[Evaluation of pyrosequencing for the detection of rpoB gene mutation in Mycobacterium tuberculosis].
OBJECTIVE
To detect the mutations of rpoB gene in Mycobacterium tuberculosis by pyrosequencing and to evaluate the values on detection of rifampin resistance in clinical isolates.


METHODS
Using the new technology of pyrosequencing, the mutations in the rifampin resistance determining region (RRDR) of rpoB gene were analyzed. The results were compared with those obtained from methods of the absolute concentration and the minimum inhibitory concentration (MIC).


RESULTS
Among the 150 Mycobacterium tuberculosis clinical isolates, 84 were susceptible and 66 resistant to RIF. 54 of the 66 resistant isolates were multidrug-resistant (MDR) strains. Ser531Leu and His526Asp or Tyr, including twelve different genotypes and six codons, were the most common mutations. In the drug susceptibility testing, the accordance rates of the pyrosequencing and the absolute concentration method as well as MIC were 92.7% and 97.8% respectively.


CONCLUSIONS
Not only is the pyrosequencing technology a fast, sensitive and high throughput method in detecting rifampin resistance in Mycobacterium tuberculosis, but also a useful tool in the research of rifampin resistance mechanism.
Zheng
Rui-juan
RJ
Shanghai Key Laboratory of Tuberculosis, Shanghai Pulmonary Hospital, Affiliated Tongji University, Shanghai 200433, China.
Wang
Jie
J
Qin
Lian-hua
LH
Lu
Jun-mei
JM
Hu
Zhong-yi
ZY
D001426:Bacterial Proteins / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D006801:Humans; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D010756:Phosphoric Acids; D016133:Polymerase Chain Reaction; D012293:Rifampin / Q000494:pharmacology*
D001426:Bacterial Proteins; D010756:Phosphoric Acids; C505805:rpoB protein, Mycobacterium tuberculosis; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin