利用PCR结合膜芯片技术检测结核分支杆菌的实验研究

THE EXPERIMENTAL STUDY OF DETECTING MYCOBACTERIUM TUBERCULOSIS BY PCR AND GENE MEMBRANE CHIP

目的:探讨聚合酶链反应(PCR)结合膜芯片技术用于检测结核分支杆菌的方法。方法:标记生物素的特异性引物,扩增17株菌标本IS6110插入序列的245bp目的基因,结合膜芯片杂交检测结核分支杆菌基因。结果:15株结核分支杆菌标本中有12株扩增出245bp目的基因,2株大肠杆菌及阴性对照标本PCR扩增结果为阴性。经膜芯片杂交,12株结核分支杆菌标本中有11株显色结果为阳性,其余显色结果为阴性。PCR结合膜芯片检测结核分支杆菌的灵敏度为73.3%,特异度为100%。结论:PCR结合膜芯片技术是一种具有发展潜力的检测结核分支杆菌的方法。

Objective. To establish PCR and microarray primely for detecting mycobacterium tuberculosis （MTB）. Methods. Primers marked by biotin amplified IS6110 245 bp in 17 bacteria strains,and microarray detected MTB. Result：The 245 bp of 12 in 15 MTB strains were amplified, and the results of 2 E. coli strains and negative control sample were negative. After hybridization by microarray, the results of 11 in coloured 12 MTB strains were positive, while the other results were negative. The sensitivity and the specificity was separately 73.3% and 100% by PCR and microarray detecting MTB. Conclusion. PCR and microarray will be a hopefully potential method for detecting MTB.