent prevalence when analyzed in separate groups by BV status which may contribute to increased incidence of BV in Black women. Tools developed in this project can be used to study microbial ecology in diverse settings at high resolution.

Development and Validation of a Semiquantitative, Multitarget PCR Assay for Diagnosis of Bacterial Vaginosis

Quantitative PCR assays were developed for 4 organisms reported previously to be useful positive indicators for the diagnosis of bacterial vaginosis (BV)--Atopobium vaginae, Bacterial Vaginosis-Associated Bacterium 2 (BVAB-2), Gardnerella vaginalis, and Megasphaera-1--and a single organism (Lactobacillus crispatus) that has been implicated as a negative indicator for BV. Vaginal samples (n = 169), classified as positive (n = 108) or negative (n = 61) for BV based on a combination of the Nugent Gram stain score and Amsel clinical criteria, were analyzed for the presence and quantity of each of the marker organisms, and the results were used to construct a semiquantitative, multiplex PCR assay for BV based on detection of 3 positive indicator organisms (A. vaginae, BVAB-2, and Megasphaera-1) and classification of samples using a combinatorial scoring system. The prototype BV PCR assay was then used to analyze the 169-member developmental sample set and, in a prospective, blinded manner, an additional 227 BV-classified vaginal samples (110 BV-positive samples and 117 BV-negative samples). The BV PCR assay demonstrated a sensitivity of 96.7% (202/209), a specificity of 92.2% (153/166), a positive predictive value of 94.0%, and a negative predictive value of 95.6%, with 21 samples (5.3%) classified as indeterminate for BV. This assay provides a reproducible and objective means of evaluating critical components of the vaginal microflora in women with signs and symptoms of vaginitis and is comparable in diagnostic accuracy to the conventional gold standard for diagnosis of BV.

P3-S7.02 Performance of a point-of-care diagnostic for Bacterial Vaginosis among young reproductive age women in Mysore, India

BackgroundBacterial vaginosis (BV) remains the most common cause of abnormal vaginal discharge in Indian women of reproductive age and is associated with increased susceptibility to HIV/STI and preterm delivery. Diagnosis...

A case control study of anovaginal distance and bacterial vaginosis

We aimed to test the hypothesis that a short anovaginal distance may increase the risk of bacterial vaginosis (BV) due to faecal contamination and disruption of the vaginal microbiota. Women attending two sexually transmitted infection (STI) clinics in Baltimore, Maryland, USA, who complained of a vaginal discharge were asked to participate in a study to measure mucosal immune responses. In this pilot study of all enrolled women, a small plastic ruler was used to measure the anatomic distance from the posterior fourchette to the anus with the participant in the lithotomy position. Cases of BV, defined by Amsel's clinical criteria (n = 62), were compared with controls (n = 31) without BV. We used linear and logistic regression models to adjust for potential confounders. A total of 93 women were recruited (median age 28.6 years, 93% black, 4.4% gonorrhoea infection, 7.4% chlamydia infection, 8.6% trichomonas infection, 67% BV diagnosed). Mean anovaginal distance was 3.22 cm (SD: 0.74, range 1.8-5.2) for controls and 3.37 cm (SD: 0.76, range: 1.8-5.7) for cases (P = 0.38). There was no difference between cases and controls when comparing median values, quartiles and after adjusting for potential confounders. Among high-risk women with multiple co-infections, there was no association between anovaginal distance and clinical diagnosis of BV.

Changes in Vaginal Bacterial Concentrations with Intravaginal Metronidazole Therapy for Bacterial Vaginosis as Assessed by Quantitative PCR

Several fastidious bacteria have been associated with bacterial vaginosis (BV) using broad-range bacterial PCR methods such as consensus sequence 16S rRNA gene PCR, but their role in BV remains poorly defined. We describe changes in vaginal bacterial concentrations following metronidazole therapy for BV. Vaginal swabs were collected from women with BV diagnosed using Amsel clinical criteria, and vaginal fluid was assessed by Gram stain to generate Nugent scores. Follow-up swabs were collected 1 month after a 5-day course of vaginal 0.75% metronidazole gel and analyzed for 24 subjects with cured BV and 24 subjects with persistent BV. Changes in bacterial concentrations were measured using eight bacterium-specific 16S rRNA gene quantitative PCR assays. DNA from several fastidious BV-associated bacteria (BVAB) were present at high concentrations in the vagina prior to treatment. Successful antibiotic therapy resulted in 3- to 4-log reductions in median bacterial loads of BVAB1 (P=0.02), BVAB2 (P=0.0004), BVAB3 (P=0.03), a Megasphaera-like bacterium (P<0.0001), Atopobium species (P<0.0001), Leptotrichia/Sneathia species (P=0.0002), and Gardnerella vaginalis (P<0.0001). Median posttreatment bacterial levels did not change significantly in subjects with persistent BV except for a decline in levels of BVAB3. The presence or absence of BV is reflected by vaginal concentrations of BV-associated bacteria such as BVAB1, BVAB2, Leptotrichia/Sneathia species, Atopobium species, Gardnerella vaginalis, and a Megasphaera-like bacterium, suggesting that these bacteria play an important role in BV pathogenesis and may be suitable markers of disease and treatment response.

Targeted PCR for Detection of Vaginal Bacteria Associated with Bacterial Vaginosis

Several novel bacterial species have been detected in subjects with bacterial vaginosis (BV) by using broad-range PCR assays, but this approach is insensitive for detecting minority species. We developed a series of taxon-directed 16S rRNA gene PCR assays for more sensitive detection of key vaginal bacteria. We sought to determine the prevalence of each species in the vagina, its association with BV, and the utility of PCR for the microbiological diagnosis of BV. Targeted PCR assays were developed for 17 vaginal bacterial species and applied to 264 vaginal-fluid samples from 81 subjects with and 183 subjects without BV. The results were compared to those of two widely accepted methods for diagnosing BV, the use of clinical findings (Amsel criteria) and the interpretation of vaginal-fluid Gram stains (Nugent criteria). Leptotrichia/Sneathia, Atopobium vaginae, an Eggerthella-like bacterium, Megasphaera species, and three novel bacteria in the order Clostridiales are among the bacterial species significantly associated with BV. PCR detection of either a Megasphaera species or one of the Clostridiales bacteria yielded a sensitivity of 99% and a specificity of 89% for diagnosis of BV compared to the Amsel clinical criteria and a sensitivity of 95.9% and a specificity of 93.7% compared to the Nugent criteria (Gram stain). PCR detection of one or more fastidious bacterial species is a more reliable indicator of BV than detection of bacteria, such as Gardnerella vaginalis, previously linked to BV, highlighting the potential of PCR for the diagnosis of BV.