Molecular Detection and Drug Resistance of Mycobacterium tuberculosis Complex from Cattle at a Dairy Farm in the Nkonkobe Region of South Africa: A Pilot Study

Blessing Silaigwana, Ezekiel Green and Roland N. Ndip
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Blessing Silaigwana: Department of Biochemistry and Microbiology, University of Fort Hare, P.O. Box X1314, Alice 5700, South Africa
Ezekiel Green: Department of Biochemistry and Microbiology, University of Fort Hare, P.O. Box X1314, Alice 5700, South Africa
Roland N. Ndip: Department of Biochemistry and Microbiology, University of Fort Hare, P.O. Box X1314, Alice 5700, South Africa

IJERPH, 2012, vol. 9, issue 6, 1-12

Abstract: Mycobacterium tuberculosis complex (MTBC) causes tuberculosis (TB) in humans and animals. We investigated the presence of MTBC in cattle milk and its drug resistance using polymerase chain reaction (PCR). Two hundred samples (100 mL each) were obtained from a dairy farm in the Nkonkobe region of South Africa. The samples were processed using the modified Petroff method. DNA was isolated using a Zymo Bacterial DNA kit and amplified using Seeplex ® MTB Nested ACE assay. The Genotype ® Mycobacterium tuberculosis -multidrug resistant plus (MTBDR plus ) assay was used to perform drug susceptibility and detection of mutations conferring resistance to isoniazid (INH) and rifampicin (RIF). Eleven samples tested positive for MTBC DNA using the Seeplex ® MTB Nested ACE assay. The Genotype ® MTBDR plus assay showed that 10/11 samples were resistant to both INH and RIF i.e. , multi-drug resistant (MDR). The most and least frequent rpoB mutations detected in RIF resistant samples were H526Y (9/10) and D516V (2/10) respectively. None of the INH resistant samples harbored mutations in the katG gene. However, all of them harbored the T8A mutation in the inhA gene. These results have clinical and epidemiological significance and calls for further studies and necessary actions to delineate the situation.

Keywords: Mycobacterium tuberculosis complex; unpasteurized milk; PCR; multidrug resistance; South Africa (search for similar items in EconPapers)
JEL-codes: I I1 I3 Q Q5 (search for similar items in EconPapers)
Date: 2012
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