Rapid Detection of Infectious Pancreatic Necrosis Virus (IPNV) by the Enzyme-linked Immunosorbent Assay (ELISA)

An enzyme-linked immunosorbent assay (ELISA) has been developed for identification of the Sp strain of infectious pancreatic necrosis virus (IPNV). This assay employed 2 monoclonal antibodies directed against 2 non-overlapping epitopes of the minor structural viral protein.It was used to demonstrate the virus antigen in cell cultures and was able to detect 10 ng ml-' of purified virus or 104 TCIDSo ml-l in tissue culture fluid. The assay could be reduced to one step and b e performed within 90 min with good sensitivity. No antigenic variation, affecting the epitopes recognized by these monoclonal antibodies, was observed on 17 IPNV isolates of Sp serotype tested.

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Use of monoclonal antibodies for detection of infectious pancreatic necrosis virus by the enzyme-linked immunosorbent assay (ELISA)

Domínguez¹,

Hedrick²,

Sánchez-Vizcaíno³

1990

“…Virus identification. When CPE was observed in cell cultures inoculated with fish extracts, supernatant fluids were tested in enzyme-linked immunosorbent assays (ELISAs) for birnaviruses (Dixon & Hill 1983) Fish sampling. Fish were caught at 10 sampling (as they are ubiquitous in the marine environment) areas in UK coastal waters ( Fig.…”

Section: Sampling Areasmentioning

confidence: 99%

Isolation of viral haemorrhagic septicaemia virus from Atlantic herring Clupea harengus from the English Channel

Dixon¹,

Feist²,

Kehoe³

et al. 1997

Viral haemorrhaglc septicaemia vuus (VHSV) was Isolated from apparently healthy Atlantlc herrlng Clupea harengus from the Engllsh Channel The virus was lsolated in blueglll flbroblast (BF-2) cells but not eplthelioma papulosum cypnni (EPC) cells, however, the vlrus was passaged from BF-2 to EPC cells The identlty of the vlrus was confirmed by an enzyme-llnked ~rnmunosorbent assay and by a reverse transcnptase-polymerase chain reactlon (RT-PCR) The vlrus was classifled as a member of genogroup 111 of VHSV which compnses many European freshwater and m a n n e VHSV 150-lates Sequence compansons oi the RT-PCR products showed that the hernng isolate was closely related (99 1 % nucleotlde similarity) to a VHSV isolate from Atlantlc cod Gadus morhua both of those lsolates were avlrulent for ralnbow trout Oncorhynchus myklss by bath infection Circumstantial evidence IS presented whlch suggests that VHSV from hernng could have been the source of VHS dlsease In ralnbow trout in Denmark

“…Cell cultures showing CPE suggestive of rhabdoviruses or birnaviruses were tested in ELISAs for VHSV, infectious haematopoietic necrosis virus (IHNV) (Way & Dixon 1988, Way 1996 and birnavirus Serotype A2 (Dixon & Hill 1983) to ascertain the presence of those viruses. The latter ELISA was modified by changing the enzyme from alkaline phosphatase to horseradish peroxidase and then performed as described by Way & Dixon (1988) and Way (1996).…”

Section: Introductionmentioning

confidence: 99%

Four years of monitoring for viral haemorrhagic septicaemia virus in marine waters around the United Kingdom

Dixon

Avery²,

Chambers³

et al. 2003

Between 1995 and 1998, marine fish from around the coast of the UK were collected and samples analysed for viral haemorrhagic septicaemia virus (VHSV) using cell culture isolation methods. In 1997 and 1998 the samples were also analysed for VHSV by reverse transcription PCR (RT-PCR). A total of 1867 fish of 11 species were tested, but VHSV was isolated on only 1 occasion, from herring Clupea harengus, in 1996 . However, despite VHSV not being isolated in 1997 and 1998, in both years samples of herring from the west and south coasts of England produced positive signals in the RT-PCR, and in 1997 cod from the east coast of England also produced positive signals in the RT-PCR. These results are believed to be true indications of the presence of VHSV nucleic acid in the fish. In 1997, birnaviruses from Serogroup B1 were isolated from herring (a previously unrecorded host for the virus) and cod Gadus morhua, and a birnavirus from Serogroup A2 was also isolated from cod. In 1998, an aquareovirus was isolated from haddock Melanogrammus aeglefinus, a previously unrecorded host for the virus.