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Methods Mol Biol. 2008;436:19-26. doi: 10.1007/978-1-59745-279-3_4.

Type A influenza virus detection and quantitation by real-time RT-PCR.

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Southeast Poultry Research Laboratory, US Department of Agriculture, Agricultural Research Service, Athens, GA, USA.


Real-time RT-PCR (rRT-PCR) is a relatively new technology that has been used for avian influenza (AI) virus detection since the early 2000s for routine surveillance, during outbreaks, and for research. Some of the advantages of rRT-PCR are high sensitivity, high specificity, rapid time-to-result, scalability, cost, and 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 biosecurity are also easier to maintain. This chapter will provide an overview of the USDA-validated rRT-PCR procedure for the detection of type A influenza.

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