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目的:确定荧光定量PCR方法检测细菌性阴道病相关细菌阴道加特纳菌的可行性。方法:采用SYBR Green方法建立了阴道加特纳菌荧光定量PCR方法,评价荧光PCR方法的特异性、灵敏度、重复性;并对568例临床分泌物样本进行检测。结果:本实验所建立的荧光定量PCR方法可准确、特异的检测阴道加特纳菌;该方法的灵敏度可达到1拷贝/μl;194例细菌性阴道病患者分泌物标本中荧光定量PCR法检出116例(59.79%)阴道加特纳菌;而在374例非细菌性阴道病患者分泌物标本中荧光定量PCR法检出35例(9.36%)阴道加特纳菌;荧光定量PCR法检出率明显高于细菌培养方法,差异具有显著性,χ2=24.89,P<0.01。结论:荧光定量PCR方法能够应该用于临床分泌物标本中的阴道加特纳菌的检测。
OBJECTIVE: To determine the feasibility of fluorescence quantitative PCR for the detection of Gardnerella vaginalis bacteria associated with bacterial vaginosis. Methods: SYBR Green method was used to establish the Gardnerella vaginalis fluorescence quantitative PCR method to evaluate the specificity, sensitivity and repeatability of fluorescence PCR. 568 clinical secretions samples were also tested. Results: The fluorescence quantitative PCR method established in this experiment can accurately and specifically detect Gardnerella vaginalis; the sensitivity of this method can reach 1 copy / μl; 194 cases of bacterial vaginosis secretions specimens by fluorescence quantitative PCR method 116 cases (59.79%) were found to have Gardnerella vaginalis, while 35 cases (9.36%) of Gardnerella vaginalis were detected by fluorescence quantitative PCR in 374 cases of non-bacterial vaginosis. Fluorescent quantitative PCR Rate was significantly higher than the bacterial culture method, the difference was significant, χ2 = 24.89, P <0.01. Conclusion: The fluorescence quantitative PCR method should be applied to the detection of Gardnerella vaginalis in clinical specimens.