Detection of New Viruses by VIDISCA

Virus Discovery Based on cDNA-Amplified Fragment Length Polymorphism
  • Krzysztof Pyrc
  • Maarten F. Jebbink
  • Ben Berkhout
  • Lia van der Hoek
Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 454)

Abstract

Virus discovery based on cDNA-AFLP (amplified fragment length polymorphism) (VIDISCA) is a novel approach that provides a fast and effective tool for amplification of unknown genomes, e.g., of human pathogenic viruses. The VIDISCA method is based on double restriction enzyme processing of a target sequence and ligation of oligonucleotide adaptors that subsequently serve as priming sites for amplification. As the method is based on the common presence of restriction sites, it results in the generation of reproducible, species-specific amplification patterns. The method allows amplification and identification of viral RNA/DNA, with a lower cutoff value of 105 copies/ml for DNA viruses and 106 copies/ml for the RNA viruses. Previously, we described the identification of a novel human coronavirus, HCoV-NL63, with the use of the VIDISCA method.

Key Words

VIDISCA virus discovery detection diagnosis cDNA-AFLP amplification RT-PCR 
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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Krzysztof Pyrc
    • 1
  • Maarten F. Jebbink
    • 1
  • Ben Berkhout
    • 1
  • Lia van der Hoek
    • 1
  1. 1.Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical CenterUniversity of AmsterdamAmsterdamThe Netherlands

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