Impregnation and storage of Newcastle disease virus on to filter papers and detection of viral RNA by a single tube RT-PCR assay

Abstract

Suitability of storing infected allantoic fluid (AF) and cell culture supernatants (CCS) with strain I-2 of Newcastle disease virus (NDV) on Whatman filter papers at room temperature (22-25°C) and 37°C was determined. RNA was extracted from filter papers or liquid aliquots and subjected to reverse transcriptase-polymerase chain reactions (RT-PCR). The results showed that filter papers soaked with NDV infected AF or CCS stored at 37°C yielded amplicons with intensity similar to that kept at room temperature for up to 150 days. The study demonstrates that NDV infected samples can be soaked onto filter papers, stored and subsequently detected by RT PCR. This method might be safely used for storage and transportation of NDV samples to the designated laboratories for molecular studies without the need for cooling.

Keywords: Allantoic fluid, chicken embryo fibroblast, Newcastle diseases virus, polymerase chain reaction, RNA storage onto filter paper, strain I-2