African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Full Length Research Paper

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

P. N. Wambura
  • P. N. Wambura
  • School of Veterinary Science, Faculty of Natural Resources, Agriculture and Veterinary Science, University of Queensland, Brisbane, QLD 4072, Australia.
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H. Khazali
  • H. Khazali
  • Shahid Beheshti University, Tehran, Iran.
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A. Parhiskar
  • A. Parhiskar
  • Shahid Beheshti University, Tehran, Iran.
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K. Jafari
  • K. Jafari
  • Shahid Beheshti University, Tehran, Iran.
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  •  Accepted: 21 October 2005
  •  Published: 02 January 2006

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.

 

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