Comparative evaluation of polymerase chain reaction and restriction enzyme analysis: Two amplified targets, hsp65 and rpoB, for identification of cultured mycobacteria
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 reacti...
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th-mahidol.171312018-06-21T15:32:08Z Comparative evaluation of polymerase chain reaction and restriction enzyme analysis: Two amplified targets, hsp65 and rpoB, for identification of cultured mycobacteria Wattana Cheunoy Therdsak Prammananan Angkana Chaiprasert Suporn Foongladda Mahidol University Thailand National Center for Genetic Engineering and Biotechnology Medicine 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 β-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. © 2005 Elsevier Inc. All rights reserved. 2018-06-21T08:32:08Z 2018-06-21T08:32:08Z 2005-01-01 Article Diagnostic Microbiology and Infectious Disease. Vol.51, No.3 (2005), 165-171 10.1016/j.diagmicrobio.2004.09.006 07328893 2-s2.0-17644411689 https://repository.li.mahidol.ac.th/handle/123456789/17131 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=17644411689&origin=inward |
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Medicine Wattana Cheunoy Therdsak Prammananan Angkana Chaiprasert Suporn Foongladda Comparative evaluation of polymerase chain reaction and restriction enzyme analysis: Two amplified targets, hsp65 and rpoB, for identification of cultured mycobacteria |
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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 β-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. © 2005 Elsevier Inc. All rights reserved. |
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Mahidol University |
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Mahidol University Wattana Cheunoy Therdsak Prammananan Angkana Chaiprasert Suporn Foongladda |
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Article |
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Wattana Cheunoy Therdsak Prammananan Angkana Chaiprasert Suporn Foongladda |
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Wattana Cheunoy |
title |
Comparative evaluation of polymerase chain reaction and restriction enzyme analysis: Two amplified targets, hsp65 and rpoB, for identification of cultured mycobacteria |
title_short |
Comparative evaluation of polymerase chain reaction and restriction enzyme analysis: Two amplified targets, hsp65 and rpoB, for identification of cultured mycobacteria |
title_full |
Comparative evaluation of polymerase chain reaction and restriction enzyme analysis: Two amplified targets, hsp65 and rpoB, for identification of cultured mycobacteria |
title_fullStr |
Comparative evaluation of polymerase chain reaction and restriction enzyme analysis: Two amplified targets, hsp65 and rpoB, for identification of cultured mycobacteria |
title_full_unstemmed |
Comparative evaluation of polymerase chain reaction and restriction enzyme analysis: Two amplified targets, hsp65 and rpoB, for identification of cultured mycobacteria |
title_sort |
comparative evaluation of polymerase chain reaction and restriction enzyme analysis: two amplified targets, hsp65 and rpob, for identification of cultured mycobacteria |
publishDate |
2018 |
url |
https://repository.li.mahidol.ac.th/handle/123456789/17131 |
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1763487308171968512 |