Ormeño-Vásquez P.1, Tataje-Lavanda L.1, Huamán-Gutiérrez K.1, Calderón-Mayo K.1, Longa-Bobadilla V.1, Montalván Avalos A.1, Huaccachi-Gónzales E.1, Criollo-Orozco M.1 , Bendezú J.1 , Fernández-Díaz M.1.
1FARVET, Research and Development Laboratories, Ica, Peru.
The Newcastle virus is the causative agent of Newcastle disease which is highly worldwide distribution. The best prevention measure is vaccination, therefore, a technique is required for its titration that is precise, fast, sensitive and specific.
The objective of the study was to develop a molecular platform for the quantification of NDV in an SPF embryonated egg culture system. Four pairs of primers were evaluated, directed at nucleocapsid (NP), matrix (M), fusion (F) and RNA-dependent RNA polymerase (L) genes, using two-step RT-PCRc (reverse transcription polymerase chain reaction) for the detection of NDV. Subsequently, it was taken to RT-qPCR (real-time reverse transcription polymerase chain reaction) for the quantification of NDV. Using these techniques, it was determined that the primers for the M gene were the most suitable for the development of both methods. Sensitivity tests showed that the RT-qPCR was 10 times more sensitive (116 genomic copies/μl) than the RT-PCRc and the primers were specific. Due to its sensitivity and specificity, this platform is proposed for the quantification of the NDV vaccine in embryonated egg system, as a method of titration.
To Access the original article, check the following link: https://www.ncbi.nlm.nih.gov/pubmed/28301239