Quantitative polymerase chain reaction (PCR) for detection of aquatic animal pathogens in a diagnostic laboratory setting.
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.
U.S. Geological Survey, Western Fisheries Research Center 6505 North East 65th Street, Seattle, Washington 98115, USA. mpurcell@usgs.gov
Maureen K
MK
Purcell
Rodman G
RG
Getchell
Carol A
CA
McClure
Kyle A
KA
Garver
D000820:Animal Diseases / Q000175:diagnosis*; D000818:Animals; D007753:Laboratories; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity