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Avian Influenza Virus RNA Extraction

  • Erica SpackmanEmail author
  • Scott A. Lee
Protocol
  • 128 Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 2123)

Abstract

The efficient extraction and purification of viral RNA is critical for downstream molecular applications such as the sensitive and specific detection of virus in clinical samples, virus gene cloning and expression, gene sequencing, or quantification of avian influenza (AI) virus by molecular methods from experimentally infected birds. Samples can generally be divided into two types: enriched (e.g., virus stocks) and non-enriched (e.g., clinical). Clinical samples, which may be tissues or swab material, are the most difficult to process due to the complex sample composition and possibly low virus titers. In this chapter, two well-established procedures for the extraction of AI virus RNA from common clinical specimen types and enriched virus stocks will be presented.

Key words

RNA extraction Influenza nucleic acid Specimen processing Swab material Avian influenza 

References

  1. 1.
    Das A, Spackman E, Pantin-Jackwood MJ, Suarez DL (2009) Removal of real-time reverse transcription polymerase chain reaction (RT-PCR) inhibitors associated with cloacal swab samples and tissues for improved diagnosis of Avian influenza virus by RT-PCR. J Vet Diagn Investig 21(6):771–778CrossRefGoogle Scholar
  2. 2.
    Spackman E, Senne DA, Myers TJ, Bulaga LL, Garber LP, Perdue ML, Lohman K, Daum LT, Suarez DL (2002) Development of a real-time reverse transcriptase PCR assay for type A influenza virus and the avian H5 and H7 hemagglutinin subtypes. J Clin Microbiol 40(9):3256–3260CrossRefGoogle Scholar
  3. 3.
    Spackman E, Suarez DL (2005) Use of a novel virus inactivation method for a multicenter avian influenza real-time reverse transcriptase-polymerase chain reaction proficiency study. J Vet Diagn Investig 17(1):76–80CrossRefGoogle Scholar
  4. 4.
    Erickson GA, Brugh M, Beard CW (1978) Newcastle disease and avian influenza virus stability under simulated shipping conditions. Twenyfirst annual meeting of american association of veterinary laboratory diagnosticians. 309–318Google Scholar
  5. 5.
    Das A, Spackman E, Senne D, Pedersen J, Suarez DL (2006) Development of an internal positive control for rapid diagnosis of avian influenza virus infections by real-time reverse transcription-PCR with lyophilized reagents. J Clin Microbiol 44(9):3065–3073CrossRefGoogle Scholar

Copyright information

© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply. 2020

Authors and Affiliations

  1. 1.Exotic and Emerging Avian Viral Diseases Unit, US National Poultry Research CenterUS Department of Agriculture, Agricultural Research ServiceAthensUSA

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