scholarly journals Quantification of Gardnerella vaginalis, Atopobium vaginae and Lactobacillus spp. in bacterial vaginosis

2021 ◽  
Vol 15 (09) ◽  
pp. 1293-1298
Author(s):  
Nedzib Numanovic ◽  
Snezana Ribis ◽  
Jelena Cukic ◽  
Dane Nenadic ◽  
Aleksandar Zivanovic ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Gardnerella Vaginalis ◽  
Pcr Analysis ◽  
Making A Difference ◽  
The Difference ◽  
Lactobacillus Spp ◽  
Atopobium Vaginae

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.

scholarly journals Bacterial vaginosis: epidemiologic, clinical and diagnostic updates

2018 ◽  
Vol 32 (4) ◽  
Author(s):  
Giorgio Dirani ◽  
Silvia Zannoli ◽  
Maria Federica Pedna ◽  
Francesco Congestrì ◽  
Patrizia Farabegoli ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Current Status ◽  
Gardnerella Vaginalis ◽  
Vaginal Fluid ◽  
Molecular Approach ◽  
Candida Spp ◽  
Kappa Test ◽  
Lactobacillus Spp ◽  
Standard Techniques ◽  
Atopobium Vaginae

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.

Significance of Gardnerella vaginalis genotyping in diagnosis of recurrent bacterial vaginosis

2021 ◽  
Vol 1 (30) ◽  
pp. 48-52
Author(s):  
A. A. Krysanova ◽  
A. E. Gushchin ◽  
A. M. Savicheva
Keyword(s):  
Bacterial Vaginosis ◽  
Real Time ◽  
Real Time Pcr ◽  
Test System ◽  
Reproductive Age ◽  
Gardnerella Vaginalis ◽  
First Episode ◽  
Genotype 4 ◽  
Dna Concentration ◽  
Healthy Women

Objective. To assess the importance of identifying different genotypes of Gardnerella vaginalis in the diagnosis of recurrent bacterial vaginosis.Materials and methods. The study involved 299 women of reproductive age. All patients were divided into three groups (healthy women, women with the first episode of bacterial vaginosis, and women with recurrent bacterial vaginosis). DNA of Gardnerella vaginalis in vaginal discharge was detected by real-time PCR. The detection of four genotypes of G. vaginalis was performed using real-time multiplex PCR. To quantify the amplified PCR fragments, quantitative standard samples were constructed. Statistical analysis of the results was carried out using the statistical package NCSS 11 (NCSS, LCC).Results. In 38.2 % of healthy women, any one genotype of G. vaginalis was identified in the vaginal biotope, most often it was genotype 4 (35.2 %), while the concentration of G. vaginalis DNA was low (102–103 geqs/ml). When several genotypes of gardnerella were detected simultaneously in healthy women, the DNA concentration did not exceed 104 geqs/ml. A completely different picture was observed among women with bacterial vaginosis (BV). In the first episode of BV, genotype 4 of G. vaginalis prevailed, both as a single genotype and in combination with 1 or 2, or 3 genotypes. In the recurrent course of BV, only 3–4 genotypes of G. vaginalis were detected at once, and in 78 % of cases it had place is a combination of 1, 2 and 4 genotypes, and the DNA concentration was 107–108 geqs/ml.Conclusion. To diagnose recurrent forms of BV, it is necessary to develop and introduce into practice laboratory diagnostics a test system for detecting different genotypes of G. vaginalis by real-time PCR.

scholarly journals THE CLINICAL AND MICROBIOLOGICAL FEATURES OF BACTERIAL VAGINOSIS

2017 ◽  
pp. 15-18
Author(s):  
Yu. A. Lyzikova
Keyword(s):  
Bacterial Vaginosis ◽  
Sexually Transmitted Infections ◽  
Reproductive Function ◽  
Gardnerella Vaginalis ◽  
Control Group ◽  
Sexually Transmitted ◽  
Female Patients ◽  
Microbiological Examination ◽  
Lactobacillus Spp ◽  
Atopobium Vaginae

Aim : to determine the clinical and microbiological features of bacterial vaginosis on the basis of the microbiological examination, assessment of the cytokine status in female patients. Material and methods . The article presents the results of the complex clinical and microbiological examination of 86 female patients of the fertile age. 30 (34.88 ± 5.14 %) patients were diagnosed bacterial vaginosis on the basis of revelation and identification of DNA of Gardnerella vaginalis, Atopobium vaginae, Lactobacillus spp. and the total number of bacteria. The control group consisted of 56 (65.12 ± 5.14 %) patients without bacterial vaginosis. The work also presents the results of the microbiological analysis of the material obtained from the cervical canal and endometrium. All the patients underwent blood tests for detection of the inflammatory reaction - interleukins IL-1, IL-2, tumor necrosis factor (TNF-α), interferon (γ-IFN). Results . The prevalence of bacterial vaginosis among the patients of the reproductive age was 34.8 %. The pathology of the reproductive function was found with equal frequency in the patients of both the groups. Disorders of the immune status in favor of pro-inflammatory cytokines were not diagnosed. The clinical and laboratory criteria made it possible to diagnose bacterial vaginosis in 3.49 % of the patients, the use of PCR diagnosis - in 34.88 %. The concentration of lactobacillus spp. is reliably lower in the patients with bacterial vaginosis, than in the control group (p = 0.0085). As for the concentrations of Gardnerella vaginalis and Atopobium vaginae the groups do not significantly differ. Only 4 (13.33 ± 6.21 %) patients (χ = 5.51, p = 0.02) in the main group detected sexually transmitted infections, which should be taken into account while performing the diagnostic activities. Conclusion. The identification of DNA of certain kinds of microorganisms give an opportunity to assess the state of vaginal microcenosis and the degree of its malfunction even in the absence of clinical and other laboratory signs of bacterial vaginosis. The malfunction of the biocenosis of the genital tract is not associated with disorders of the reproductive function and does not lead to a change in the cytokine status. Patients with bacterial vaginosis are in the risk group for development of sexually transmitted infections, which should be taken into account while performing the diagnostic activities.

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 ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Pregnant Women ◽  
Real Time ◽  
Vaginal Discharge ◽  
Gardnerella Vaginalis ◽  
Bacterial Number ◽  
Vaginal Smears ◽  
Vaginal Lactobacilli ◽  
Real Time Qpcr ◽  
Atopobium Vaginae

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.

scholarly journals Dynamics of Vaginal Bacterial Communities in Women Developing Bacterial Vaginosis, Candidiasis, or No Infection, Analyzed by PCR-Denaturing Gradient Gel Electrophoresis and Real-Time PCR

2007 ◽  
Vol 73 (18) ◽  
pp. 5731-5741 ◽  
Author(s):  
Beatrice Vitali ◽  
Ciro Pugliese ◽  
Elena Biagi ◽  
Marco Candela ◽  
Silvia Turroni ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Real Time ◽  
Relative Abundance ◽  
Real Time Pcr ◽  
Bacterial Communities ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Pcr Analysis ◽  
Healthy Women ◽  
Gradient Gel Electrophoresis

ABSTRACT The microbial flora of the vagina plays a major role in preventing genital infections, including bacterial vaginosis (BV) and candidiasis (CA). An integrated approach based on PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and real-time PCR was used to study the structure and dynamics of bacterial communities in vaginal fluids of healthy women and patients developing BV and CA. Universal eubacterial primers and Lactobacillus genus-specific primers, both targeted at 16S rRNA genes, were used in DGGE and real-time PCR analysis, respectively. The DGGE profiles revealed that the vaginal flora was dominated by Lactobacillus species under healthy conditions, whereas several potentially pathogenic bacteria were present in the flora of women with BV. Lactobacilli were the predominant bacterial population in the vagina for patients affected by CA, but changes in the composition of Lactobacillus species were observed. Real-time PCR analysis allowed the quantitative estimation of variations in lactobacilli associated with BV and CA diseases. A statistically significant decrease in the relative abundance of lactobacilli was found in vaginal fluids of patients with BV compared to the relative abundance of lactobacilli in the vaginal fluids of healthy women and patients with CA.

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