Real-time fluorescence PCR-molecular beacon assay for detecting mutations in rpoB genes of rifampin-resistant Mycobacterium tuberculosis
- VernacularTitle:实时荧光PCR分子信标检测耐利福平结核分枝杆菌印rpoB基因
- Author:
Guizhi SUN
;
Tiejie GAO
;
Haohao ZHONG
;
Lijun KANG
;
Zhiguo ZHANG
;
Wanjie HENG
;
Bingquan WU
;
Wei LIU
- Publication Type:Journal Article
- Keywords:
Polymerase chain reaction;
Mycobactsrium tuberculosis;
Bacterial proteins;
Mutation;
Rifampin;
Drug resistance,bactenial
- From:
Chinese Journal of Laboratory Medicine
2008;31(4):429-432
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a rapid method to detect mutations in rpoB genes of rifampin-resistant Mycobacterium tubereulosis in dinical specimens using Real-time fluorescence PCR molecular beacon assay.Methods 174 strains of Mvcobacterium tuberculosis clinical isolates were analyzed using real-time fluorescence PCR molecular beacon assay foilowed with DNA sequencing while 12 strains of NTM and 4 strains of bacteria other than Mycobacterium tuberculosis were used as the contrast.Results Eighty-two 89.1 of 92 rifampin (RIF)-resistant strains and 3 of 82 RIF-sensitive strains were found to harbor mutation in the rpoB gene using real-time fluorescence PCR-molecular beacon assay.The specificity, sensitivity,and accuracy of this assay were 96.3%,89.1%,and 92.5%,respectively-Eithty-three of 92 RIF-resistant strains and 1 of 82 RIF-sensitive strains were found to harbor mutation in the rpoB gene using the direct DNA sequencing.The specificity,sensitivity,and accuracy of the direct DNA sequencing were 98.8,90.2%,and 94.2%,respectively.As compared with real-time PCR molecular beacon assay,171 of 174(98.3%)strains of myeobactefium tuberculosis clinical isolates had the salne results.Conclusion Real-time fluorescence PCR-molecular beacon assay can be used as a rapid screen method to detect RIF-resistant isolates.
