使用特异性rpoB寡核苷酸探针通过聚合酶链反应-连接反向杂交法鉴定结核分枝杆菌。

Identification of Mycobacterium tuberculosis by PCR-linked reverse hybridization using specific rpoB oligonucleotide probes.

作者信息

Hong Seong-Karp, Kim Bum-Joon, Yun Yeo-Jun, Lee Keun-Hwa, Kim Eui-Chong, Park Eun-Mi, Park Young-Gil, Bai Gil-Han, Kook Yoon-Hoh

机构信息

Department of Microbiology and Cancer Research Institute, Institute of Endemic Diseases, SNUMRC, Seoul National University College of Medicine, Chongno-Gu, Seoul 110-799, Republic of Korea.

出版信息

J Microbiol Methods. 2004 Oct;59(1):71-9. doi: 10.1016/j.mimet.2004.06.004.

DOI:10.1016/j.mimet.2004.06.004

PMID:15325754

Abstract

A reverse probe hybridization method using two different Mycobacterium tuberculosis-specific rpoB DNA probes in combination was evaluated for the identification of M. tuberculosis culture isolates. Among the 384 isolates tested, 354 strains were identified as M. tuberculosis, which included 37 rifampin-resistant strains, and 30 were nontuberculous mycobacteria (NTM). This result was in accord with partial rpoB sequence analysis and IS6110 polymerase chain reaction (PCR) results, but not with the results of biochemical testing, which produced two false negative results. Because of its high level of sensitivity and specificity, we suggest that M. tuberculosis-specific rpoB probes immobilized on micro-titer well plates or on other solid matrixes can be used efficiently for the rapid and convenient identification of M. tuberculosis.

摘要

评估了一种使用两种不同结核分枝杆菌特异性rpoB DNA探针组合的反向探针杂交方法，用于鉴定结核分枝杆菌培养分离株。在测试的384株分离株中，354株被鉴定为结核分枝杆菌，其中包括37株耐利福平菌株，30株为非结核分枝杆菌（NTM）。该结果与rpoB部分序列分析和IS6110聚合酶链反应（PCR）结果一致，但与生化检测结果不一致，生化检测产生了两个假阴性结果。由于其高灵敏度和特异性，我们建议固定在微量滴定板或其他固体基质上的结核分枝杆菌特异性rpoB探针可有效地用于快速、方便地鉴定结核分枝杆菌。