Use of reverse transcriptase polymerase chain reaction (RT-PCR) in molecular screening of Newcastle disease virus in poultry and free-living bird populations

• Published in: Tropical animal health and production 2013-02, Vol.45 (2), p.569-576
• Main Authors: de Oliveira Torres Carrasco, Adriano, Rodrigues, Juliana Nogueira Martins, Seki, Meire Christina, de Moraes, Fabricio Edgar, Silva, Jaqueline Raymondi, Durigon, Edison Luis, Pinto, Aramis Augusto
• Format: Article
• Language: English
• Subjects: Animal models; Animals; Aves; Biomedical and Life Sciences; Bird populations; Birds; Chickens; Columbidae; DNA Primers - genetics; Feces; Feces - virology; Life Sciences; Molecular modelling; Newcastle disease; Newcastle Disease - diagnosis; Newcastle Disease - virology; Newcastle disease virus; Newcastle disease virus - genetics; Newcastle disease virus - isolation & purification; Pathogenicity; Pathogens; Polymerase chain reaction; Poultry; Poultry Diseases - diagnosis; Poultry Diseases - virology; Primers; Regular Articles; Reverse Transcriptase Polymerase Chain Reaction - methods; Reverse Transcriptase Polymerase Chain Reaction - veterinary; RNA-directed DNA polymerase; Sensitivity and Specificity; Stains; Vaccination; Veterinary Medicine/Veterinary Science; Virus Shedding; Zoology
• ISSN: 0049-4747; 1573-7438
• DOI: 10.1007/s11250-012-0261-7

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.