TY - JOUR
T1 - Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs
AU - Housman, Genevieve
AU - Malukiewicz, Joanna
AU - Boere, Vanner
AU - Grativol, Adriana D.
AU - Pereira, Luiz Cezar M
AU - Silva, Ita de Oliveira e
AU - Ruiz-Miranda, Carlos R.
AU - Truman, Richard
AU - Stone, Anne
N1 - Publisher Copyright:
© 2015, Public Library of Science. All Rights Reserved.
PY - 2015/11/16
Y1 - 2015/11/16
N2 - 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.
AB - 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.
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U2 - 10.1371/journal.pntd.0004198
DO - 10.1371/journal.pntd.0004198
M3 - Article
C2 - 26571269
AN - SCOPUS:84949495607
SN - 1935-2727
VL - 9
JO - PLoS Neglected Tropical Diseases
JF - PLoS Neglected Tropical Diseases
IS - 11
M1 - e0004198
ER -