Designing and development of NASBA for detection of EF-Tu mRNA of Mycobacterium tuberculosis

Accurate and early diagnosis of tuberculosis id important for its effective management. Because of the slow growth rate of Mycobacterium tuberculosis in culture and sever linitations of conventional methods (such as microscopic examination)applied in Iran, perceived the need of rapid, sensitive and accurate techniques for indentification of Mycobacterium tuberculosis. During las decade several molecular methods for detection of tuberculosis (TB), have been developed. persistence of DNA, long after conversion of smears and cultures to negative, because this nucleic acid target unsuitable for monitoring therapeutic efficacy of tuberculosis. Since, RNA especially mRNA has a generally much shorter half-life than DNA, its detection may be useful for the assessment of viability of bacteria so, techniques that utilize mRNA as a template are more suitable for detection of Mycobacterium tuberculosis. We have developed Nucleic acid sequenced-based amplication (NASBA), as an isothemal amplification technique for identification of Mycobacterium tuberculosis EF-Tu mRNA. Total RNA was extracted from pure culture of Mycobacterium tubercuosis H37Rv and NASBA was carried out to detect EF-Tu mRNA. The amplicons were detected by Gel-electrophoresis in RNase-free format. This study confirmed that NASBA can be a rapid and sensitive technique for detection ofMycobacterium tuberculosis EF-Tu mRNA.

کلیدواژه ها:

NASBA ، EF-Tu mRNA ، Mycobacterium tuberculosis ، viability