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首页> 外文期刊>Journal of Clinical Microbiology >Detection and Genotyping of Mycobacterium Species from Clinical Isolates and Specimens by Oligonucleotide Array
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Detection and Genotyping of Mycobacterium Species from Clinical Isolates and Specimens by Oligonucleotide Array

机译：寡核苷酸阵列从临床分离物和标本中检测和分枝杆菌种的基因分型

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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.
机译：致病性分枝杆菌物种的鉴定对于成功诊断分枝杆菌病很重要。这项研究的目的是开发一种寡核苷酸阵列，该阵列可通过使用内部转录间隔区（ITS）序列将分枝杆菌检出并区分到物种水平。使用属特异性探针和20种物种特异性探针，其中包括两个 M。鸟细胞内复合物（MAC）特异性探针，我们开发了一种基于ITS的寡核苷酸阵列，可快速可靠地检测和区分 M。结核病，MAC， M。 ， M。 chelonae ， M。脓肿， M。堪萨斯州， M。 gordonae ， M。 s ， M。 szulgai ， M。 vaccae ， M。 xenopi ， M。 terrae ， M。 flavescens ， M。包皮垢， M。马尔摩斯， M。 simiae ， M。 marinum ， M。溃疡， M。 gastri 和 M。麻风病。将所有分枝杆菌与一种属特异性探针（PAN-03）杂交，以检测分枝杆菌属。除 M之外，预期分枝杆菌属物种会显示出与物种特异性探针的独特杂交模式。 marinum 和 M。溃疡，无法通过基于ITS的探针进行区分。在种特异性探针中，为MAC设计了两种种特异性探针，其中存在许多亚种。通过使用46株参考菌株，149株临床分离株和155株临床标本证明了寡核苷酸阵列测定的性能。整个过程（DNA提取，PCR，DNA杂交和扫描）在4.5小时内完成。我们的结果表明，在常规临床实验室中，寡核苷酸阵列可用于从临床分离物和标本中鉴定和鉴别分枝杆菌。

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