title: [Detecting rpoB gene mutation in rifampin-resistant Mycobacterium tuberculosis by using the reverse dot-blot hybridization method].
abstract: OBJECTIVE
  To establish a simple, rapid and sensitive hybridization method for detecting drug-resistance relevant gene mutation in Mycobacterium tuberculosis.
  
  
  METHODS
  Fourteen single-strand specific probes designed to detect mutated and/or wild rpoB gene in Mycobacterium tuberculosis were spotted and fixed on nylon membranes, and PCR products labeled with biotin were obtained by using down-stream primer labeled with biotin, then hybridized and analyzed with streptavidin-HRP and TMB.
  
  
  RESULTS
  Twenty-three rifampin-resistant Mycobacterium tuberculosis isolates and 11 rifampin-sensitive isolates were analyzed. The gene mutations were consistent with the DNA sequencing and the in vitro susceptibility test in 30/34 and 28/34 of the isolates, respectively. Mutations of Ser-531 were present in 11 of the 23 rifampin-resistant isolates, followed by His-526, Leu-533, and no mutation was found in 13 isolates, including 2 rifampin-resistant isolates. Mutations in loci 516 and 513 were not detected.
  
  
  CONCLUSIONS
  Reverse dot-blot hybridization may be of potential use for the rapid diagnosis of rifampin-resistant tuberculosis.
authors.affiliation: Department of Microbiology, Second Military Medical University, Shanghai 200433, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Wang
authors.forename: Wei
authors.initials: W
authors.lastname: Pan
authors.forename: Weirong
authors.initials: W
authors.lastname: Jin
authors.forename: Xinhua
authors.initials: X
authors.lastname: Weng
authors.forename: Ran
authors.initials: R
authors.lastname: Yan
authors.forename: Bei
authors.initials: B
authors.lastname: Su
authors.forename: Shu
authors.initials: S
authors.lastname: Chen
authors.forename: Wenhong
authors.initials: W
authors.lastname: Zhang
authors.forename: Hongzhou
authors.initials: H
authors.lastname: Lu
authors.forename: Zhongtian
authors.initials: Z
authors.lastname: Qi
mesh_terms: D000904:Antibiotics, Antitubercular / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000737:chemistry; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D012293:Rifampin / Q000494:pharmacology*
chemical_list: D000904:Antibiotics, Antitubercular; D004269:DNA, Bacterial; D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit; D012293:Rifampin