Direct DNA Modified CTAB Preparation from Nasal Exudate in Live M. bovis Infected Cattle in Mexico Provide with a Valuable Assay Extrapolated to Humans TB Diagnostic Test

Sanchez-Garza J.J, Garcia-Roqu

Abstract

Detection, identification, and differentiation of members of the MTB complex rely on specificity, sensitivity, and accuracy of the methods that have been developed since the decades of the ’90s. Despite this, still, in endemic areas of developing countries tuberculin field test as well as conventional techniques (histopathology and bacteriology) are performed due primarily to the costs and availability. Therefore, it is an urgent need to have a routine assay to boost field test (false positive and negative tests) in live cows while avoiding the unnecessary sacrifice of animals. To this end, in the present work, we designed a dual experimental strategy that can be used as a routine assay for the M. bovis or M. tuberculosis detection through PCR mediated amplification of RD’s. DNA can be prepared from fast-growing colonies (7 to 8 days) or from homogenized tissue, nasal exudate and purification mediated cetyl-trimethylammonium bromide (CTAB) cationic buffer. The method was extraplated to positive TB positive nasal/oral human exudate.with similar results. Collectively these findings indicate that this strategy represent a valuable tool for TBb epidemiological survey and research.

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