Measurement of Avian Cytokines with Real-Time RT-PCR Following Infection with the Avian Influenza Virus

Functional and molecular techniques have both been employed to measure the production of cytokines following influenza infection. Historically, the use of functional or antibody-based techniques was employed in mammalian immunology. In avian immunology, only a few commercial antibodies are available to measure avian cytokines. However, the determination of the genomic sequence of Gallus gallus species has made it possible to measure cytokine induction without monoclonal antibody- or functional-based tests, but rather based on molecular techniques. Although these tests do not measure functionally expressed cytokines, the lack of reagents to identify and quantify avian cytokines makes them critical to extend any measure of cytokine response. Measurement of cytokine induction, based on the design of primers and probes for RT-PCR or real-time RT-PCR for the cytokine mRNA, has become one of the more recent technologies reported to measure avian cytokines. It is important to note that small nucleotide polymorphisms between different lines of birds may result in substandard results when using published primer and probe sequences. This requires empirical testing to ensure adequate results.