Detection of Mycobacterium tuberculosis Complex in Lung Specimen of Slaughtered Cattle and Goats by a DNA Based multiplex Polymerase Chain Reaction and Ziehl-Neelsen Methods In Jos, Nigeria

Abstract

Aims: Determine percentage positive by the Ziehl-Neelsen (ZN) stain; detect 245 base pair fragment of the IS6110 gene for Mycobacterium tuberculosis complex using INS1 and INS2 primers and 500 base pair fragment of the RvD1Rv2031c gene for M. bovis using the JB21 and JB22 primers by a multiplex PCR (M-PCR);compare number of positive samples detected by ZN stain and M–PCR; determine prevalence rates of Mycobacterium tuberculosis complex between the two animal species studied and estimate the rates of detecting agents of tuberculosis using Ziehl-Neelsen (ZN) technique and a Multiplex-Polymerase Chain Reaction (M-PCR) in lung samples of slaughtered cattle and goats in the study area. Place and Duration of Study: Samples were analyzed at the Central Diagnostic Laboratory and Molecular Biology Departments of the National Veterinary Research Institute Vom. This work was carried out between July-October 2010. Study Design: Experimental. Methodology: Our PCR amplified the 245 base pair (bp) fragment which is specific for this group of Mycobacterium while ZN exploited the acid-fast nature of the organisms. We examined one hundred lung samples, of cattle and goats, fifty for each. Results: Out of the lung samples from cattle, 15 (30%) were positive by ZN while 9 (18%) were positive by PCR. Among the ZN negative samples from cattle, one was positive by M-PCR. All the 50 samples from goats were negative by the two diagnostic tools used in this study. Results obtained in this study showed 0% TB infection in goats and 18% in cattle. Conclusion: The study showed a high infection rate of tuberculosis among cattle sampled in the area of study, as such, preventive and curative measures have to be stepped up in controlling the zoonosis.