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    荧光PCR法在围生期孕妇B族链球菌筛查中的应用及CAMP试验阴性B族链球菌分子特征分析

    Application of fluorescent PCR in screening of group B streptococcus in perinatal pregnant women and analysis of molecular characteristics of group B streptococcus with negative CAMP test

      摘要
    • 摘要:
      目的探讨荧光聚合酶链反应(PCR)在围生期孕妇B族链球菌(GBS)筛查中的应用价值及CAMP试验阴性GBS的鉴定分析。
      方法收集产科门诊孕35~37周孕妇阴道/直肠拭子1 143例，将拭子放入GBS增菌肉汤管增菌，分别用GBS显色平板和荧光PCR法检测GBS。16SrDNA分子鉴定技术对CAMP试验阴性GBS进行种属鉴定，并采用PCR技术检测其cfb基因。
      结果显色平板检出146份阳性样本，阳性率为12.77%，荧光PCR法检出191份阳性样本，阳性率为16.71%，2种方法的阳性率差异有统计学意义(P<0.01)；荧光PCR法与显色平板培养法相比，敏感性为93.15%、特异性为94.48%、符合率为94.31%。146株GBS中有10株为CAMP试验阴性，检出率为6.85%，均为cfb基因缺失株。
      结论荧光PCR法可有效提高围生期孕妇GBS筛查的阳性率；该地区CAMP试验阴性GBS检出率较高，CAMP试验不宜作为GBS的鉴别试验，因存在缺失cfb基因GBS，GBS分子检测试剂不应选择该段基因设计引物。

       

      Abstract:
      ObjectiveTo explore the application value of fluorescent polymerase chain reaction (PCR) in screening of group B streptococcus (GBS) in perinatal pregnant women and the identification and analysis of GBS with negative CAMP test.
      MethodsA total of 1 143 vaginal/rectal swabs were collected from pregnant women aged 35-37 weeks in the obstetric clinic.The swabs were placed in a GBS enrichment broth tube to enrich the bacteria, and then GBS chromogenic plate and fluorescent PCR were used to detect GBS.16SrDNA molecular identification technology was used to identify the species of GBS with negative CAMP test, and PCR was used to detect its cfb gene.
      ResultsThe chromogenic plate detected 146 positive samples, with a positive rate of 12.77%, and the fluorescent PCR detected 191 positive samples, with a positive rate of 16.71%.The difference between the two methods was statistically significant (P<0.01).Compared with the chromogenic plate culture method, the fluorescent PCR method had a sensitivity of 93.15%, a specificity of 94.48%, and a coincidence rate of 94.31%.Among the 146 GBS strains, 10 strains were negative for CAMP test, with a detection rate of 6.85%, and all strains were cfb gene deletion.
      ConclusionsThe fluorescent PCR can effectively increase the positive rate of GBS screening for pregnant women during the perinatal period.The detection rate of GBS with negative CAMP test is relatively high in this area, and CAMP test should not be used as an identification test for GBS.Due to the deletion of cfb gene, GBS molecular detection reagents should not select this gene to design primers.

       

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