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首页> 外文期刊>Journal of Clinical Microbiology >Detection and identification of mycobacteria by DNA amplification and oligonucleotide-specific capture plate hybridization.
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Detection and identification of mycobacteria by DNA amplification and oligonucleotide-specific capture plate hybridization.

机译：通过DNA扩增和寡核苷酸特异性捕获板杂交检测和鉴定分枝杆菌。

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We have developed an easy and rapid detection and identification system for the diagnosis of mycobacterial diseases. The system is based on selective amplification by PCR of mycobacteria with primers based on the genes coding for 16S rRNA. During PCR, a label (digoxigenin-11-dUTP) is incorporated with biotinylated species-specific oligonucleotides (oligonucleotide-specific capture plate hybridization [OSCPH]. One oligonucleotide specific for the genus Mycobacterium and seven species-specific (Mycobacterium tuberculosis, M. avium, M. intracellulare, M. scrofulaceum, M. xenopi, M. genavense, and M. chelonae) oligonucleotides were designed as capturing probes. After specific hybridization, an enzyme immunoassay reveals the specifically bound complexes and thus permits identification of the mycobacterium. A total of 70 mycobacterial strains were tested. For 69 strains, results concordant with conventional identification were obtained. One M. chelonae strain was negative with the M. chelonae probe and was later reidentified as M. fortuitum. Moreover, for 15 clinical samples suspected of harboring nontuberculous mycobacteria, OSCPH was able to confirm all culture results and could identify one M. genavense infection for which standard culture results were negative. PCR-OSCPH is easily applicable and much faster than culture. It could become a valuable alternative approach for the diagnosis of mycobacterial infections.
机译：我们已经开发了一种简便，快速的检测和鉴定系统，用于诊断分枝杆菌疾病。该系统基于通过使用基于编码16S rRNA的基因的引物对分枝杆菌进行的PCR选择性扩增。在PCR期间，将标记（digoxigenin-11-dUTP）与生物素化的物种特异性寡核苷酸（寡核苷酸特异性捕获板杂交[OSCPH]。一种分枝杆菌属特异性的寡核苷酸和七种物种特异性（结核分枝杆菌，鸟分枝杆菌） （例如，M.intracellulare，M.scrofulaceum，M.xenopi，M.genavense和M. chelonae）寡核苷酸被设计成捕获探针，经过特异性杂交后，酶联免疫法揭示了特异性结合的复合物，因此可以鉴定分枝杆菌。总共测试了70株分枝杆菌菌株，其中69株获得了与常规鉴定结果相符的结果，其中1株M. chelonae菌株经M. chelonae探针鉴定为阴性，后来被重新鉴定为M. fortuitum，此外，还对15株临床疑似带有非结核分枝杆菌的OSCPH能够确认所有培养结果，并可以鉴定出一种标准培养结果为阴性。 PCR-OSCPH易于使用，并且比培养要快得多。它可能成为诊断分枝杆菌感染的有价值的替代方法。

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