Construction and evaluation of mycobacteriophage D29 expressing green fluorecent protein to detect Mycobacterium tuberculosis in sputum

• Grant number: 08/05076-7
• Support Opportunities: Regular Research Grants
• Start date: September 01, 2008
• End date: February 28, 2011
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• 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

Tuberculosis is a major challenge in public health to be solved. The widespread use of medicine and the abandonment of treatment by patients have selected multidrug-resistant Mycobacterium tuberculosis strains. These problems have caused difficulties to treat patients hosting these Mycobacteria. In addition, methods for diagnosis used by most public laboratories are time consuming and present low sensibility. It is very important to develop new approaches to diagnosis tuberculosis faster, with a high sensibility and low cost. The objective of this study is to set up a rapid method using a recombinant D29 phage carrying green fluorescent protein gene with a acetamidase promoter. The recombinant virus will be used as a tool for tuberculosis diagnosis. The GFP gene will be inserted in a site direct cloning using two restriction enzyme sites. Following a routine sample treatment, the purified sample will be incubated with the recombinant phage.The best time to produce detectable GFP will be evaluated in a Fluorescent ELISA automatic system. It will be a quick method with an excellent potential to reduce time and cost for TB diagnosis. (AU)