A reliable RT-PCR-ELISA method for the detection of infectious pancreatic necrosis virus (IPNV) in farmed rainbow trout.
A new method, termed RT-PCR-ELISA, was evaluated for ease of use, reliability and sensitivity when detecting infectious pancreatic necrosis virus (IPNV) present in trout kidney tissue. The method had comparable sensitivity to existing PCR assays and could successfully detect 1.5 x 10(4) pfu IPNV in artificially contaminated trout kidney samples. The technique was easily established in a new laboratory and required no specialised equipment. The method had a high sample throughput capable of screening 96 samples per run, making the technique extremely time efficient. The RT-PCR-ELISA is a safe, quick, reliable technique, which has the potential for use as a standard virus detection method.
Milne
S A
SA
Department of Molecular and Cell Biology, School of Medical Sciences, University of Aberdeen, I.M.S Building, Foresterhill, Aberdeen AB25 2ZD, Scotland. s.a.milne@abdn.ac.uk
Gallacher
S
S
Cash
P
P
Porter
A J R
AJ
D000818:Animals; D018175:Birnaviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005393:Fish Diseases / Q000175:diagnosis* / Q000821:virology; D018061:Infectious pancreatic necrosis virus / Q000302:isolation & purification*; D007668:Kidney / Q000821:virology; D017686:Oncorhynchus mykiss / Q000821:virology*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity