Rapid detection of MDR-Mycobacterium tuberculosis using modified PCR-SSCP from clinical Specimens
10.12980/APJTB.4.2014C1186
- Author:
Babak Farzam
;
Fazlollah Seyed Mousavi
;
Nematollah Jafari Jonaidi
- Publication Type:Journal Article
- Keywords:
Drug resistance;
Mycobacterium tuberculosis;
PCR-SSCP;
Phenotypic method;
rpoB;
katG
- From:Asian Pacific Journal of Tropical Biomedicine
2014;(z1):165-170
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To design a rapid test to detect the rifampin (RIF) and isoniazid (INH) resistant mutant based on polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique that analyzes the katG, rpoB genes.Methods:tuberculosis. To determine the susceptibility of isolates to anti TB drugs, the proportional method was used. Mutations presented within the amplified products of the katG, rpoB genes were evaluated by SSCP.Results:Using proportional method, 12 (11.6%) and 9 (8.7%) isolates were resistant respectively Biochemical test as well as IS6110 targeting PCR revealed 103 clinical samples were to INH and RIF and 9 (8.7%) isolates showed resistance to both drug (multi-drug resistant tuberculosis). Three (2.9%) multi-drug resistant tuberculosis and two INH resistant isolates were detected by the PCR-SSCP and sequencing. The sensitivity and specificity of PCR-SSCP for multi-drug resistant isolates were 33% and 100%, respectively.Conclusions:Complete agreement between SSCP and sequencing can indicate that resistance-associated mutations have occurred in other genes except our considered genes.