Mutation screening in genes related to drug resistance in Mycobacterium tuberculosis isolates by real time PCR High Resolution Melting Curve

• Grant number: 09/14329-9
• Support Opportunities: Regular Research Grants
• Start date: January 01, 2010
• End date: December 31, 2011
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Mario Hiroyuki Hirata
• Grantee: Mario Hiroyuki Hirata
• Host Institution: Faculdade de Ciências Farmacêuticas (FCF). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The mycobacterial resistance to the drugs used in the treatment of tuberculosis has encouraged the research of new drugs, their action mechanism and the development of fast and affordable techniques to detect the bacterial drug resistance. The aim of this study is to study a new system to detect mycobacterial mutations related to resistance to ethambutol, isoniazid and rifampicin by using the Real Time PCR -High resolution Melting Curve. The genes KatG, EmbB, rpoB and the promoter region of InhA and AhpC will be studied. A total of 80 clinical isolates of mycobacterium tuberculosis from the laboratory library, which have the fenotipic drug resistance profile established will be evaluated. The results obtained by qPCR-HRM will be compared with data obtained from DNA sequecing. The new system might present a low cost, high sensibility and reproducibility when compared with other techniques such as PCR-SSCP and DNA sequencing. (AU)