biostudies-literatureGoosen WJEuropean & Developing Countries Clinical Trials Partnership (EDCTP)South African Medical Research CouncilEuropean CommissionNational Research FoundationWellcome Trust100702https://www.ebi.ac.uk/biostudies/studies/S-EPMC10937233biostudies-literatureUnknown18This study investigated the presence of <i>Mycobacterium bovis</i> (<i>M. bovis</i>) DNA in archived human sputum samples previously collected from residents who reside adjacent to the <i>M. bovis</i>-endemic Hluhluwe-iMfolozi wildlife park, South Africa (SA). Sixty-eight sputum samples were GeneXpert MTB/RIF Ultra-positive for <i>M. tuberculosis</i> complex (MTBC) DNA but culture negative for <i>M. tuberculosis</i>. Amplification and Sanger sequencing of <i>hsp65</i> and <i>rpoB</i> genes from DNA extracted from stored heat-inactivated sputum samples confirmed the presence of detectable amounts of MTBC from 20 out of the 68 sputum samples. Region of difference PCR, spoligotyping and <i>gyrB</i> long-read amplicon deep sequencing identified <i>M. bovis</i> (<i>n</i> = 10) and <i>M. tubercuOne health (Amsterdam, Netherlands)Identification and molecular characterization of <i>Mycobacterium bovis</i> DNA in GeneXpert® MTB/RIF ultra-positive, culture-negative sputum from a rural community in South Africa.PMC109372331022002201433/Z/16/A10110317186949222941/Z/21/ZHanekom WAVukuzazi TeamWong EBSmit TMiller MAGhielmetti GStreicher EMWarren RMMoodley SGoosen WJMoosa YZulu TKleynhans LKerr TJfalseIdentification and molecular characterization of <i>Mycobacterium bovis</i> DNA in GeneXpert® MTB/RIF ultra-positive, culture-negative sputum from a rural community in South Africa.This study investigated the presence of <i>Mycobacterium bovis</i> (<i>M. bovis</i>) DNA in archived human sputum samples previously collected from residents who reside adjacent to the <i>M. bovis</i>-endemic Hluhluwe-iMfolozi wildlife park, South Africa (SA). Sixty-eight sputum samples were GeneXpert MTB/RIF Ultra-positive for <i>M. tuberculosis</i> complex (MTBC) DNA but culture negative for <i>M. tuberculosis</i>. Amplification and Sanger sequencing of <i>hsp65</i> and <i>rpoB</i> genes from DNA extracted from stored heat-inactivated sputum samples confirmed the presence of detectable amounts of MTBC from 20 out of the 68 sputum samples. Region of difference PCR, spoligotyping and <i>gyrB</i> long-read amplicon deep sequencing identified <i>M. bovis</i> (<i>n</i> = 10) and <i>M. tubercu2024-01-01T00:00:00Z2024 Jun2026-07-15T00:44:56.089Z2026-06-27T03:06:16.885ZS-EPMC109372333848772910.1016/j.onehlt.2024.100702