摘要
为建立快速有效、经济适用的沙门氏菌血清型鉴定方法,利用Primer Plex软件设计畜禽中常见沙门氏菌的菌体(O)抗原和鞭毛抗原(H)的引物,优化引物比例和退火温度等反应条件,建立沙门氏菌血清型鉴定的PCR法,并对46株沙门氏菌分离株进行血清型鉴定,同时与传统玻片凝集法和美国CDC的液相芯片(Luminex-x MAP)法进行比较。优化后获得了O抗原中B、C1、D和E1群的多重PCR引物,以及鞭毛抗原H1相抗原基因(fli C)和H2相抗原基因(flj B)的2对引物;建立了一套基于沙门氏菌O和H抗原的多重PCR血清型鉴定法;PCR法和Luminex-x MAP法鉴定出7种沙门氏菌血清型,传统玻片凝集法鉴定出6种血清型;PCR法与传统玻片凝集法和Luminex-x MAP鉴定结果的符合率均为95.65%。本研究建立的PCR沙门氏菌血清型鉴定法简便、快速,预期可在沙门氏菌的血清分型研究中发挥作用。
In order to develop a rapid,effective,economic and applicable method for identifying Salmonella serotypes,the primers for O antigen and flagellum antigen（H）of common Salmonella bacteria in livestock and poultry were designed,and PCR method was established by optimizing the concentration of primers and the anneal temperature. Forty-six Salmonella isolates were tested by the PCR method,and the results of which were compared to the traditional agglutination method and the Luminex-xMAP method. After optimization,multi-primers of group B,C1,D,E1 in antigen O were obtained,as well as the antigens H1 and H2 encoded by genes of fiC and fjB. Then a set of multiple PCR method was established on the basis of Salmonella O and H antigens. Results showed 7 serotypes were identi？ed by this method and Luminex-xMAP method,and 6 serotypes were identi？ed by traditional agglutination test. The coincidence rate between traditional agglutination test and Luminex-xMAP and the PCR method was 96.56 %. The PCR assay was simple and rapid,and it would play important role in Salmonella serotype evaluation.
作者
赵建梅
李月华
宋传周
赵格
曲志娜
Zhao Jianmei;Li Yuehua;Song Chuanzhou;Zhao Ge;Qu Zhina(China Animal Health and Epidemiology Center,Qingdao,Shandong266032,China;Wangmiao Animal Inspection Branch,Pingyuan Animal Husbandry and Veterinary Bureau,Dezhou,Shandong253100,China)
出处
《中国动物检疫》
CAS
2018年第1期73-77,共5页
China Animal Health Inspection
基金
国家"十二五"科技基础性工作专项(2012FY111000)
关键词
沙门氏菌
血清型
PCR
鉴定
符合率
Salmonella
serotype
PCR
identification
coincidence rate
