PCR测序和基因芯片快速鉴定分枝杆菌菌种的应用研究

Comparison of PCR sequencing and DNA microarray for rapid identification of mycobacterium species

目的评价2种分枝杆菌菌种快速鉴定方法的临床应用价值。方法分别用16S～23SrDNA转录间隔序列(ITS)PCR-直接测序和基于16SrRNA基因的基因芯片检测21株分枝杆菌标准菌株和50株临床分离株。结果 21株分枝杆菌标准株中,ITS PCR测序法将18株准确鉴定到种,结核分枝杆菌、非洲分枝杆菌菌株只能鉴定到结核分枝杆菌复合群,海分枝杆菌无法鉴别是海分枝杆菌还是溃疡分枝杆菌;芯片法正确鉴定16株,1株胃分枝杆菌鉴定为堪萨斯分枝杆菌,3株不在芯片检测范围内。50株临床菌株中,47株2种方法结果一致,1株不在芯片检测范围内。结论基因芯片鉴定分枝杆菌快速、特异,准确性高,临床常规开展应用需要进一步研究。

Objective To evaluate the value on rDNA sequencing of 16S-23S internal transcription space （ITS） and DNA microarray based on 16S rDNA for the rapid identification of mycobacterial species. Methods 21 mycobacterium reference strains and 50 clinical isolates were simultaneously identified by DNA sequencing and DNA microarray. Results Of 21 mycobacterium reference strains, 18 were identified correctly at species level by DNA sequencing. M. africanum and M. tuberculosis strains were identified as M. tuberculosis complex, and M. marinum strain was identified as M. marinum/M, ulcerans. 16 strains were identified correctly by DNA microarray, but 1 M. gastri strain was identified as M. kansasii, 3 strains were absent of the specific probes in the microarray. Of 50 clinical isolates, 47 were consistent by DNA sequencing and DNA microarray, and one M. lentiflavum isolates was not identified by DNA microarray due to the absence of the specific probe. Conclusion DNA microarray was a specific, accurate method for the rapid identification of most clinically relevant mycobacteria.