Wynand Goosen Poster 2024

Wynand Johan Goosen

Dr Wynand Goosen

Stellenbosch University, South Africa

Identification and Molecular Characterization of Mycobacterium bovis DNA in GeneXpert® MTB/RIF Ultra-Positive, Culture-Negative Sputum Samples from a Rural Community in South Africa


Poster Abstract

This study investigated the presence of Mycobacterium bovis (M. bovis) DNA in human sputum samples collected from residents who reside adjacent to the M. bovis-endemic HluhluweiMfolozi wildlife park, South Africa (SA). Sixty-eight sputum samples were GeneXpert MTB/RIF Ultra-positive for M. tuberculosis complex (MTBC) DNA but culture negative for M. tuberculosis. Amplification and Sanger sequencing of hsp65 and rpoB genes from residual frozen sputum confirmed the presence of detectable amounts of MTBC in DNA extracted from 20 out of the 68 sputum samples. Region of difference PCR, spoligotyping and gyrB long-read amplicon deep sequencing together identified M. bovis (n = 10) from the 20 samples from which MTBC DNA could be detected. Notably, M. bovis spoligotypes SB0130 and SB1474 were identified in 4 samples, with SB0130 previously identified in local cattle and wildlife and SB1474 exclusively in African buffaloes in the adjacent park. M. bovis DNA in sputum, from people living near the park, underscores zoonotic transmission potential in SA. Identification of spoligotypes specifically associated with wildlife only and spoligotypes found in livestock as well as wildlife, highlights the complexity of TB epidemiology at wildlife-livestock-human interfaces. These findings emphasize the need for integrated surveillance and control strategies to curb potential spillover and for the consideration of human M. bovis infection in SA patients with positive Ultra results. 



Wynand Goosen is fascinated and devoted to understanding and improving the genomic surveillance of zoonotic tuberculosis (TB) in people, livestock, wildlife, and their environments with a strong global health focus. Growing up in rural Eastern-Cape, he was always surrounded by various disease management strategies in livestock and wildlife due to their frequent interactions with people. This inspired him to become an independent scientific leader in zoonotic infectious diseases.

His current research focus includes:

  1. Developing rapid in-field host- and TB antigen specific cell-mediated immunological cytokine release assays.
  2. Performing rapid DNA and RNA sequencing (including PCR amplicons) using various next generation sequencing (NGS) platforms, including Oxford Nanopore Technologies MinION mk1c device.
  3. Gene expression assay development.
  4. Developing real-time quantitative PCRs for the direct detection of pathogens from various sample types and culture-independent speciation through targeted amplicon NGS.
  5. Improving BLS-3 mycobacterial culture techniques through developing cationic D-enantiomer peptide resuscitation cocktails.
  6. Application of adaptive whole-genome sequencing directly from raw specimens and cultured TB isolates using ONT newest chemistries and devices.
  7. Applying various bioinformatic pipelines to perform epidemiological transmission network analysis.