Tuberc Respir Dis.  2005 Feb;58(2):129-134.

Detection of embB Gene Mutation of Mycobacterium tuberculosis by Reverse Hybridization Assay

• Park YK
• Yu HK
• Park CH
• Ryu SW
• Lee SH
• Shim MS
• Lew WJ
• Koh WJ
• Kwon OJ
• Cho SN
• Bai GH

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.

Keyword

Mycobacterium tuberculosis; Genotypes; EmbB; Reverse hybridization; Ethambutol resistance

MeSH Terms

Codon
Ethambutol
Genotype
Mutation Rate
Mycobacterium tuberculosis*
Mycobacterium*
Polymerase Chain Reaction
Tuberculosis
Codon
Ethambutol