Detection and genotyping of Mycobacterium species from clinical isolates and specimens by oligonucleotide array
Detection and genotyping of Mycobacterium species from clinical isolates and specimens by oligonucleotide array
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DOI：
10.1128/jcm.43.4.1782-1788.2005
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发表时间：
2005-04-01
影响因子：
9.4
通讯作者：
Kim, C
中科院分区：
文献类型：
--
作者：
Park, H;Jang, HJ;Kim, C
病原性分枝杆菌的鉴定对于分枝杆菌病的成功诊断是重要的。本研究的目的是建立一种利用分枝杆菌内转录间隔区（ITS）序列在种水平上检测和区分分枝杆菌的寡核苷酸芯片。利用一个属特异性探针和20个种特异性探针，包括两个M.利用禽-胞内复合物（MAC）特异性探针，我们开发了一种基于ITS的寡核苷酸芯片，用于快速可靠地检测和区分M。tuberculosis，MAC，M. fortupe，M. chelonae、龟类M. M.P. kansasii，M. gordonae、黑脉叶蝉M. scrofulaceum、M. szulgai，M. vaccae、牛痘M. xenopi，M. termium，M. flavescens，M. smegalgae，M. malmoense、M. simiae、M. marinum，M. ulcerans、M. gastri和M.麻风病人所有分枝杆菌均与属特异性探针（PAN-03）杂交，用于检测分枝杆菌属。除了分枝杆菌外，其他分枝杆菌与种特异性探针的杂交模式都是独特的. marinum和M.溃疡，其未被基于ITS的探针区分。在种特异性探针中，针对MAC存在多个亚种的情况，设计了两种特异性探针。使用46株参考菌株、149株临床分离株和155份临床标本证明了寡核苷酸阵列检测试剂盒的性能。整个过程（DNA提取、PCR、DNA杂交和扫描）在4.5 h内完成。我们的结果表明，寡核苷酸芯片是有用的鉴定和区分分枝杆菌从临床分离株和标本在普通的临床实验室。
Identification of pathogenic Mycobacterium species is important for a successful diagnosis of mycobacteriosis. The purpose of this study was to develop an oligonucleotide array which could detect and differentiate mycobacteria to the species level by using the internal transcribed spacer (ITS) sequence. Using a genus-specific probe and 20 species-specific probes including two M. avium-intracellulare complex (MAC)-specific probes, we have developed an ITS based oligonucleotide array for the rapid and reliable detection and discrimination of M. tuberculosis, MAC, M. fortuitum, M. chelonae, M. abscessus, M. kansasii, M. gordonae, M. scrofulaceum, M. szulgai, M. vaccae, M. xenopi, M. terrae, M. flavescens, M. smegmatis, M. malmoense, M. simiae, M. marinum, M. ulcerans, M. gastri, and M. leprae. All mycobacteria were hybridized with a genus-specific probe (PAN-03) for detection of the genus Mycobacterium. Mycobacterial species were expected to show a unique hybridization pattern with species -specific probes, except for M. marinum and M. ulcerans, which were not differentiated by ITS-based probe. Among the species-specific probes, two kinds of species-specific probes were designed for MAC in which there were many subspecies. The performance of the oligonucleotide array assay was demonstrated by using 46 reference strains, 149 clinical isolates, and 155 clinical specimens. The complete procedure (DNA extraction, PCR, DNA hybridization, and scanning) was carried out in 4.5 h. Our results indicated that the oligonucleotide array is useful for the identification and discrimination of mycobacteria from clinical isolates and specimens in an ordinary clinical laboratory.