应用多重聚合酶链反应技术快速鉴定牛分支杆菌卡介苗株  

Application of multiplex polymerase chain reaction in rapid identific ation of Mycobacterium bovis BCG

作　　者：李国利[1] 陈保文[2] 庄玉辉[1] 王国治[2] 赵铭[1] 沈小兵[2] 孙桂芝[3] 

机构地区：[1]解放军第三○九医院结核病研究室,北京100091 [2]中国药品生物制品检定所 [3]北京市胸科医院

出　　处：《中华结核和呼吸杂志》2003年第11期701-703,共3页Chinese Journal of Tuberculosis and Respiratory Diseases

摘　　要：目的　建立快速鉴定牛分支杆菌卡介苗 (BCG)菌株的方法。方法　依据最近研究发现的牛分支杆菌BCG菌株不存在命名为缺失区 1(deletedregion 1,RD1)的基因区 ,而其他牛分支杆菌菌株和其他结核分支杆菌复合群 (MTC)菌种 (结核分支杆菌、非洲分支杆菌和田鼠分支杆菌 )存在RD1基因区的遗传学信息 ,应用多重聚合酶链反应 (PCR)检测RD1基因区存在与否 ,以鉴别BCG菌株与其他牛分支杆菌菌株及其他MTC菌种。结果　所试 5株BCG疫苗生产用标准菌株和近期国内发生的 1例小儿BCG接种后全身播散性感染致死病例的 2株BCG分离菌株 ,均缺失RD1基因区 ;其他牛分支杆菌菌株 ,包括 3株牛分支杆菌标准菌株、5株分别从结核病牛或鹿分离的牛分支杆菌菌株 ,1株从结核病患者痰标本分离的牛分支杆菌菌株 ,以及其他MTC菌种 ,包括结核分支杆菌H3 7Rv和H3 7Ra标准菌株、48株从结核病患者痰标本分离的结核分支杆菌菌株、3株非洲分支杆菌标准菌株 ,均存在RD1区。结论　多重PCR技术用于牛分支杆菌BCG菌株的鉴定简便、快速、特异 ,适于在临床实验室应用。ObjectiveTo establish a method for the rapid identification of Mycobacterium bovis BCG. Methods A genomic region designated RD1 was found to be deleted from BCG strains, but present in other strains of Mycobacterium bovis and other members of the Mycobacterium tuberculosis complex (MTC) including Mycobacterium tuberculosis, Mycobacterium africanum,and Mycobacterium microti. With this information, a multiplex PCR method, developed to detect the deletion of RD1, was used to differentiate BCG strains from other strains of Mycobacterium bovis and other members of MTC. Results RD1 was shown to be absent in 5 BCG vaccine strains and 2 BCG strains isolated from an infant who died of systemic disseminated infection induced by BCG vaccination, but it was present in 3 Mycobacterium bovis standard strains, 6 Mycobacterium bovis strains isolated from diseased cows, deer or patients with pulmonary tuberculosis and other MTC strains including Mycobacterium tuberculosis H 37Rv and H 37Ra strains, 48 Mycobacterium tuberculosis strains isolated from patients with pulmonary tuberculosis, and 3 Mycobacterium africanum standard strains. Conclusion The multiplex PCR method is simple, rapid, and specific for the identification of BCG among strains of MTC, and is applicable in clinical laboratories.

关 键 词：结核分支杆菌 RD BCG 多重聚合酶链反应 卡介苗 结核病 菌株 结论 缺失 技术 

分 类 号：R446.5[医药卫生—诊断学]