细菌16SrRNA基因PCR诊断细菌性阴道病的研究

Diagnosis of bacterial vaginosis by 16SrRNA genes PCR

目的确定阴道主要角色菌的分离率与细菌性阴道病(bacterial vaginosis,BV)的关系,建立BV的分子诊断方法。方法采用临床Amsel标准筛选91例BV患者、82例健康对照妇女。采集阴道分泌物标本提取细菌基因组DNA,采用细菌特异性16SrRNA基因PCR检测标本中的卷曲乳酸杆菌、阴道加德纳菌、人型支原体等11种角色菌。比较上述细菌分离率与BV发生的关系。通过统计学分析建立最佳的用于BV诊断细菌检测组合。结果与BV密切相关的阴道细菌有纤毛菌属、阴道阿托波菌属、巨球菌属、羞怯动弯杆菌属、普雷沃菌属。上述细菌的分离率在2组样本的差异具有统计学意义(P<0.01)。以联合检测阴道阿托波菌属、纤毛菌属、普雷沃菌属作为BV的判断标准时,敏感性为93.41%,特异性为78.05%。结论 BV为多种微生物混合感染性疾病,通过PCR方法检测主要致病菌或致病菌组合,可以有效地对BV进行快速分子诊断。

Objective To determine the organism＇s isolation rate in bacterial vaginosis（BV）,to establish a molecular diagnostic method for BV.Methods Totally 173 vaginal-fluid samples（91 with BV and 82 normal） were screened by clinical Amsel criteria.Bacterial genome DNA was extracted from the vaginal secretion samples.Eleven major vaginal bacterial species were detected by using 16SrRNA gene PCR assays to confirm its association with BV,and the utility of PCR for the microbiological diagnosis of BV.Statistical analysis was used to establish the best bacterial combination for diagnosis of BV.Results Leptotrichia/Sneathia,Atopobium vaginae,Megasphaera species,Mobiluncus mulieris and Prevotella among the bacterial species were significantly associated with BV,the organism＇s isolation rate between two groups had significant difference（P0.01）.As a diagnostic criteria of BV,getting a sensitivity of 93.41% and a specificity of 78.05% by co-detecting the three species,Atopobium vaginae,Leptotrichia and Megasphaera.Conclusion BV is a polymicrobial disease.PCR detection of one or more fastidious bacterial species is an effective reliable molecular diagnostic method.