title: 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.
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 resistantplus (MTBDRplus) 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(®) MTBDRplus 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.
authors: Department of Biochemistry and Microbiology, University of Fort Hare, P.O. Box X1314, Alice 5700, South Africa. silaigwana@yahoo.com
authors: Blessing
authors: B
authors: Silaigwana
authors: Ezekiel
authors: E
authors: Green
authors: Roland N
authors: RN
authors: Ndip
mesh_terms: D000818:Animals; D000904:Antibiotics, Antitubercular / Q000494:pharmacology; D000995:Antitubercular Agents / Q000494:pharmacology; D002417:Cattle; D002418:Cattle Diseases / Q000453:epidemiology* / Q000382:microbiology; D003612:Dairying; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D005260:Female; D007538:Isoniazid / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D009154:Mutation; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D010865:Pilot Projects; D015995:Prevalence; D012293:Rifampin / Q000494:pharmacology; D013019:South Africa; D014376:Tuberculosis / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*
chemical_list: D000904:Antibiotics, Antitubercular; D000995:Antitubercular Agents; D007538:Isoniazid; D012293:Rifampin