B族链球菌恒温荧光探针检测方法的建立及方法学验证  

The Development and Validation of a Rapid Molecular DetectionSystem with Fluorescence Probe Isothermal Amplificationfor Group B Streptococcus

作　　者：许朝霞 张省委 丁程伟 张慧芸 相星 XU Zhaoxia;ZHANG Shengwei;DING Chengwei;ZHANG Huiyun;XIANG Xing(Department of Clinical Laboratory,East Hospital of Beijing University of Chinese Medicine,Beijing 100078,China)

机构地区：[1]北京中医药大学东方医院检验科,北京100078

出　　处：《标记免疫分析与临床》2023年第9期1560-1565,共6页Labeled Immunoassays and Clinical Medicine

基　　金：丰台区卫生健康系统科研课题(编号:2020-80)。

摘　　要：目的 建立B族链球菌恒温荧光探针检测方法体系。方法 针对B族链球菌高度保守区域设计特异性引物和含RNA碱基的探针,建立B族链球菌酶切探针恒温扩增快速分子检测体系,并从检测限、灵敏度、特异性和抗干扰能力、精密度、准确性进行方法学验证。结果 该方法的检测限为102CFU/mL,灵敏度为100.0%,能检测出B族链球菌的5种亚型(Ia、Ib、II、III、V型),对于其他细菌的检测均为阴性,显示该方法特异性良好。检测108份阴道拭子的临床标本,检测结果与临床培养结果相一致,准确性100.0%。精密度检测显示变异系数均小于5.0%,精密度良好。结论 该方法的灵敏度高,特异性和精密度良好,操作简便、高效快捷,30min即可扩增完成,可用于B族链球菌的快速检测。Objective To establish a rapid molecular detection system of group B Streptococcus with enzymatic probe isothermal amplification.Methods We designed specific primers and probes containing RNA bases for highly conserved regions of group B Streptococcus and established a rapid molecular detection system for isothermal amplification of group B Streptococcus with enzymatic probe.The system was then validated by sensitivity and detection limit,specificity and anti-interference ability,repeatability and precision,and accuracy.Results The detection limit of this system was 102 CFU/mL,with sensitivity of 100.0%.This method was able to detect five subtypes of group B Streptococcus(Ia,Ib,II,III and V),while no other bacteria could be detected,indicating that the specificity of this method was good.108 vaginal swabs were tested by this method subsequently and results were consistent with the clinical culture results.The precision test showed that the coefficient of variation was less than 5.0%,indicating a good precision.Conclusion The developed method in this study has a high sensitivity,good specificity and precision.In addition,the easy operation and high efficiency make it a great choice for the rapid detection of group B Streptococcus.

关 键 词：B族链球菌 酶切探针 阴道拭子 恒温扩增 验证 

分 类 号：R378.12[医药卫生—病原生物学] R-331[医药卫生—基础医学]