scholarly journals Molecular diagnosis of bacterial vaginosis: Prevalence of Gardnerella vaginalis and Atopobium vaginae in pregnant women

2018 ◽  
Vol 146 (7-8) ◽  
pp. 417-421
Author(s):  
Snezana Matic ◽  
Dane Nenadic ◽  
Jelena Cukic ◽  
Zeljko Mijailovic ◽  
Nevena Manojlovic ◽  
...  

Introduction/Objective. Bacterial vaginosis (BV) is defined as disequilibrium of vaginal microbiota due to proliferation of Gram-negative/variable anaerobes and reduction/depletion of vaginal lactobacilli. Difficulties in interpreting microscopically categorized findings in diagnosis of BV need a molecular analysis of bacteria present in vaginal discharge of patients. In this regard, we performed real-time qPCR analysis of vaginal discharge samples with the goal to explore in which extent prevalence and amount of anaerobes, Gardnerella vaginalis and Atopobium vaginae, are related to findings obtained by microscopy. Methods. This study enrolled 111 asymptomatic pregnant women between 24 and 28 weeks of pregnancy. Gram-stained vaginal smears were evaluated microscopically. Afterwards, DNA of bacteria was extracted from Gram slides and real-time qPCR was performed with the aim to detect and quantify G. vaginalis and A. vaginae. Results. The data of our study showed that 53.2% of patients had normal results, while 20.7% and 26.1% of patients had intermediary (IMD) and BV results, respectively. G. vaginalis and A. vaginae were more frequently found in IMD and BV than in healthy patients; also, the average bacterial number of G. vaginalis and A. vaginae were significantly higher in BV and IMD than in the group with normal findings (p = 0.000). Comparing mutual relation of G. vaginalis and A. vaginae, the prevalence and number of G. vaginalis were in all groups significantly higher than A. vaginae. Conclusion. The data of our study have shown that in distinguishing normal from BV findings, quantification of bacteria may be more important than just molecular detection of bacteria.

Author(s):  
Maleeha Aslam ◽  
Rubeena Hafeez ◽  
Shahbaz Aman ◽  
Anjum Azhar ◽  
Durre Shahwar Lone

Objective: To determine the prevalence of Bacterial Vaginosis (BV) in pregnant women using a simple clinical diagnostic approach. Method: The clinical criteria developed by Amsel`s and colleagues were used as a reference standard for accurate diagnosis of Bacterial vaginosis (BV) infection. This is based on the presence of the following clinical signs 1) Homogeneous thin, white vaginal discharge 2) Presence of clue cells (greater than 20%). 3) Positive amine (whiff) test. 4) Vaginal pH > than 4.5. The presence of any three of the four clinical criteria was considered diagnostic for BV. Samples were collected from 75 pregnant females, irrespective of their gestational duration, presenting with vaginal discharge at the antenatal clinic of Jinnah Hospital, Lahore. Results: BV was diagnosed in 14 females giving an incidence of 18.7%. The, most common symptom was a thin white homogeneous vaginal discharge seen in 27(36%) women. The other diagnostic clinical signs observed were the presence of clue cells on wet mount 9(12%), positive `whiff` test 13(17.3%), and elevated pH>4.5 in 18(24%) women. Microscopic analysis of vaginal secretion revealed diminished polymorph epithelial ratio 19(25.3%) and loss of normal vaginal Lactobaclli 22(29.3%). The other associated findings were candidiasis seen in 16(21.3%) women, while Trichomonas infection in only one woman. These findings were confirmed on Gram staining. Gardnerella vaginalis 9(12%) and Gram variable organisms (Mobiluncus) 11(14.7%) were also identified by Gram smear of vaginal discharge. Conclusion: The clinical diagnosis of BV infection can be established by identifying three of Amsel`s four clinical criteria. In most cases it provides a rapid inexpensive and accurate diagnosis. Strategies should be planned to screen and treat women with BV so as to prevent adverse Obstetric outcome associated with it.


2021 ◽  
Vol 15 (09) ◽  
pp. 1293-1298
Author(s):  
Nedzib Numanovic ◽  
Snezana Ribis ◽  
Jelena Cukic ◽  
Dane Nenadic ◽  
Aleksandar Zivanovic ◽  
...  

Introduction: The aim of the study was to investigate prevalence of bacteria most frequently associated with bacterial vaginosis using Amsel’s criteria as well as to quantify these bacteria by real-time PCR and to explore the difference in their quantity between healthy and bacterial vaginosis samples. Methodology: For classification of vaginal discharge samples Amsel’s criteria have been used. To detect and quantify Gardnerella vaginalis Atopobium vaginae, Lactobacillus spp. and total vaginal microbiome, real-time PCR has been applied. Results: According to results of our study Amsel’s criteria matched well with real-time PCR diversification of healthy women and women with BV. Nevertheless, real-time PCR has been more sensitive in diagnosis of bacterial vaginosis. DNA quantification of bacteria demonstrated that mutual abundance of G.vaginalis and A. vaginae was good bacterial vaginosis marker . On the contrary, Lactobacillus spp. was present in high amount in both healthy and bacterial vaginosis samples, but ratio of investigated bacteria was different between them. In fact, G. vaginalis and A. vaginae comprised only 0.1% of total microbiome in healthy, whereas Lactobacillus spp. took 99.3% of it. Nonetheless, in bacterial vaginosis, G. vaginalis and A. vaginae made up 34.4% of total microbiome, while Lactobacillus spp. was 21.6%. Conclusions: According to the results of our study real-time PCR analysis was more sensitive in diagnosis of bacterial vaginosis than Amsel’s method, as well as it represented fine tool in making a difference between microbial entities in healthy and bacterial vaginosis samples.


2018 ◽  
Vol 32 (4) ◽  
Author(s):  
Giorgio Dirani ◽  
Silvia Zannoli ◽  
Maria Federica Pedna ◽  
Francesco Congestrì ◽  
Patrizia Farabegoli ◽  
...  

Background and aims. Bacterial vaginosis (BV) is one the more frequently identified genital syndrome among childbearing aged women. The basic condition that generates this condition is a modification in the vaginal microbiota. The aim of this paper is to briefly review the current status of the art of BV and to report the results of a pilot study performed with an innovative PCR based technique. Materials and Methods. 36 samples of vaginal fluid routinely submitted for the diagnosis of BV to the Unit of Microbiology – GRHL were comparatively evaluated by standard techniques and with the HP-Vaginiti e Vaginosi NLM kit that simultaneously detects in a quantitative way specific DNA from Candida (albicans, glabrata; krusei, tropicalis), Gardnerella vaginalis, Lactobacillus spp. and Atopobium vaginae. Results and conclusions. Candida spp. has been identified in 8 samples with culture and in 15 with the molecular test. 29 G. vaginalis were found by PCR whereas only in 7 samples a specific prescription for this microbe was present (of which 4 positive). A. vaginae has been identified in 20 samples by the molecular approach and Lactobacillus spp. was identified in 19 samples (by culture) and in 32 by PCR. The overall diagnosis of BV was made in 9 patients by standard techniques and in 7 by applying the molecular approach. (Cohen’s kappa test: 0,84). The findings of this study clearly demonstrate that the joint use of the routine culture- based techniques with the multiplex PCR methods amplifies by far the sensitivity of the overall diagnostic workflow of BV.


2017 ◽  
Vol 66 (8) ◽  
pp. 1217-1224 ◽  
Author(s):  
Carolina Sanitá Tafner Ferreira ◽  
Gilbert Gerard Donders ◽  
Cristina Maria Garcia de Lima Parada ◽  
Andrea da Rocha Tristão ◽  
Thaiz Fernandes ◽  
...  

2020 ◽  
pp. 21-24
Author(s):  
A.D. Krutikova

Patients with bacterial vaginosis are diagnosed with chronic generalized inflammatory and inflammatory-dystrophic periodontal diseases, such as chronic catarrhal gingivitis, periodontitis of initial and first severity. The aim of the study was to compare the effectiveness of the treatment of inflammatory and inflammatory-dystrophic periodontal diseases in women with bacterial vaginosis and their sexual partners. Materials and methods. 28 heterosexual couples aged 18-45 years took part in the survey. Clinical dental examination was performed at the time of treatment and 3 months after completion of treatment. Green-Vermilion indices, RMA (in Parma modification), complex periodontal index according to Leus (KPI), Svrakov's number were determined. Both groups of patients were examined for the presence of Gardnerella vaginalis and Atopobium vaginae in the oral cavity by PCR testing. The therapeutic course was the same for all groups of patients. General therapy included: "Fluconazole" 50 mg 1 time per day (7 days), "Clindamycin" 150 mg every 6 hours (5 days), starting from the 3rd day of antibiotics - probiotic "Symbiter acidophilus concentrated "(21 days), Calcium-D3-Nycomed 1 tab. during dinner 30 days. Local treatment included: oral baths of the drug "Stomatophyte" after morning and evening hygienic procedures (7 days), application of "Metrogil-dent" on the gums (7 days), the drug "Lizak" 1 tab. keep in the mouth until complete dissolution every 6 hours (5 days), after the use of "Stomatophyte" and "Metrogil - dent" - application of "Symbiter omega" on the gums in silicone caps at night (21 days). Results. In women, chronic generalized catarrhal gingivitis was detected in 10.71%, chronic generalized periodontitis of the initial degree in 64.29% and chronic generalized periodontitis of the I degree in 25%. In men who are carriers of pathogens of BV, chronic generalized catarrhal gingivitis was found in 7.14%, chronic generalized periodontitis of the initial degree in 60.71% and chronic generalized periodontitis in grade 32.1. General and local dental treatment of women with bacterial vaginosis and their sexual partners showed a positive objective dynamics of dental status. Thus, after 3 months of clinical and laboratory observations, there were no complaints, the state of oral hygiene improved significantly. If before the treatment the Green-Vermilion index in general in women with BV was 1.323 ± 0.035 points, then after 3 months it was equal to 1.032 ± 0.021. A similar dynamics was determined in men: from the starting index of 1.336 ± 0.041, the Green-Vermilion index decreased to 1.048 ± 0.036. 3 months after treatment, the PMA of patients decreased by 49.03%, in men - by 51.95%, no significant difference between the indicators of groups I and II was registered, while the difference between the results in the middle of groups I and II is significant . According to the KPI index, women had a significant difference between baseline and post-treatment outcomes of 1.98 points, while men had a score of 2.01 points. No significant difference was found between the results of groups I and II both before and after treatment. A similar positive dynamics characterizes the indicator - Srakov's number, according to this indicator before treatment and after differences between groups was not detected. The significant difference between the indicators in the group of women and men was 2,789 and 2,831, respectively. Before treatment and after the detection rate in the oral cavity, Gardnerella vaginalis and Atopobium vaginae did not have a significant difference between the group of men and women, but there was a clear difference between the parameters obtained at the initial examination and 3 months after treatment in the middle and second groups. Thus, Gardnerella vaginalis was detected less by 67.87% and 61.59%, Atopobium vaginae - by 65.79% and 58.65% in women and men, respectively. Conclusions. The developed and patented treatment regimen for patients with bacterial vaginosis contributes to the regression of inflammatory phenomena, improves oral hygiene, and is suitable for the treatment of women with this comorbidity and for the treatment of their sexual partners. The dynamics of reducing the percentage of detection of bacterial vaginosis pathogens corresponded to the dynamics of the clinical picture of periodontal disease in both women with bacterial vaginosis and men who were their sexual partners, which once again confirms the equal effectiveness of treatment for both gender groups.


2017 ◽  
Vol 66 (4) ◽  
pp. 57-67 ◽  
Author(s):  
Veronika V. Nazarova ◽  
Elena V. Shipitsyna ◽  
Ekaterina N. Gerasimova ◽  
Alevtina M. Savicheva

Background. Bacterial vaginosis is disturbance of the balance of the vaginal microflora, associated with a number of infectious diseases of the urogenital tract and adverse pregnancy outcomes. In this country, for the detection of vaginal dysbiotic conditions, the test Femoflor-16 (DNA-Technology, Moscow) is widely used, however interpretation algorithms of this test do not include the category of BV. Aim. The study aimed to elaborate diagnostic criteria for the detection of BV using Femoflor-16 test. Materials and methods. Women of reproductive age addressing a gynecologist with vaginal discharge were enrolled in the study. For clinical diagnosis of BV, the Amsel criteria were used, laboratory analysis for BV was performed via microscopic investigation of vaginal discharge using the Nugent score. Samples of vaginal discharge from all women were analyzed with the test Femoflor-16, intended for characterizing vaginal microbiocenosis using multiplex quantitative real-time PCR. Results. A total of 280 women were included in the study. BV was diagnosed in 86 women (31%) using the Amsel criteria, and in 81 women (29%) using the Nugent score. All groups of anaerobic bacteria included in Femoflor-16 test were shown to be associated with BV, with the exception of bacteria of the genus Mobiluncus, which are detected together with phylogenetically related but not BV-associated bacteria of the genus Corynebacterium. A low amount of lactobacilli (< 10% of total bacterial load) coupled with an elevated amount of Gardnerella vaginalis/Prevotella bivia/Porphyromonas (> 1%) and/or Eubacterium (> 2%) and/or Sneathia/Leptotrichia/Fusobacterium (> 0.1%) and/or Megasphaera/Veillonella/Dialister (> 0.1%) and/or Lachnobacterium/Clostridium (> 0.1%) and/or Peptostreptococcus (> 0.1%) and/or Atopobium vaginae (> 0.2%) detected BV with a sensitivity of 99% and specificity of 93%. Conclusions. Criteria for BV diagnosis using the test Femoflor-16 have been elaborated, which enable to detect BV or exclude it with a sensitivity of 99% and specificity of 93%. These criteria for BV and criteria of the test manufacturers for severe anaerobic dysbiosis determine to a large extent the same category of the vaginal microbiocenosis.


2016 ◽  
Vol 65 (4) ◽  
pp. 76-82
Author(s):  
Elena V. Shipitsina ◽  
Tatyana A. Khusnutdinova ◽  
Olga S. Ryzhkova ◽  
Anna A. Krysanova ◽  
Olga V. Budilovskaya ◽  
...  

Introduction. Bacterial vaginosis (BV) is associated with a number of reproductive health disorders, therefore timely and accurate diagnosis of this condition is exceedingly important. Objective.Comparison of effectiveness of clinical and laboratory diagnostics of BV in women with vaginal discharge. Material and methods. In total, 318 patients addressing gynecological clinics with complaints about vaginal discharge participated in the study. Clinical diagnostics of BV was performed in the clinics participating in patient enrollment in accordance with their clinical practice. For laboratory diagnostics, microscopy of Gram stained smears according to the Nugent method and quantitative real-time PCR were used. Sensitivity and specificity of clinical diagnostics of BV and the molecular method were evaluated using the Nugent method as reference standard. Results. With the Nugent method, BV was diagnosed in 27% of women, with real-time PCR — in 37% of women. Using clinical signs of BV, the condition was diagnosed in 91% women. Sensitivity and specificity of the real-time PCR were 97% and 87%, respectively. Sensitivity of clinical diagnostics was 100%, but specificity was only 17%. Conclusions. Diagnostics of BV based only on the presence of vaginal discharge leads to false positive results and requires laboratory confirmation. The molecular method has a high sensitivity and satisfactory specificity for BV diagnosis and can be used as an alternative to the Nugent method.


2021 ◽  
pp. 72-77
Author(s):  
L.V. Kalugina ◽  
T.F. Tatarchuk ◽  
I.V. Shmulian

Research objective: to examine the incidence of asymptomatic and recurrent bacterial vaginosis (BV) in Ukrainian women and to evaluate diagnostic and treatment strategies for the disease control. Materials and methods. This study lasted from January to March 2021 and included 277 doctors of women's clinics, gynecological hospitals and oncology centers from all regions of Ukraine. Information was provided on 12 896 patients between of 18 and 59 ages with BV. Results. Analysis of data from 12 896 questionnaires allowed us to determine that women from 18 to 35 years of age (62.6%) were the most frequently consulted about BV. Only 58.72% patients with BV reported about abnormal vaginal discharge as the reason for visit, while 41.24% of the respondents reported other reasons. Questionnaire analysis of patients with vaginal discharge revealed scant symptoms of the disease: only 62.67% complained about the change of discharge character; 38.49% and 36.35% of examinees pointed to the vaginal itching and unpleasant odor, respectively; 19.89% of patients were troubled by dysuric symptoms and 15.10% of patients mentioned dyspareunia. In the list of submitted questionnaires, 59.92% of patients indicated an additional examination to identify the causes of vaginitis by the PCR method (urogenital scraping). According to its results, Gardnerella vaginalis was detected in diagnostically significant concentrations in 71.49% of patients, Atopobium vaginae was found in 11.32% of cases, Mobiluncus spp. was detected in 19.96% of patients and Trichomonas vaginalis was found in 13.12% of patients. Candida colonization was diagnosed in 62.07%, with Candida albicans in 48,52% cases and non-albicans forms in 13,55% of patients.In the treatment of BV priority was given to the vaginal forms of the most compliant therapy regimens: 2 times a day for 3 days or once a day for 7 days, depending on the clinical situation. Some patients opted for a long course of treatment for recurrent infections, choosing the combination metronidazole 750 mg / miconazole 200 mg 1 suppository once a day for 5 days for 12 months.Conclusion. Diagnosis of BV in Ukraine continues to be based on clinical conditions and requires a unified algorithm that will be based on current clinical guidelines and dictated by national protocols for management. High mycotic colonization (62,07%) in BV makes it expedient to use combined agents (metronidazole 750 mg / miconazole 200 mg tioconazole/tinidazole) both as etiotropic and empirical therapy and especially if there are clinical conditions that may can cause BV (before surgical interventions, after antibiotic therapy, in endocrine pathology, etc.).


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