无乳链球菌恒温热隔绝式PCR快速检测方法的建立  

Establishment of insulated isothermal PCR assay of Streptococcus agalactiae

作　　者：王娟 卞国志 贾坤[2] 刁巧虹[2] 李守军 王贵平[1] 袁建丰 

机构地区：[1]广东海大畜牧兽医研究院有限公司,广东广州511400 [2]华南农业大学兽医学院,广东广州510642

出　　处：《中国预防兽医学报》2017年第9期731-733,745,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：广东省科技计划项目(2014A020208052);广州市科技计划项目(201510010250)

摘　　要：为建立快速检测无乳链球菌的方法,本研究针对无乳链球菌表面免疫原性蛋白sip基因的保守区域设计特异性引物,建立了快速检测无乳链球菌的恒温热隔绝式PCR(iiPCR)方法。该检测方法能在30 min内迅速扩增出大量大小为105 bp的目的条带;并与肺炎链球菌、金黄色葡萄球菌、枯草芽孢杆菌、乳房链球菌、停乳链球菌均无交叉反应;检测下限为10~2拷贝/μL,优于普通PCR方法。本研究建立的检测方法具有操作简便、高特异性、高敏感性及高扩增效率的特点,适用临床或基层实验室使用。Insulated isothermal PCR （iiPCR）, established on the basis of Raylcigh-Benard convection, is a rapid and low-lost platform for nucleic acid amplifiation. A rapid insulated isothermal PCR （iiPCR） assay was established to detect Streptococcus agalactiae based on sip gene in this study. The method could rapidly amplify a large number of 105 bp target fragments within 30 minutes. The detection results showed that the iiPCR assay was specific for S. agalactiae detection with a detection limit of 102 copies/μL, but no cross-reactions with other pathogenic bacteria were found. In conclusion, the iiPCR method is a simple, rapid and sensitive technique for detection of S.agalactiae in the field or laboratory.

关 键 词：无乳链球菌 恒温热隔绝PCR 表面免疫原性蛋白SIP 

分 类 号：S852.61[农业科学—基础兽医学]