Cheunoy Wattana, Prammananan Therdsak, Chaiprasert Angkana, Foongladda Suporn
Department of Microbiology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Diagn Microbiol Infect Dis. 2005 Mar;51(3):165-71. doi: 10.1016/j.diagmicrobio.2004.09.006.
The increasing incidence of tuberculosis and other mycobacterial infections due to AIDS epidemic resulted in the need of rapid and accurate identification of isolated mycobacteria. The correct identification result leads to the selection of an appropriate therapeutic regimen. Polymerase chain reaction and restriction enzyme analysis (PCR-REA) has been developed since 1992 and used as the rapid method for identifying mycobacteria. Several genes or sequences have been used as an amplified target for PCR-REA. The present study aims to evaluate the potential use of PCR-REA of gene-encoding heat shock protein 65 kDa (hsp65) and beta-subunit RNA polymerase (rpoB) for the identification of mycobacteria compared with conventional biochemical identification. Two hundreds clinical isolates, consisting of 50 isolates of Mycobacterium tuberculosis and 150 isolates of nontuberculous mycobacteria (NTM), were submitted for identification using PCR-REA and biochemical method. The results demonstrated that PCR-REA identified 188 isolates of both M. tuberculosis and NTM concordantly with biochemical identification. Discordant identification results obtained from 12 isolates, comprised of 8 M. scrofulaceum, 1 M. avium complex, 1 M. malmoense, 1 M. terrae complex, and 1 M. chelonae/abscessus. Overall, the concordant percentage of results obtained from PCR-REA compared with biochemical method was 100%, 98.8%, and 83.3% for M. tuberculosis complex, rapidly growing, and slowly growing mycobacteria, respectively, and the results of hsp65 PCR-REA was in agreement with those obtained from rpoB PCR-REA. From this study, PCR-REA appears to be a simple, rapid, and reliable method for identifying mycobacteria in a routine microbiology laboratory.
由于艾滋病流行导致结核病和其他分枝杆菌感染的发病率不断上升，因此需要快速准确地鉴定分离出的分枝杆菌。正确的鉴定结果有助于选择合适的治疗方案。聚合酶链反应和限制性酶切分析（PCR-REA）自1992年以来得到发展，并被用作鉴定分枝杆菌的快速方法。几个基因或序列已被用作PCR-REA的扩增靶点。本研究旨在评估编码65 kDa热休克蛋白（hsp65）和β亚基RNA聚合酶（rpoB）的基因的PCR-REA与传统生化鉴定相比在分枝杆菌鉴定中的潜在用途。提交了200株临床分离株进行鉴定，其中包括50株结核分枝杆菌和150株非结核分枝杆菌（NTM），采用PCR-REA和生化方法进行鉴定。结果表明，PCR-REA鉴定出的188株结核分枝杆菌和NTM与生化鉴定结果一致。12株分离株的鉴定结果不一致，包括8株瘰疬分枝杆菌、1株鸟分枝杆菌复合群、1株马尔默分枝杆菌、1株地分枝杆菌复合群和1株龟分枝杆菌/脓肿分枝杆菌。总体而言，与生化方法相比，PCR-REA对结核分枝杆菌复合群、快速生长和缓慢生长分枝杆菌的鉴定结果一致率分别为100%、98.8%和83.3%，hsp65 PCR-REA的结果与rpoB PCR-REA的结果一致。从本研究来看，PCR-REA似乎是一种在常规微生物实验室中鉴定分枝杆菌的简单、快速且可靠的方法。
