Submitted to: Animal Influenza Virus
Publication Type: Book / Chapter
Publication Acceptance Date: 5/1/2019
Publication Date: 3/27/2020
Citation: Spackman, E. 2020. Avian influenza virus detection and quantitation by real-time RT-PCR. In: Spackman, E., Editor. Animal Influenza Virus Methods and Protocols. 3rd Edition. Methods in Molecular Biology, volume 2123. New York, NY: Humana. p. 137-148. https://doi.org/10.1007/978-1-0716-0346-8_11.
Technical Abstract: Real-time RT-PCR (rRT-PCR) has been used for avian influenza virus (AIV) detection since the early 2000’s for routine surveillance, outbreaks assessment and for research. Some of the advantages of rRT-PCR are: high sensitivity, high specificity, rapid time-to-result, scalability, cost, and its inherently quantitative nature. Furthermore, rRT-PCR can be used with numerous sample types, is less expensive than virus isolation in chicken embryos, and since infectious virus is inactivated early during processing, biosafety and bio-security are also easier to maintain. However, the high genetic variability of AIV may decrease sensitivity and increases the chances of a false negative result. This chapter will provide an overview of the USDA validated rRT-PCR procedure for the detection of type A influenza.