Longitudinal Analysis Of Vaginal Microbiota In Women With Recurrent Vulvovaginal Candidiasis

BackgroundRecurrent vulvovaginal candidiasis(RVVC),refers to after treatment,the clinical symptoms disappeared and mycological examination negative,recurrence 4 episodes or more within 1 years.At present,local antifungal therapy is the main treatment for RVVC,but the recurrence of the disease is the bottleneck of disease treatment.In recent years,it has been emphasized that the change of vagina flora plays an important role in the occurrence and development of RVVC.But at present,there are some controversies about the research results of vaginal flora of RVVC.This study intends to collect the self-swabs of vaginal flora at different time in patients with RVVC and health women and then by Illumina high-throughput sequencing technology and use the BIPES biological information analysis method to analysis the vaginal flora structure and dynamic change between RVVC group and control group.Our study maybe can provide a theoretical basis for the study of the mechanism of RVVC recurrence and guiding clinical treatment.ObjectivesTo explore the dynamic change of vaginal flora in patients with recurrent vulvovaginal candidiasis(RVVC).Methods1.Experimental object:A total of 6 RVVC patients who went to the department of obstetrics and gynecology of Zhujiang Hospital of Southern Medical University from December 2015 to June 2016 were recruited in this study,and a total of 5 healthy reproductive age women were recruited as control group.2.Samples collection:After informed the experimental object of the experiment methods and signed informed consent,asked them use self-swabs to rotate in the lower segment of vagina 1/3 at least 10 times at least every 3 days at least 2 months,and keep swabs in-20 low temperature refrigerator first,then transferred swabs to the-80 low temperature refrigerator at regular intervals.3.Experimental methods:Genomic DNA was extracted by genomic DNA purification kit of each individual.We used extracted genome DNA as a template,amplified the 16S rRNA V4 gene fragment,and the PCR products were mixed with equal final concentration,and then sequenced by platform of Illumina Hiseq 2000.Through BIPES biological information analysis method to analysis and comparison of the structure and dynamics of the vaginal flora of the two groups.4.Statistical treatment:We used the SPSS20.0 software for statistical analysis.The Mann-Whitney U test was used to compare the diversity index and the relative abundance of each individual.P<0.05 was found to be statistically significant.Results:1.Except for Lactobacillus,Gardnerella and Prevotella can also be predominant species for healthy women,When Lactobacillus is the predominant species,the majority of Lactobacillus were L.crispatus.Compared with the control group,the diversity of vaginal flora in RVVC patients was significantly reduced and L.iners was the predominant species in most RVVC patients.Content of L.crispatus decreased significantly in RVVC patients.2.Fluctuate with changes in vaginal flora of the menstrual cycle in healthy women,and during menstruation vaginal flora fluctuate most obvious.Presented alternately dominant flora or change mode to maintain absolute superiority in the menstrual cycle.The vaginal flora of RWC patients was relatively stable,the volatility of vaginal flora weakened in RVVC patients and the structure of the flora was similar in the menstrual period,attack period and intermittent period.Conclusion:1.The vaginal flora of RWC patients was relatively stable and the structure of the flora was similar in the menstrual period,attack period and intermittent period;2.The L.iners were the predominant species in most RVVC patients;3.The treatment of RWC should pay attention to the regulation of vaginal micro-environment,induce the production of L.iners or inhibit the colonization of L.crispatus to recovery the proportion of vaginal flora may be could cure RWC.