利用rpoB基因芯片技术进行快速分枝杆菌菌种鉴定

Rapid Species Identification of Mycobacterium tuberculosis by rpoB Gene Micro Array

利用rpoB基因芯片技术快速进行分枝杆菌菌种鉴定。以分枝杆菌rpoB基因编码序列为靶基因,用基因芯片技术检测21种分枝杆菌标准株;8种其它细菌标准株;126株临床分离株。分枝杆菌与其它细菌标准株经PCR扩增后,分枝杆菌标准株均扩增出360 bp DNA片段,在其它细菌中,除甲型溶血性链球菌和假白喉棒状杆菌出现同样片段外,其它细菌均未见扩增。21种寡核苷酸探针除海分枝杆菌与偶然分枝杆菌的探针有交叉杂交外,其余均为特异性杂交。对126株临床分离株进行鉴定,89株为结核分枝杆菌,占70.6%(89/126),非结核分枝杆菌(NTM)占9.2%(9/98)。应用rpoB基因芯片技术鉴定分枝杆菌菌种,是一种快速、准确的方法,具有较高的临床应用价值。

Rapid species identification of Mycobacterium tuberculosis by rpoB gene micro array. Based on the gene micro array of rpoB, the standard strains of 21 mycobacteria and 8 non-mycobacteria ,126 clinical isolated of mycobacteria were detected by PCR-reverse dot blot hybridization assay. 360bp DNA fragment was amplified from all mycobacteria tested and was not found in all nonmycobacteria except Hemolytic streptococcus and Corynebacterium pseudodiphtheriticum. The result of specimens were detected by the probe which is composed of 21 oligonucleotide was that probe-Mycobacterium fortuitum cross hybridizated with Mycobacterium platypoecilus while the other probes were specific. The 89 strains of the all 126 strains isolated from clinical specimens were identified to be mycobacterium tuberculosis, the percentage was 70.6%（89/126）, while the other 9 strains were identified to be unmycobacterium tuberculosis and the the percentage was 9.2%（9/98）. Identification of Mycobacteria by rpoB gene micro array is a rapid and effective method which is of considerable value in clinical territory.