title: Validation of qPCR Methods for the Detection of Mycobacterium in New World Animal Reservoirs.
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.
authors.affiliation: School of Human Evolution and Social Change, Arizona State University, Tempe, Arizona, United States of America.
authors.forename: Genevieve
authors.initials: G
authors.lastname: Housman
authors.affiliation: School of Human Evolution and Social Change, Arizona State University, Tempe, Arizona, United States of America.|Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil.
authors.forename: Joanna
authors.initials: J
authors.lastname: Malukiewicz
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil.
authors.forename: Vanner
authors.initials: V
authors.lastname: Boere
authors.affiliation: Laboratório de Ciências Ambientias, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Rio de Janeiro, Brazil.
authors.forename: Adriana D
authors.initials: AD
authors.lastname: Grativol
authors.affiliation: Centro de Conservação e Manejo de Fauna da Caatinga, Universidade Federal do Vale do São Francisco, Petrolina, Pernambuco, Brazil.
authors.forename: Luiz Cezar M
authors.initials: LC
authors.lastname: Pereira
authors.affiliation: Departamento de Biologia Animal, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil.
authors.forename: Ita de Oliveira
authors.initials: Ide O
authors.lastname: Silva
authors.affiliation: Laboratório de Ciências Ambientias, Centro de Biociências e Biotecnologia, Universidade Estadual do Norte Fluminense, Campos dos Goytacazes, Rio de Janeiro, Brazil.
authors.forename: Carlos R
authors.initials: CR
authors.lastname: Ruiz-Miranda
authors.affiliation: HHS\HRSA\HSB National Hansen's Disease Program-NIAID IAA-2646, Baton Rouge, Louisiana, United States of America.
authors.forename: Richard
authors.initials: R
authors.lastname: Truman
authors.affiliation: School of Human Evolution and Social Change, Arizona State University, Tempe, Arizona, United States of America.
authors.forename: Anne C
authors.initials: AC
authors.lastname: Stone
mesh_terms: D000818:Animals; D002144:Callithrix; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004197:Disease Reservoirs / Q000382:microbiology / Q000662:veterinary*; D008403:Mass Screening / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009164:Mycobacterium Infections / Q000382:microbiology / Q000662:veterinary*; D009166:Mycobacterium leprae / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D009169:Mycobacterium tuberculosis / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit