역교잡 방법을 이용한 결핵균 embB 유전자 돌연변이 검출
Detection of embB Gene Mutation of Mycobacterium tuberculosis by Reverse Hybridization Assay
권오정 (성균관대학교)
류성원 (대한결핵협회 결핵연구원)
류우진 (대한결핵협회 결핵연구원)
고원중 (성균관대학교)
조상래 (연세대학교)
배길한 (대한결핵협회 결핵연구원)
유희경 (결핵연구원)
박찬홍 (결핵연구원)
이승헌 (결핵연구원)
심명섭 (결핵연구원)
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Abstract
Background : 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. (Tuberc Respir Dis 2005; 58:129-134)
- keywords
- Mycobacterium tuberculosis, Genotypes, EmbB, Reverse hybridization, Ethambutol resistance
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