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Journal of Veterinary Diagnostic Investigation Vol. 20 Issue 5, 612-616
Copyright © 2008 by the American Association of Veterinary Laboratory Diagnosticians
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Brief Research Reports

Analytical validation of a real-time reverse transcription polymerase chain reaction test for Pan-American lineage H7 subtype Avian influenza viruses

Erica Spackman1, Hon S. Ip, David L. Suarez, Richard D. Slemons and David E. Stallknecht

Correspondence: 1Corresponding Author: Erica Spackman, SEPRL, USDA, ARS, 934 College Station Road, Athens, GA 30605. erica.spackman{at}ars.usda.gov

A real-time reverse transcription polymerase chain reaction test for the identification of the H7 subtype in North American avian influenza viruses AIV was first reported in 2002; however, recent (AIVs) surveillance efforts in wild birds and H7 outbreaks in poultry demonstrated that the 2002 test did not detect all H7 AIVs present in North and South America. Therefore, a new test, the 2008 Pan-American H7 test, was developed by using recently available H7 nucleotide sequences. The analytical specificity of the new assay was characterized with an RNA panel composed of 19 H7 viruses from around the world and RNA from all hemagglutinin subtypes except H16. Specificity for North and South American lineage H7 viruses was observed. Assay limits of detection were determined to be between 103 and 104 gene copies per reaction with in vitro transcribed RNA, and 100.0 and 100.8 50% egg infectious doses per reaction. The 2008 Pan-American H7 test also was shown to perform similarly to the 2002 test with specimens from chickens experimentally exposed to A/Chicken/BritishColumbia/314514-2/04 H7N3 highly pathogenic AIV. Furthermore, the 2008 test was able to detect 100% (n = 27) of the H7 AI, V isolates recovered from North American wild birds in a 2006–2007 sample set, (none of which were detected by the 2002 H7 test).

Key Words: Avian influenza • H7 hemagglutinin subtype • poultry • real-time reverse transcription polymerase chain reaction • wild bird disease






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