Development, application and validation of a Taqman real-time RT-PCR assay for the detection of infectious salmon anaemia virus (ISAV) in Atlantic salmon (Salmo salar).
Infectious salmon anaemia (ISA) is a disease of cultured Atlantic salmon (Salmo salar) which was successfully eradicated from Scotland following its emergence in 1998. The rapid deployment of sensitive diagnostic methods for the detection of ISA virus (ISAV) was fundamental to the swift eradication of ISA disease in Scotland and continues to be of crucial importance to surveillance of the aquaculture industry. This study reports the development, validation, application and interpretation of two independent, highly sensitive and specific semi-quantitative Taqman real-time RT-PCR (qRT-PCR) methods for the detection of ISAV. Such technology offers considerable advantages over conventional RT-PCR methods in current routine use for ISAV surveillance. These include an increased sensitivity, enhanced specificity, semi-quantification using endogenous controls, a lack of subjectivity in results interpretation, speed of processing and improved contamination control.
Snow
M
M
FRS Marine Laboratory, Aberdeen, UK. M.Snow@marlab.ac.uk
McKay
P
P
McBeath
A J A
AJ
Black
J
J
Doig
F
F
Kerr
R
R
Cunningham
C O
CO
Nylund
A
A
Devold
M
M
D000818:Animals; D005393:Fish Diseases / Q000175:diagnosis* / Q000453:epidemiology / Q000821:virology; D005880:Gills / Q000821:virology; D046829:Isavirus / Q000235:genetics* / Q000302:isolation & purification*; D007668:Kidney / Q000821:virology; D009976:Orthomyxoviridae Infections / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D020317:Salmo salar / Q000821:virology*; D012606:Scotland / Q000453:epidemiology; D012680:Sensitivity and Specificity