Development and validation of a range of endogenous controls to support the implementation of practical Taqman real‐time PCR‐based surveillance for fish diseases within aquaculture

• Published in: Journal of Fish Diseases
• Main Authors: Bland, F, McIntosh, R, Bain, N, Snow, M
• Format: Article in Journal/Newspaper
• Language: English
• Published: Wiley 2012
• Subjects: Norway; Anguilla anguilla; Atlantic salmon; European eel; Gadus morhua; Salmo salar
• Online Access: http://dx.doi.org/10.1111/j.1365-2761.2012.01363.x; https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1111%2Fj.1365-2761.2012.01363.x; https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-2761.2012.01363.x

Abstract The use of Taqman real‐time PCR‐based technology has recently become more frequent in the detection of pathogens in the aquaculture industry. This interest has necessitated the development of robust and reliable pathogen‐detection assays. The development of a range of endogenous control assays to be run alongside these diagnostic assays works to further increase confidence in the latter. This study describes the design of a range of endogenous control assays based on the elongation factor 1‐α (EF1‐α) gene specific to a range of fish species including Atlantic salmon, Salmo salar rainbow trout, Oncorhynchus mykiss; brown trout, Salmo trutta; cod, Gadus morhua; haddock, Melanogrammus aeglefinus; saithe, Pollachius virens; whiting, Merlangius merlangus; Norway pout, Trisopterus esmarkii; carp (family Cyprinidae ), roach, Rutilus rutilus European eel, Anguilla anguilla and herring, Clupea harengus, as well as a number of fish cell lines. Evidence is provided of the validation of these assays for specific species, a range of tissue types and cell lines as well as an example of the potential uses of these assays.