Easy, fast, point of care nucleic acid based detection of pulmonary tuberculosis

Koshinsky, Heather

Abstract

The proposed research is to determine the feasibility of developing a novel, simple, rapid method of detecting the nucleic acid (NA) from Mycobacterium tuberculosis cell in sputum, for detection of pulmonary tuberculosis (TB). This method would be based on Investigen's novel and elegant """"""""smartDNA"""""""" technology. This system utilizes a peptide nucleic acid (PNA) probe cocktail in conjunction with a dye and light activation to produce a rapid color change when probes hybridize to complementary NA sequences. This color change can be visualized by the naked eye or detected by an absorbance or fluorescence reader. The four aims of feasibility are 1) evaluate the use synthetic sputum and M. microti as a surrogate for human sputum and M. tuberculosis; 2) develop a sputum processing protocol that is compatible with smartDNA-based detection of TB; 3) determine specificity and sensitivity of the entire sputum processing- smartDNA TB detection system in the detection of M. tuberculosis NA and 4) validate that our simple, rapid lysis method releases NA from M. tuberculosis. With this validation of the feasibility of developing a complete smartDNA TB detection system, we would move to Phase II objectives and test the system with normal human sputum samples spiked with M. tuberculosis and with clinical samples. The intention is to create a commercial diagnostic assay to detect TB in patients. Such a diagnostic could be used in peripheral locations, away from a central lab. This is greatly needed and would help curb the spread of TB by offering more rapid treatment. The proposed research would use a surrogate system to examine the feasibility of developing a method for the rapid detection of M. tuberculosis out of sputum. Findings from this study would be very useful in developing a novel commercial test for patients with active TB. ? ? ?