基于实时定量PCR技术的育龄期细菌性阴道病患者阴道菌群结构分析及其临床应用

Structural analysis and clinical application of vaginal microbiota in patients with bacterial vaginosis in reproductive-age women based on real-time quantitative PCR

目的探讨育龄期女性细菌性阴道病(bacterial vaginosis,BV)患者阴道优势菌群变化及其与临床指标的相关性。方法选择本院门诊确诊的育龄期BV患者35例和同期入院体检的健康体检女性37例,无菌拭子采集阴道中段壁分泌物,提取细菌基因组DNA,采用实时定量PCR技术(real-time quantitative PCR,qPCR)进行阴道优势细菌检测,并进行其与临床指标如阴道pH和Nugent评分相关性分析。结果乳杆菌属细菌及其种水平的细菌如惰性乳杆菌、卷曲乳杆菌、詹氏乳杆菌等在BV患者中均显著下降,BV相关阴道致病细菌如加德纳菌属、奇异菌属、埃格特菌属、巨型球菌I型菌属、纤毛菌属和普氏菌属显著升高(均P<0.05)。阴道致病菌群与阴道pH和Nugent评分呈显著正相关,而阴道卷曲乳杆菌和惰性乳杆菌与其呈负相关。结论育龄期BV患者阴道优势菌群显著失衡,并与阴道pH和Nugent评分显著相关,提示阴道优势菌群改变参与BV发生发展。

Objective To investigate the changes of vaginal predominant bacteria and its correlations with clinical indicators in patients with bacterial vaginosis(BV)in reproductive-age women.Methods Thirty-five BV patients and thirty-seven healthy(CN)women,who presented to the department of obstetrics and gynecology of our hospital,were recruited for this study.When women underwent genital examinations,samples were obtained near the mid-vagina using a sterile swab,packaged,and placed in ice packs.Then bacterial DNA was extracted from the vaginal swabs using a QIAamp DNA Mini Kit(QIAGEN,Hilden,Germany)according to the manufacturer’s instructions.With real-time quantitative PCR(qPCR),we investigated the loads and relative abundance of the vaginal predominant bacteria,and its correlations between vaginal predominant bacteria and clinical indicators such as vaginal pH and Nugent score.Results Our present findings showed a significant decline in Lactobacillus genus,and its species,such as L.iners,L.crispatus and L.jensenii in BV patients when compared with healthy controls,while vaginal pathogenic bacteria such as Gardnerella vaginalis,Atopobium vaginae,Leptotrichia/Sneathia spp.,Eggerthella spp.,Megasphaera typeⅠspp.and Prevotella spp.were significantly increased in BV patients(P<0.05).In addition,the vaginal pathogenic bacteria were correlated positively with vaginal pH and Nugent scores,while Lactobacillus spp.,such as L.iners and L.crispatus were negatively correlated with them(P<0.05).Conclusion Vaginal dysbiosis was found in BV patients in reproductive-age women.The vaginal predominant bacteria were significantly correlated with vaginal pH and Nugent score,suggesting that vaginal dominant bacteria played vital roles in BV pathogenesis.