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

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(deleted region 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H37Rv和H37Ra标准菌株、48株从结核病患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。结论子多重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。

Application of multiplex polymerase chain reaction in rapid identification of Mycobacterium bovis BCG

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.