Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs

Genevieve Housman, Joanna Malukiewicz, Vanner Boere, Adriana D. Grativol, Luiz Cezar M Pereira, Ita de Oliveira e Silva, Carlos R. Ruiz-Miranda, Richard Truman, Anne Stone

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.

Original languageEnglish (US)
Article numbere0004198
JournalPLoS Neglected Tropical Diseases
Volume9
Issue number11
DOIs
StatePublished - Nov 16 2015

ASJC Scopus subject areas

  • Public Health, Environmental and Occupational Health
  • Infectious Diseases

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    Housman, G., Malukiewicz, J., Boere, V., Grativol, A. D., Pereira, L. C. M., Silva, I. D. O. E., Ruiz-Miranda, C. R., Truman, R., & Stone, A. (2015). Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs. PLoS Neglected Tropical Diseases, 9(11), [e0004198]. https://doi.org/10.1371/journal.pntd.0004198