Title: Molecular analysis of rpoB gene mutations of rifampin-resistant Mycobacterium tuberculosisclinical isolates in Xi'an City
Abstract:Objective To genotype the rifampin(RFP)-resistance-determining region(RRDR) of rpoB gene of clinical Mycobacterium tuberculosis isolates in Xi'an by PCR single-strand conformation polymorphism(PCR-SSC...Objective To genotype the rifampin(RFP)-resistance-determining region(RRDR) of rpoB gene of clinical Mycobacterium tuberculosis isolates in Xi'an by PCR single-strand conformation polymorphism(PCR-SSCP) combined with DNA sequencing.Methods The rpoB gene of 32 RFP-resistant and 10 RFP-susceptible Mycobacterium tuberculosis strains was analyzed by PCR-SSCP within the 157 bp(Ala500 to Val550) region covers RRDR,and the PCR products of rpoB gene of 8 strains,which have representative patterns of PCR-SSCP combined with drug sensitive testing,were sequenced subsequently.Results The sensitivity and specificity of PCR-SSCP in detected rpoB gene were 56.3%(18/32) and 80%(8/10),respectively.Mutations in codon 531 or 526 were observed in 5 of 6 RFP-resistant isolates by DNA sequencing,and double-base mutations in codon 526 were found in 2 of 6 RFP-resistant isolates.One mutation in codon 513 with false-negative PCR-SSCP result was also unfolded.Two false-positive PCR-SSCP results were revealed among the RFP-susceptible strains with mutations covered Leu511,Asp516 and Asn518,in which new asparagine 518 mutation type was identified.Conclusion The mutations in codon 531 and codon 526 of rpoB gene are still the hot spots.Both point mutant and double mutants in one codon were discovered.Moreover,multiple mutations in codons of RFP-susceptible isolates deserve concern.Read More
Publication Year: 2012
Publication Date: 2012-01-01
Language: en
Type: article
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