Development of a Highly Sensitive Screen for Influenza A in Guano and Its Application in the Search for Ancient RNA Preserved Under Antarctic Adelie Penguin Colonies

• Published in: Avian Diseases
• Main Authors: L. C. Briggs, R. M. Ashton, P. Metcalf
• Format: Text
• Language: English
• Published: American Association of Avian Pathologists 2003
• Subjects: Antarctic; Guano; Adelie penguin; Antarc*; Pygoscelis adeliae
• Online Access: https://doi.org/10.1637/0005-2086-47.s3.1200

We have developed a reverse transcriptase polymerase chain reaction (RT-PCR)-based assay to detect influenza A in guano samples as part of our program to investigate ancient viral RNA from under Antarctic Adelie penguin (Pygoscelis adeliae) colonies. Of five extraction protocols tested (RNeasy, GTC TRIZOL, GTC Silica, Rnaid, and AGPC), AGPC proved to be the most consistent and sensitive to low concentrations of influenza, but further purification with commercial viral nucleic acid spin filter system was still required to remove remaining PCR inhibitors. RT-PCR was then performed on the eluent and 650 bases of the M1 gene were amplified. The assay was found to be able to detect as little as 100 μl of 0.1 hemagglutination units (HU)/ml influenza.