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

• Published in: PLOS Neglected Tropical Diseases
• Main Authors: Genevieve Housman, Joanna Malukiewicz, Vanner Boere, Adriana D Grativol, Luiz Cezar M Pereira, Ita de Oliveira Silva, Carlos R Ruiz-Miranda, Richard Truman, Anne C Stone
• Format: Article in Journal/Newspaper
• Language: English
• Published: Public Library of Science (PLoS) 2015
• Subjects: Arctic medicine. Tropical medicine; RC955-962; Public aspects of medicine; RA1-1270; Arctic
• Online Access: https://doi.org/10.1371/journal.pntd.0004198; https://doaj.org/article/01a7431be01b481cb4cc7864c3b3e04e

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.