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Title: Validation of a real-time reverse transcriptase-PCR assay for the detection of H7 avian influenza virus

Author
item PEDERSEN, JANICE - Animal And Plant Health Inspection Service (APHIS)
item KILLIAN, MARY LEA - Animal And Plant Health Inspection Service (APHIS)
item HINES, NICHOLE - Animal And Plant Health Inspection Service (APHIS)
item SENNE, DENNIS - Animal And Plant Health Inspection Service (APHIS)
item PANIGRAHY, BRUNDABAN - Animal And Plant Health Inspection Service (APHIS)
item IP, HON - US Department Of Interior
item Spackman, Erica

Submitted to: Avian Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/15/2009
Publication Date: 7/1/2010
Citation: Pedersen, J., Killian, M., Hines, N., Senne, D., Panigrahy, B., Ip, H., Spackman, E. 2010. Validation of a real-time reverse transcriptase-PCR assay for the detection of H7 avian influenza virus. Avian Diseases. 54:639-643.

Interpretive Summary: The most important subtypes of influenza for poultry are H5 and H7. Currently there are several methods of identifying an influenza virus H subtype and one of the most common detects the H7 genetic material, ribonucleic acid (RNA). Since influenza can mutate very rapidly periodic updates for the test are necessary to identify new strains. This study evaluates the updated genetic test for H7 and provides data which demonstrates that it performs better than the previous version of the test and therefore has been adopted as the current official United States Department of Agriculture (USDA) test.

Technical Abstract: A subtype specific H7 real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay developed by the Southeast Poultry Research Laboratory (SEPRL) for the detection of H7 in North and South American wild aquatic birds and poultry was validated as a collaborative effort by the SEPRL and National Veterinary Services Laboratories (NVSL). The 2008 H7 assay will detect 101 EID50 per reaction or 103 - 10 4 copies of transcribed RNA. Diagnostic sensitivity and specificity was estimated to be 97.5% and 82.4%, respectively and the assay was shown to be specific for H7 AI when tested with >270 viruses isolated from wild birds and poultry. The procedure was validated as the United States Department of Agriculture (USDA) official procedure for the detection of H7 AI and has replaced the previous 2002 assay which was shown not to detect the lineage of H7 currently circulating in wild birds.