复发性外阴阴道假丝酵母菌病患者阴道微生物的物种组成及其动态变化

Longitudinal analysis of vaginal microbiota in women with recurrent vulvovaginal candidiasis

目的探究复发性外阴阴道假丝酵母菌病(RVVC)患者阴道菌群的动态变迁。方法不同时间点采集RVVC患者(6例)以及正常对照女性(5例)阴道分泌物,分别对每个样本进行细菌总基因组DNA提取、16S r RNA V4区基因扩增及采用Illumina高通量测序技术对扩增的PCR产物进行测序等步骤,然后通过BIPES、TSC、GAST等程序,分析比较两组阴道菌群物种丰度和结构以及菌群动态变化。结果正常对照女性阴道菌群以乳酸杆菌为主,其中惰性乳杆菌和卷曲乳杆菌比例相当,同时存在少量加德纳菌属、普氏菌属和其它菌属;或者以加德纳菌属、普氏菌属、奇异菌属、纤毛菌属等为主要菌属;而RVVC患者阴道菌群丰富度明显降低,以惰性乳杆菌为优势菌属,卷曲乳杆菌含量明显减少。正常对照女性阴道菌群随月经周期的变化出现波动,其中在月经期的波动最为明显,其优势菌属在月经周期中呈现相互交替或保持绝对优势的变化模式;而RVVC患者阴道菌群的波动性减弱,不随月经周期的变化而变化,且在疾病发作期和间歇期的菌群结构相似。结论 RVVC患者阴道菌群结构不随月经周期发生变化,在疾病发作期和间歇期相似。诱导卷曲乳杆菌产生或抑制惰性乳杆菌定植,从而恢复阴道菌群比例,可能可以治愈RVVC。

Objective To investigate the vaginal flora in patients with recurrent vulvovaginal candidiasis （RWC）. Methods Vaginal swabs were collected at different time points from 6 RVVC patients and 5 healthy women of child-bearing age. The dynamic changes, microbiota composition, alpha diversity and beta diversity in the two groups were assessed by analyzing the 16S rRNA V4 hypervariable region amplified from the total genomic DNA from the swabs. Results Lactobacillus was the predominant species in healthy women with similar proportions of Ldners and L.crispatus; small proportions of Gardnerella, Prevotella and other genus were also detected. In some healthy women, the vaginal flora showed a high relative abundance of anaerobic bacteria such as Gardnerella, Prevotella, Atopobium, Sneathia. Compared with the healthy women, patients with RVVC showed a significantly reduced diversity of vaginal flora, where Ldners was the predominant species and the content of L. crispatus decreased significantly. In healthy women, the vaginal flora fluctuated with the menstrual cycle, and the fluctuation was the most prominent during menstruation; the dominant species either alternated regularly or maintain an absolute superiority in the menstrual cycle. The vaginal flora showed attenuated fluctuation in women with RWC, were highly conserved within the menstrual cycle, and maintained a similar composition in the episodes and intermittent periods. Conclusion The vaginal flora of RVVC patients do not undergo regular variations with the menstrual cycle and shows a similar composition between the episodes and intermittent periods. Promoting the production of L.iners or inhibiting the colonization of L.crispatus to restore the composition of the vaginal flora may help in the treatment of RVVC.