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.

ABSTRACT

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