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Title: USE OF A NOVEL VIRUS INACTIVATION METHOD FOR A MULTI-CENTER AVIAN INFLUENZA REAL-TIME RT-PCR PROFIEICNEY

Author
item Spackman, Erica
item Suarez, David

Submitted to: Journal of Veterinary Diagnostic Investigation
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/7/2004
Publication Date: 2/1/2005
Citation: Spackman, E., Suarez, D.L. Use of a novel virus inactivation method for a multi-center avian influenza real-time rt-pcr profieicney. Journal of Veterinary Diagnostic Investigation, 17:76-80 (2005).

Interpretive Summary: Proficiency assessments are an essential part of quality assurance for diagnostic laboratories and have been implemented for many assay types including PCR based tests, which detect the genetic material of disease causing agents. 'Real-time RT-PCR' is a relatively new technology for the detection of the genetic material of disease causing agents, which due to its many advantages (very rapid, sensitive and specific) is being developed and implemented for the detection of many disease causing agents from many species. The robustness of this technology has not been evaluated due to the relative newness of the technology. Due to the high economic importance of avian influenza or 'bird flu' one of the first real-time RT-PCR tests in veterinary diagnostics was developed and implemented for this agent. This is a standardized test used by USDA and the national animal health network laboratories and has been used during an outbreak of bird flu in the US. This study is the first assessment of the performance of this technology in the field. The test performed with a high level of accuracy and precision among 12 laboratories. The high level of performance of this test in this evaluation show that this is a an appropriate method with high utility.

Technical Abstract: A real-time RT-PCR test for type A influenza was shown to be highly reproducible by proficiency testing of 12 laboratories using a novel sample medium for inactivating and stabilizing the virus. Variation in cycle threshold values among 35 data-sets and 490 samples was minimal (CV=5.19%) and sample identifications were highly accurate (96.7% correct identifications) regardless of real-time PCR instrumentation.