Tuberc Respir Dis > Volume 58(2); 2005 > Article
Tuberculosis and Respiratory Diseases 2005;58(2):129-134.
DOI:    Published online February 1, 2005.
Detection of embB Gene Mutation of Mycobacterium tuberculosis by Reverse Hybridization Assay.
Young Kil Park, Hee Kyung Yu, Chan Hong Park, Sung Weon Ryu, Seung Heon Lee, Myung Sup Shim, Woo Jin Lew, Won Jung Koh, O Jung Kwon, Sang Nae Cho, Gill Han Bai
1Korean Institute of Tuberculosis, Seoul, Korea.
2Division of Pulmonary and Critical Care Medicine, Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
3Department of Microbiology, Yonsei University College of Medicine, Seoul, Korea.
Ethambutol (EMB) is one of important first-line drug in the treatment of tuberculosis. Molecular techniques to detect embB gene mutations have been considered as an method to define the EMB resistance. We investigated the mutation rate within embB gene among EMB resistant strains using reverse hybridization techniques. METHODS: We made 11 probes that had wild or mutated sequences containing codons 306, 406, or 497 within embB gene respectively. These probes were reverse-hybridized with PCR products amplified from embB gene which were isolated from 149 ethambutol resistant strains and 50 pan-susceptible strains. RESULTS: Out of 149 ethambutol resistant strains, one hundred (67.1%) had mutation at least one base at codon 306, 406, or 497 in embB gene. Mutation at codon 306, 406, 497 were demonstrated in 75 (50.3%), 16 (10.7%), and 13 strains (8.7%) respectively. There were four strains that showed multi-mutation at codon 306 and codon 406 simultaneously. A high proportion (8.1%) had single mutation at codon 406. There was no mutation observed in embB gene among 50 pan-susceptible strains. CONCLUSION: Reverse hybridization will be useful technique for detection of gene mutation correlated to ethambutol resistance.
Key Words: Mycobacterium tuberculosis, Genotypes, EmbB, Reverse hybridization, Ethambutol resistance

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