多重PCR方法特异性鉴定卡介苗菌株多糖核酸的初探

与结核分枝杆菌H37Rv菌株进行比较,BCG菌株可找到一个特殊的缺失片段RD1,它存在于所有有毒分枝杆菌中,而在所有的卡介苗菌株中均缺失。应用多重PCR方法检测RD1区的存在与否,可以区别BCG和其它有毒的分枝杆菌。卡介菌多糖核酸来源于卡介菌,检测成品中DNA是否含有RD1区,能特异性地鉴别该制品。结果显示牛分枝杆菌标准株和结核分枝杆菌H37Rv存在RD1区;而卡介菌多糖核酸注射液和国内皮内注射用BCG疫苗生产用菌株扩增产物一致,提示均缺失RD1区。因此,这种多重PCR方法适用于卡介菌多糖核酸注射液的特异性鉴别试验。

Preliminary study on the specific identification of polysaccharide nucleic acid of Bacillus Calmette-Guerin using multiplex PCR

Compared with Mycobacterium tuberculosis H37Rv,a special region of difference named RD1 can be founded deleted from BCG,it almost present in all virulent mycobacterium but lost from all BCG strains.Using a multiplex PCR method to detect the presence or absence of RD1 with different products,BCG strains can be specially differentiated from other virulent mycobacterium.For the polysaccharide nucleic acid fraction of Bacillus Calmette-Guerin for injection came from BCG,this biological product can be identified by the detection of RD1.In our test,RD1 was shown to be present in M.bovis and the TB strain of H37Rv,but absent in the polysaccharide nucleic acid fraction of Bacillus Calmette-Guerin for injection and the strain that produces internal BCG vaccine for endermic injection with the same PCR product.So this method is fit for specific identification of polysaccharide nucleic acid of Bacillus Calmette-Guerin for injection.