布鲁菌病半套式PCR检测方法的建立  

Establishment of Rapid Test of Brucella by Nest-PCR

作　　者：马鸣潇[1] 刘锴[2] 于洋[1] 周铁忠[1] 张利博[1] 

机构地区：[1]锦州医学院畜牧兽医学院,辽宁锦州121001 [2]内蒙古民族大学动科院,内蒙古通辽028000

出　　处：《锦州医学院学报》2006年第5期5-6,17,共3页Journal of Jinzhou Medical College

基　　金：辽宁省教育厅资助项目(编号:20040130)

摘　　要：目的建立布鲁菌属半套式PCR检测方法。方法根据布鲁菌BCSP-31蛋白基因设计三条引物,扩增目的片段长223 bP,通过条件优化,建立了布鲁菌属半套式PCR检测方法。利用所建立的PCR方法对六个种布鲁菌和Y.Center O:9,S.TyPh i,E.Coli O:157进行检测验证其特异性,通过对活菌计数检测验证其敏感性。结果检测结果表明六个种布鲁菌均可扩出223bP的目的片段,而Y.Center O:9,S.TyPh i和E.Coli O:157不能扩出目的片段。通过对活菌计数最低可检测到20个细菌。结论本研究建立了敏感、特异的布鲁菌病半套式PCR快速检测方法,为布鲁菌的检测和试剂盒的研制奠定了基础。Objective To establish nest - PCR method of Brucella . Methods According to B. abortus 31KD gene , a set of primers to detect Brucella were designed. The Production of the PCR assay is 223 bp. The optimal condition of PCR for detection of Brucella was determined through orthogonal assay. The specificity of the sets of primers were examined by PCR using template extracted from of 6 species Brucella , Y. enterocolitica O： 9, E. coli 157 and S. tyPhimuium strains. While the sensitivity of PCR was also determined by a seral dilution of Brucella. Results The DNA of 6 species Brucella strains were amplified by the primers of 31 KDa, but not for the DNA of Y. enterocolifica O ： 9, E. coli 157 and S. typhimuium strains, about 20 CFU can be detected. This shows that the nest - PCR method of Brucella is specificity and sensitivity. Conclusions Nest - PCR method of Brucella has been established in this study , and veritified to be specificity and sensitivity. The study has paved the way for testing of Brucella.

关 键 词：布鲁菌 半套式PCR 检测 

分 类 号：Q939[生物学—微生物学]