Application of Gene Chip for Detecting Mutations of Rifampin-resistant Genes in Mycobacterium Tuberculosis Strains / 中国医师杂志

ABSTRACT

Objective To explore the feasibility of detecting the mutations of rpoB gene in rifampin-resistant Mycobacterium tuberculosis strains by gene chip. Methods The DNA chip was designed according to the sequence of Mycobacterium tuberculosis rpoB gene. The DNA fragments which contained hot mutation sites of rpoB gene were labelled with cy3 fluorescence, amplified by PCR technique and hybridized with gene chip. The DNA sequencing was used as the control. Results Eighteen of the Mycobacterium tuberculosis strains were detected for the mutations of rpoB gene by gene chip. Among them, one sensitive strain was identical with the type strain, and all of 17 strains with rifampin-resistant, rpoB mutations were detected. Of the 17 strains with rpoB mutations, 14 were detected as single point mutation, one was detected as double sites point mutations with the sites of 511 and 516, and two were detected as single point mutation with the sites of 518 and 526, respectively. Because the probe of 517 rifampin-resistant did not be dotted on the gene chip, only sites of 518 and 526 mutations were detected, which were consistent with the results of DNA sequencing. Conclusion Using the gene chip could detect mass rpoB gene mutation of rifampin-resistant with higher specialty and sensitivity, which could be used for clinical detection of rifampin-resistant strains and clinical medicine.