Quantitative Polymerase Chain Reaction (PCR) for Detection of Aquatic Animal Pathogens in a Diagnostic Laboratory Setting |
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| Authors: | Maureen K. Purcell Rodman G. Getchell Carol A. McClure Kyle A. Garver |
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| Affiliation: | 1. U.S. Geological Survey, Western Fisheries Research Center , 6505 North East 65th Street, Seattle , Washington , 98115 , USA;2. Department of Microbiology and Immunology , College of Veterinary Medicine, Cornell University , Upper Tower Road, Ithaca , New York , 14853 , USA;3. Center for Aquatic Health Sciences, University of Prince Edward Island , 550 University Avenue, Charlottetown, Prince Edward Island C1A 4P3, Canada;4. AquaEpi Research , 300 Heartz Road, Charlottetown, Prince Edward Island C1A 7J7, Canada;5. Fisheries and Oceans Canada, Aquatic Animal Health, Pacific Biological Station , 3190 Hammond Bay Road, Nanaimo, British Columbia V9T 6N7, Canada |
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| Abstract: | Abstract Real-time, or quantitative, polymerase chain reaction (qPCR) is quickly supplanting other molecular methods for detecting the nucleic acids of human and other animal pathogens owing to the speed and robustness of the technology. As the aquatic animal health community moves toward implementing national diagnostic testing schemes, it will need to evaluate how qPCR technology should be employed. This review outlines the basic principles of qPCR technology, considerations for assay development, standards and controls, assay performance, diagnostic validation, implementation in the diagnostic laboratory, and quality assurance and control measures. These factors are fundamental for ensuring the validity of qPCR assay results obtained in the diagnostic laboratory setting. Received March 30, 2011; accepted May 6, 2011 |
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