Indian Journal of Comparative Microbiology, Immunology and Infectious Diseases
Year : 2003, Volume : 24, Issue : 2
First page : ( 167) Last page : ( 171)
Print ISSN : 0970-9320. Online ISSN : 0974-0147.

Detection of Mycobacterium tuberculosis complex organisms in clinical samples of cattle by pcr and dna probe methods

Sreedevi B.¹, Assistant Professor, Krishnappa G.

Department of Veterinary Microbiology, College of Veterinary Science University of Agricultural Sciences, Hebbal, Bangalore–560 024 (Karnataka)

¹Department of Veterinary Epidemiology and Preventive Medicine, College of Veterinary Science, Acharya N.G.Ranga Agricultural University, Tirupathi–517 502, Andhra Pradesh, India

Received:  30  September,  2003.

Abstract

In the present study, the PCR technique was standardized using primers for IS6110 element specific to M. tuberculosis complex group of organisms and amplification was observed with M. bovis M. tuberculosis and M. bovis BCG strains while the remaining controls, M. phlei M. avium andM. paratuberculosis were negative. The purified PCR product labelled with digoxigenin in the hybridization studies gave positive signal with M. tuberculosis complex strains only, where as negative controls including M. paratuberculosisM. phlei and M. avium did not reveal any hybridization. The dot blot hybridization with different mycobacterial DNAs further confirmed the results of PCR. The efficacy of PCR and dot blot hybridization for the detection of M. tuberculosis complex group of organisms in the clinical samples were found to be 60 to 75 per cent with blood samples, 40 to 75 per cent with nasal swabs, 15 per cent with milk samples and none of the semen samples collected from tuberculin test positive cattle were positive by PCR and dot blot assay. All the samples collected from tuberculintest negative animals were also negative by both PCR and dot blot assay.

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