A Simple and Efficient Multiplex PCR Assay for the Identification of Mycobacterium Genus and Mycobacterium tuberculosis Complex to the Species Level.
10.3349/ymj.2013.54.5.1220
- Author:
Yeun KIM
1
;
Yeonim CHOI
;
Bo Young JEON
;
Hyunwoo JIN
;
Sang Nae CHO
;
Hyeyoung LEE
Author Information
1. Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea. hyelee@yonsei.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't ; Validation Studies
- Keywords:
M. tuberculosis complex;
multiplex PCR;
rpoB;
RD1;
RD8
- MeSH:
Animals;
Cattle;
Classification/methods;
DNA Primers;
Genes, Bacterial;
Humans;
Multiplex Polymerase Chain Reaction/*methods;
Mycobacterium/classification/genetics/*isolation & purification;
Mycobacterium tuberculosis/classification/genetics/*isolation & purification;
Species Specificity
- From:Yonsei Medical Journal
2013;54(5):1220-1226
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: The Mycobacterium tuberculosis complex comprises M. tuberculosis, M. bovis, M. bovis bacillus Calmette-Guerin (BCG) and M. africanum, and causes tuberculosis in humans and animals. Identification of Mycobacterium spp. and M. tuberculosis complex to the species level is important for practical use in microbiological laboratories, in addition to optimal treatment and public health. MATERIALS AND METHODS: A novel multiplex PCR assay targeting a conserved rpoB sequence in Mycobacteria spp., as well as regions of difference (RD) 1 and RD8, was developed and evaluated using 37 reference strains and 178 clinical isolates. RESULTS: All mycobacterial strains produced a 518-bp product (rpoB), while other bacteria produced no product. Virulent M. tuberculosis complex strains, M. tuberculosis, M. bovis and M. africanum, produced a 254-bp product (RD1), while M. bovis BCG, M. microti and nontuberculous mycobacteria produced no RD1 region product. Additionally, M. tuberculosis and M. africanum produced a 150-bp product (RD8), while M. bovis and M. bovis BCG produced a 360-bp product (deleted form of RD8). M. microti and nontuberculous mycobacteria produced no RD8 region product. This assay identified all Mycobacterium spp. and all M. tuberculosis complex strains to the species level. CONCLUSION: The multiplex PCR assay of the present study could be implemented as a routine test in microbiology laboratories, and may contribute to more effective treatment and surveillance of tuberculosis stemming from the M. tuberculosis complex.
