TaqMan荧光探针技术检测不同血清型B群链球菌的方法建立及评价  

Establishment and evaluation of different serotypes group B streptococcus method by TaqMan fluorescent probe technique

作　　者：江凡[1] 王飞玲[1] 刘小月[1] 路瑞静 周喜友[1] 邹建话[1] JIANG Fan;WANG Feiling;LIU Xiaoyue;LU Ruijing;ZHOU Xiyou;Zou Jianhua(Department of Clinical Laboratory,Maternity and Child Health Care Institute of Baoan District,Shenzhen,Guangdong 518100,China)

机构地区：[1]深圳市宝安区妇幼保健院检验科,广东深圳518100

出　　处：《国际检验医学杂志》2018年第4期438-442,共5页International Journal of Laboratory Medicine

基　　金：深圳市知识创新计划项目(JCYJ20150403105513694)

摘　　要：目的建立基于TaqMan荧光探针技术,对不同血清型B群链球菌(GBS)鉴定的分子方法,为后续研究多重荧光探针技术检测不同血清型GBS奠定基础。方法根据不同血清型荚膜多糖(CPS)序列的差异设计引物和探针,用实时荧光定量PCR扩增CPS序列,建立不同血清型的GBS分型方法,从灵敏度、特异度及临床分离株检测等方面对该方法进行评价,并与胶乳凝集试验结果比较。结果同一血清型GBS的DNA的对数浓度与循环阈值(Ct)呈良好的线性相关,该方法的检测下限均为1pg/μL,一种探针只能检测对应血清型的GBS。10株标准菌株的TaqMan荧光探针技术检测结果与胶乳凝集试验结果一致。结论 TaqMan荧光探针技术是一种简单、快速、具有较高灵敏度和特异度的检测GBS血清型的方法,对临床分离株的分型优于乳胶凝集试验。Objective To establish a molecular method for the identification of different serotypes of group B streptococcus(GBS)based on TaqMan fluorescence probe technology,and to lay the foundation for the subsequent study of multiple fluorescent probe technology to detect different serotypes of GBS.Methods Primers and probes were designed according to the different serotypes of capsular polysaccharide(CPS).CPS sequences were amplified by real-time fluorescence quantitative polymerase chain reaction.GBS classification methods of different serotypes were established.The results were compared with latex agglutination test and the method was evaluated from the aspects of sensitivity,specificity and detection of clinical isolates.Results The logarithmic concentration of DNA in the same serotype GBS was linearly correlated with the value of Ct.The detection limit of this method is 1 pg/μL,a probe could only detect the corresponding serotype GBS.The results of TaqMan fluorescence probe test of 10 strains were consistent with the results of latex agglutination test.Conclusion TaqMan fluorescence probe technique is a simple,rapid,highly sensitive and specific method for the detection of different GBS serotypes,and it is better than latex agglutination test for the classification of clinical isolates.

关 键 词：TaqMan荧光探针 实时荧光定量聚合酶链反应 B群链球菌 

分 类 号：R446.5[医药卫生—诊断学]