scholarly journals Complexity of Vaginal Microflora as Analyzed by PCR Denaturing Gradient Gel Electrophoresis in a Patient With Recurrent Bacterial Vaginosis

2005 ◽  
Vol 13 (1) ◽  
pp. 25-30 ◽  
Author(s):  
Estelle Devillard ◽  
Jeremy P. Burton ◽  
Gregor Reid

Objective:Gardnerella vaginalishas long been the most common pathogen associated with bacterial vaginosis (BV). We aimed to test our hypothesis that symptoms and signs of BV do not necessarily indicate colonization by this organism, and often will not respond to standard metronidazole or clindamycin treatment.Methods:Using a relatively new molecular tool, PCR denaturing gradient gel electrophoresis (DGGE), the vaginal microflora of a woman with recalcitrant signs and symptoms of BV was investigated over a 6-week timeframe.Results:The vagina was colonized by pathogenic enterobacteriaceae, staphylococci andCandida albicans. The detection of the yeast by PCR-DGGE is particularly novel and enhances the ability of this tool to examine the true nature of the vaginal microflora. The patient had not responded to antifungal treatment, antibiotic therapy targeted at anaerobic Gram-negative pathogens such asGardnerella, nor daily oral probiotic intake ofLactobacillus rhamnosusGG. The failure to find the GG strain in the vagina indicated it did not reach the site, and the low counts of lactobacilli demonstrated that therapy with this probiotic did not appear to influence the vaginal flora.Conclusions:BV is not well understood in terms of its causative organisms, and further studies appear warranted using non-culture, molecular methods. Only when the identities of infecting organisms are confirmed can effective therapy be devized. Such therapy may include the use of probiotic lactobacilli, but only using strains which confer a benefit on the vagina of pre- and postmenopausal women.

2018 ◽  
Vol 12 (02) ◽  
pp. 127-136 ◽  
Author(s):  
Laura Maria Andrade Oliveira ◽  
Cláudio Galuppo Diniz ◽  
Aline Augusta Sampaio Fernandes ◽  
Daniele Maria Knupp Souza-Sotte ◽  
Michelle Cristine Ribeiro Freitas ◽  
...  

Introduction: Bacterial vaginosis (BV) is characterized by the depletion of Lactobacillus spp. population and increase of other species, especially Gardnerella vaginalis and Atopobium vaginae. This study aimed to investigate the vaginal microbiota structure of Brazilian women with and without BV according to Nugent Score and to assess the correlation among Nugent score and the quantification of BV-associated bacteria. Methodology: Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) assay was employed to characterize the vaginal microbiota structure. Quantification of Lactobacillus spp., G. vaginalis, A. vaginae, Mobiluncus sp. and M. hominis were determined by quantitative real-time PCR (qPCR). Results: Clustering by PCR-DGGE revealed differences in microbial structure of the different patient groups. Gardnerella vaginalis, A. vaginae, M. hominis and Mobiluncus sp. were detected at high loads in BV-associated microbiota. Quantification of Lactobacillus spp. showed an inverse correlation with Nugent score while the loads of G. vaginalis, A. vaginae, M. hominis and Mobiluncus sp. indicated a direct correlation with this method. Conclusions: Despite Nugent score is considered the gold standard for BV diagnosis, qPCR stands out as a useful tool for bacteria quantification and an alternative for BV diagnosis. Vaginal microbiota is a complex microbial community although there is a common core among BV and non-BV women. Investigation of vaginal microbiota structure may contribute to the development of tools for diagnosis improvement and therapeutic regimen optimization.


2008 ◽  
Vol 74 (14) ◽  
pp. 4539-4542 ◽  
Author(s):  
Rafael C. R. Martinez ◽  
Sílvio A. Franceschini ◽  
Maristela C. Patta ◽  
Silvana M. Quintana ◽  
Álvaro C. Nunes ◽  
...  

ABSTRACT Culture-dependent PCR-amplified rRNA gene restriction analysis and culture-independent (PCR-denaturing gradient gel electrophoresis) methodologies were used to examine vaginal lactobacilli from Brazilian women who were healthy or had been diagnosed with vulvovaginal candidiasis (VVC) or bacterial vaginosis. Only Lactobacillus crispatus was detected accordingly by both methods, and H2O2-producing lactobacilli were not associated with protection against VVC.


2007 ◽  
Vol 73 (18) ◽  
pp. 5731-5741 ◽  
Author(s):  
Beatrice Vitali ◽  
Ciro Pugliese ◽  
Elena Biagi ◽  
Marco Candela ◽  
Silvia Turroni ◽  
...  

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.


2020 ◽  
Vol 51 (2) ◽  
pp. 125-146
Author(s):  
Nasiruddin Nasiruddin ◽  
Yu Zhangxin ◽  
Ting Zhao Chen Guangying ◽  
Minghui Ji

We grew cucumber in pots in greenhouse for 9-successive cropping cycles and analyzed the rhizosphere Pseudomonas spp. community structure and abundance by PCR-denaturing gradient gel electrophoresis and quantitative PCR. Results showed that continuous monocropping changed the cucumber rhizosphere Pseudomonas spp. community. The number of DGGE bands, Shannon-Wiener index and Evenness index decreased during the 3rd cropping and thereafter, increased up to the 7th cropping, however, however, afterwards they decreased again. The abundance of Pseudomonas spp. increased up to the 5th successive cropping and then decreased gradually. These findings indicated that the structure and abundance of Pseudomonas spp. community changed with long-term cucumber monocropping, which might be linked to soil sickness caused by its continuous monocropping.


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