title: Use of reverse transcriptase polymerase chain reaction (RT-PCR) in molecular screening of Newcastle disease virus in poultry and free-living bird populations.
abstract: The aim of this study was to evaluate a simple molecular method of reverse transcriptase polymerase chain reaction (RT-PCR) to differentiate Newcastle disease virus strains according to their pathogenicity, in order to use it in molecular screening of Newcastle disease virus in poultry and free-living bird populations. Specific primers were developed to differentiate LaSota--LS--(vaccine strain) and Sao Joao do Meriti--SJM--strain (highly pathogenic strain). Chickens and pigeons were experimentally vaccinated/infected for an in vivo study to determine virus shedding in feces. Validation of sensitivity and specificity of the primers (SJM and LS) by experimental models used in the present study and results obtained in the molecular analysis of the primers by BLAST made it possible to generalize results. The development of primers that differentiate the level of pathogenicity of NDV stains is very important, mainly in countries where real-time RT-PCR is still not used as a routine test. These primers were able to determine the presence of the agent and to differentiate it according to its pathogenicity.
authors.affiliation: Departamento de Medicina Veterinária, Universidade Estadual do Centro-Oeste, UNICENTRO, Guarapuava, Parana, Brazil. adriano.carrasco@gmail.com
authors.forename: Adriano de Oliveira Torres
authors.initials: Ade O
authors.lastname: Carrasco
authors.forename: Juliana Nogueira Martins
authors.initials: JN
authors.lastname: Rodrigues
authors.forename: Meire Christina
authors.initials: MC
authors.lastname: Seki
authors.forename: Fabricio Edgar
authors.initials: FE
authors.lastname: de Moraes
authors.forename: Jaqueline Raymondi
authors.initials: JR
authors.lastname: Silva
authors.forename: Edison Luis
authors.initials: EL
authors.lastname: Durigon
authors.forename: Aramis Augusto
authors.initials: AA
authors.lastname: Pinto
mesh_terms: D000818:Animals; D002645:Chickens; D010856:Columbidae; D017931:DNA Primers / Q000235:genetics; D005243:Feces / Q000821:virology; D009521:Newcastle Disease / Q000175:diagnosis* / Q000821:virology; D009522:Newcastle disease virus / Q000235:genetics / Q000302:isolation & purification*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D012680:Sensitivity and Specificity; D017201:Virus Shedding
chemical_list: D017931:DNA Primers