Detection of streptomycin-resistance associated rpsL and rrs gene mutations in Mycobacterium tuberculosis by PCR-single-strand-conformational polymorphism
10.3760/cma.j.issn.1674-2397.2011.05.007
- VernacularTitle:应用PCR-SSCP技术检测结核分枝杆菌耐链霉素相关基因rpsL和rrs突变
- Author:
Zhaodong LI
;
Hui WEI
;
Dapeng FAN
;
Peng DU
;
Aihua SUN
- Publication Type:Journal Article
- Keywords:
Mycobacterium tuberculosis;
Polymerase chain reaction-single-strand conformational polymorphism;
rpsL gene;
rrs gene;
Streptomycin;
Drug resistance;
Mutation
- From:
Chinese Journal of Clinical Infectious Diseases
2011;04(5):275-277
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo establish a novel rapid detection method based on PCR-single-strand-conformational polymorphism (PCR-SSCP) to determine mutation of streptomycin-resistance associated rpsL and rrs genes in isolates of Mycobacterium tuberculosis (MTB).MethodsStreptomycin-resistance of 112 MTB isolates was detected using the routine drug susceptibility test,and a special PCR-SSCP assay was established.The mutations of rpsL and rrs genes in streptomycin-resistant MTB isolates were detected by PCR-SSCP and PCR direct sequencing (PCR-DS) ; the results from two techniques were compared.Results All isolates had both rpsL and rrs genes.Fifty-two isolates (46.4％) were streptomycin susceptible,in which only 1 isolate showed abnormal PCR-SSCP fragments from rrs gene,and the specificity of PCR-SSCP was 98.1％ (51/52).Sixty isolates (53.6％) were streptomycin-resistant,in which 46 (76.6％) and 11 ( 18.3％ ) isolates presented the abnormal PCR-SSCP fragments of rpsL and rrs gene,respectively.One streptomycin-resistant isolate showed abnormal PCR-SSCP fragments from both rpsL and rrs genes.The sensitivity of PCR-SSCP was 93.3％ (56/60).ConclusionThe PCR-SSCP that established in this study is a specific and sensitive method for rapid detection of the streptomycin-resistance associated mutations in rpsL and rrs genes of MTB.
