Establishment and drug susceptibility test of isoniazid resistant Mycobacterium smegmatis.
- Author:
Ping-ping JIA
1
;
Li-li ZHAO
;
Xiao-yu LI
;
Quan ZHANG
;
Zhen-long LIU
;
Xin WANG
;
Li-yan YU
;
Li-xun ZHAO
;
Shan CEN
Author Information
1. Laboratory of Immunobiology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing 100050, China.
- Publication Type:Journal Article
- MeSH:
Anti-Bacterial Agents;
pharmacology;
Antibiotics, Antitubercular;
pharmacology;
Antitubercular Agents;
pharmacology;
Bacterial Proteins;
genetics;
metabolism;
Drug Resistance, Bacterial;
Electroporation;
Ethambutol;
pharmacology;
Isoniazid;
pharmacology;
Microbial Sensitivity Tests;
Mycobacterium smegmatis;
drug effects;
genetics;
metabolism;
Oxidoreductases;
genetics;
metabolism;
Plasmids;
Rifampin;
pharmacology;
Streptomycin;
pharmacology
- From:
Acta Pharmaceutica Sinica
2011;46(4):377-382
- CountryChina
- Language:Chinese
-
Abstract:
With the emergence of drug resistant tuberculosis, it is very urgent to find novel anti-tuberculosis drugs, especially novel anti-drug-resistant tuberculosis drugs. Because of the slow growth and the need to work in a biosafty environment of Mycobacterium tuberculosis, the development of evaluation of drug effect is severely impeded. In order to solve these issues, non-pathogenic fast-growing Mycobacterium smegmatis is introduced as test organism. The inhA is one of a target of isoniazid (INH) overexpression or mutation of this gene in Mycobacterium tuberculosis conferring resistant to INH. A recombinant plasmid bearing inhA was constructed and electroporated into Mycobacterium smegmatis, using shuttle expression vector pMV261. Transformants were induced to express a protein of inhA, identified by SDS-PAGE. Results show that Mycobacterium smegmatis containing inhA plasmids exhibited 100-fold or greater increased resistance to INH, but it conferred no increased resistance to others first-line anti-tuberculosis drugs. Resazurin microtiter assay plate testing of Mycobacterium smegmatis susceptibility to drugs is a rapid, simple, and inexpensive method and could decrease color background of drugs by detecting fluorescence. It will be benefit for high-throughout screening of drugs of anti-isoniazid-resistant Mycobacteria.
