国内外卡介菌的分子遗传学特性

目的比较我国生产用卡介菌上海菌株与国际常用卡介菌亚株(巴斯德1173P2株、法国F6株、丹麦1331株、丹麦2株、日本172株、Connaught)在分子遗传学上的差异。方法应用PCR和多重PCR方法,对国内外不同卡介菌的各缺失区(RD1、RD2、RD8、RD14、RD16、nRD18、RDDenmark/Glaxo)、插入序列IS6110拷贝数及基因座SenX3-Regx3串联重复序列拷贝数进行检测。结果我国生产用卡介菌上海菌株(BCG Chinese)缺失RD1和RD2,不缺失RD8、RD14、RD16、nRD18及RDDen-mark/Glaxo,含有1个IS6110插入序列,SenX3-Regx3基因座有3个串联重复序列,PCR产物为353bp;巴斯德1173P2株缺失RD1、RD2、RD14和nRD18,含有1个IS6110插入序列,SenX3-Regx3基因座有2个串联重复序列;法国F6株缺失RD1、RD2和nRD18,含有1个IS6110插入序列,SenX3-Regx3基因座有2个串联重复序列;丹麦1331株缺失RD1、RD2及RDDenmark/Glaxo,含有1个IS6110插入序列,SenX3-Regx3基因座有2个串联重复序列;丹麦2株与丹麦1331株相似,只是SenX3-Regx3基因座有2个串联重复序列和3个串联重复序列并存的现象;日本172株只缺失RD1,RD16只显示401bp的产物,含有2个IS6110插入序列;Connaught株缺失RD1、RD2、RD8及nRD18,含有1个IS6110插入序列,SenX3-Regx3基因座有1个串联重复序列。结论我国生产用卡介菌上海菌株与国外其他卡介菌亚株在分子遗传学特性上有一定的差异。

Molecular Genetic Characteristics of BCG Strains Used for Vaccine Production in China and Other Countries

Objective To compare the molecular genetic characteristics of BCG Chinese strain used for vaccine production in China and some BCG substrains used for vaccine production in other countries, such as Pasteur1173 P2, F6, Connaught, D1331, D2 and Jap172.Methods The regions of deletion(RD1, RD2, RD8, RD14, RD16, nRD18, RDDenmark / Glaxo)as well as copy numbers of inserted sequence IS6110 and locus SenX3-Regx3 tandem repeats of various BCG strains were determined by PCR and multiplex PCR.Results Shanghai strain showed a band of 353 bp on agarose gel electrophoretic profile and contained one copy of IS6110 and three repeats at SenX3-Regx3 locus, in which RD1 and RD2 were deleted while RD8, RD14, RD16, nRD18 and RDDen-mark / Glaxo were undeleted.Pasteur1173P2 strain contained one copy of IS6110 and two repeats at SenX3-Regx3 locus, in which RD1, RD2, RD14 and nRD18 were deleted.F6 strain contained one copy of IS6110 and two repeats at SenX3-Regx3 locus, in which RD1, RD2 and nRD18 were deleted.D1331 strain contain one copy of IS6110 and two repeats at SenX3-Regx3 locus, in which RD1, RD2 and RDDenmark / Glaxo were deleted.Like D1331 strain, D2 strain also contained one copy of IS6110, and its RD1, RD2 and RDDenmark / Glaxo were also deleted.However, the strain contained either two or three repeats at SenX3-Regx3 locus.Jap172 strain contained two copies of IS6110, in which RD1 was deleted and RD16 showed only a band of 401 bp.Connaught strain contained one copy of IS6110 and one repeat at SenX3-Regx3 locus, in which RD1, RD2, RD8 and nRD18 were deleted.Conclusion BCG Shanghai strain used for vaccine production in China showed a certain difference in molecular genetic characteristics with other BCG substrains used abroad.