Title: Rapid Detection of rpoB Gene of Resistant Mycobacterium Tuberculosis in Sputum and Rifampin-resistant Mechanism
Abstract:Objective To rapidly detect rpoB gene of Mycobacterium tuberculosis in sputum and to explore rifampin(RIF)-resistant mechanism.MethodsrpoB fragment was amplified from M.tuberculosisstrains from sputum...Objective To rapidly detect rpoB gene of Mycobacterium tuberculosis in sputum and to explore rifampin(RIF)-resistant mechanism.MethodsrpoB fragment was amplified from M.tuberculosisstrains from sputum specimens of drug-resistant inpatients or clinic isolates by PCR with a pair of primers designed according to rifampin resistance-determining region.The fragments of rpoB were sequenced.The difference of rpoB gene fragments sequences of clinical isolates among multidrug-resistant,all drugs-resistant,single RIF-resistant,all drugs-susceptible isolates of M.tuberculosis,or standard strain were compared.ResultsWithin six hours,rpoB gene of M.tuberculosisstrains from sputum specimen of drug-resistant pulmonary tuberculosis was detected.By comparative analysis of gene sequence,spot mutation was found from multidrug-resistant and rifampin-resistant isolates in M.tuberculosisstrains.Codon 531,516 and 526 had higher mutational frequency in our RIF-resistant isolates.Only single nucleotide mutation was detected in most Rif-resistant isolates.But one multidrug-resistant isolate presented a rare double mutation at two codons of 531 and 479.No mutation in H37Rv standard strain or all drugs-susceptible isolates was detected.ConclusionrpoB gene and its mutation in M.tuberculosis strains could be rapidly and accurately identified by polymerase chain reaction and sequence analysis.RIF-resistance was related with spot mutation of rpoB gene in M.tuberculosisstrains.Read More
Publication Year: 2008
Publication Date: 2008-01-01
Language: en
Type: article
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