Characterization of rpoB mutations in rifampin-resistant clinical Mycobacterium tuberculosis isolates from Kuwait and Dubai.
Mutations conferring resistance to rifampin in rifampin-resistant clinical Mycobacterium tuberculosis isolates occur mostly in the 81 bp rifampin-resistance-determining region (RRDR) of the rpoB gene. In this study, 29 rifampin-resistant and 12 -susceptible clinical M. tuberculosis isolates were tested for characterization of mutations in the rpoB gene by line probe (INNO-LiPA Rif. TB) assay and the results were confirmed and extended by DNA sequencing of the PCR amplified target DNA. The line probe assay identified all 12 susceptible strains as rifampin-sensitive and the DNA sequence of RRDR in the amplified rpoB gene from two isolates matched perfectly with the wild-type sequence. The line probe assay identified 28 resistant isolates as rifampin-resistant with specific detection of mutation in 22 isolates including one isolate that exhibited hetro-resistance containing both the wild-type pattern as well as a specific mutation within RRDR while one of the rifampin-resistant strain was identified as rifampin-susceptible. DNA sequencing confirmed these results and, in addition, led to the specific detection of mutations in 5 rifampin-resistant isolates in which specific base changes within RRDR could not be determined by the line probe assay. These analyses identified 8 different mutations within RRDR of the rpoB gene including one novel mutation (S522W) that has not been reported so far. The genotyping performed on the isolates carrying similar mutations showed that majority of these isolates were unique as they exhibited varying DNA banding patterns. Correlating the ethnic origin of the infected TB patients with the occurrence of specific mutations at three main codon positions (516, 526 and 531) in the rpoB gene showed that most patients (11 of 15) from South Asian region contained mutations at codon 526 while majority of isolates from patients (6 of 11) of Middle Eastern origin contained mutations at codon 531.
Ahmad
Suhail
S
Department of Microbiology, Faculty of Medicine, Kuwait University, P.O. Box 24923, 13110, Safat, Kuwait. suhail_ah@hs.kuniv.edu.kw
Mokaddas
Eiman
E
Fares
Esther
E
D000904:Antibiotics, Antitubercular / Q000494:pharmacology*; D016172:DNA Fingerprinting / Q000379:methods; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D004352:Drug Resistance, Microbial; D005260:Female; D005838:Genotype; D006801:Humans; D007730:Kuwait / Q000453:epidemiology / Q000208:ethnology; D008297:Male; D008826:Microbial Sensitivity Tests; D008877:Middle East / Q000453:epidemiology / Q000208:ethnology; D008969:Molecular Sequence Data; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000145:classification / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods; D012293:Rifampin / Q000494:pharmacology*; D017422:Sequence Analysis, DNA; D014397:Tuberculosis, Pulmonary / Q000453:epidemiology* / Q000208:ethnology / Q000382:microbiology
D000904:Antibiotics, Antitubercular; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit; D012293:Rifampin