Carrasco Adriano de Oliveira Torres, Rodrigues Juliana Nogueira Martins, Seki Meire Christina, de Moraes Fabricio Edgar, Silva Jaqueline Raymondi, Durigon Edison Luis, Pinto Aramis Augusto
Departamento de Medicina Veterinária, Universidade Estadual do Centro-Oeste, UNICENTRO, Guarapuava, Parana, Brazil.
Trop Anim Health Prod. 2013 Feb;45(2):569-76. doi: 10.1007/s11250-012-0261-7. Epub 2012 Sep 15.
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.
本研究的目的是评估一种简单的逆转录聚合酶链反应（RT-PCR）分子方法，以根据新城疫病毒毒株的致病性对其进行区分，以便将其用于家禽和自由生活鸟类群体中新城疫病毒的分子筛查。开发了特异性引物来区分LaSota（LS，疫苗株）和圣若昂·多梅里蒂（SJM）毒株（高致病性毒株）。通过对鸡和鸽子进行实验性接种/感染以进行体内研究，来确定粪便中的病毒排出情况。通过本研究中使用的实验模型对引物（SJM和LS）的敏感性和特异性进行验证，以及通过BLAST对引物进行分子分析所获得的结果，使得研究结果具有普遍性。开发能够区分新城疫病毒毒株致病性水平的引物非常重要，主要是在尚未将实时RT-PCR用作常规检测的国家。这些引物能够确定病原体的存在，并根据其致病性进行区分。
