Detection of Infections Pancreatic Necrosis Virus(IPNV) from Leukocytes of Carrier Rainbow Trout Oncorhynchus mykiss.

Rodrı́guez, Sergio; Alonso, M. Carmen; Pérez-Prieto, Sara I.

doi:10.3147/jsfp.36.139

Cited by 26 publications

(19 citation statements)

References 21 publications

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“…Both procedures are time‐consuming and rely on the use of cell cultures, but differences in sensitivity have been reported. Some authors have reported the detection of IPNV in rainbow trout leucocytes by flow cytometry (Rodriguez et al. 2001), but unfortunately this technology is not affordable for many laboratories.…”

Section: Discussionmentioning

confidence: 99%

Isolation in cell culture and detection by PCR‐based technology of IPNV‐like virus from leucocytes of carrier turbot, Scophthalmus maximus (L.)

Cutrín

López-Vázquez

Olveira

et al. 2005

Journal of Fish Diseases

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A non-destructive procedure was utilized to determine the infectious pancreatic necrosis virus (IPNV) status of an apparently healthy turbot broodstock. Blood samples were used to detect IPNV by reverse transcriptase-polymerase chain reaction (RT-PCR), Southern blot hybridization and nested PCR. In addition, viral isolation from turbot leucocytes was performed. Around 22% of the fish were IPNV positive by RT-PCR, and this increased to close to 60% when nested PCR was performed. The present report supports the use of blood samples for the detection of IPNV-like viruses in brood fish. In addition, we demonstrate that it is possible to isolate the virus from the blood of carrier fish, as a non-lethal detection method, although it is much less sensitive than RT-PCR and nested PCR as a IPNV-like strain was isolated from only five of the 15 blood sample pools assayed. The viral isolate was identified as type Dry Mills (genogroup I) by means of restriction fragment length polymorphisms and DNA sequencing.

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Section: Discussionmentioning

confidence: 99%

Isolation in cell culture and detection by PCR‐based technology of IPNV‐like virus from leucocytes of carrier turbot, Scophthalmus maximus (L.)

Cutrín

López-Vázquez

Olveira

et al. 2005

Journal of Fish Diseases

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“…The kidney seems to be an important organ for the uptake and expression of DNA vaccines since pDNA uptake and transgene expression have both been demonstrated in kidney macrophages from several fish species [27,31,32]. We previously demonstrated VP2 gene expression in the kidneys of brown trout 30 days post-vaccination when studying the innate immune response induced by the pcDNA-VP2 vaccine administered by injection [1]. Here we obtained similar results after oral delivery of the same plasmid, indicating that alginate microspheres are indeed good carriers for oral delivery.…”

Section: Vaccine Treatmentmentioning

confidence: 98%

Immunogenic and protective effects of an oral DNA vaccine against infectious pancreatic necrosis virus in fish

Heras

Saint-Jean

Pérez-Prieto

2010

Fish & Shellfish Immunology

132

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“…IPNV VP2 protein expressed in BF-2 cells was detected with an indirect immunofluorescence assay (IFA), and the fluorescent cells were both quantified by flow cytometry [36] and observed by UV microscopy. Briefly, after incubation of transfected and non-transfected BF-2 cells at -20 C for 3 days, cell monolayers were disrupted with trypsin (0.25%), fixed with 3.7% formaldehyde in PBS for 15 min and permeabilized with 0.1% Triton X-100 in PBS for 1 min.…”

Section: Immunofluorescence Assaymentioning

confidence: 99%

In vitro and in vivo immune responses induced by a DNA vaccine encoding the VP2 gene of the infectious pancreatic necrosis virus

Heras

Prieto

Saint-Jean

2009

Fish & Shellfish Immunology

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The work presented here describes the construction of a plasmid encoding the VP2 gene of the infectious pancreatic necrosis virus (IPNV), its expression in BF-2 cells and an evaluation of its activity in brown trout (Salmo trutta L) soon after injection. Preliminary experiments to evaluate the potential of the plasmid to induce neutralizing antibodies were also performed. We established a BF-2 cell line that expresses VP2 constitutively and we examined the infection of these VP2-transfected BF-2 cells with homologous and heterologous viruses. The expression kinetics of IFN, and of the IFN-induced genes Mx and ISG15, was also evaluated in brown trout over a 15 day interval, and quantified by real-time or semi-quantitative PCR. Type I IFN and Mx are markers of the non-specific innate immune response to viruses and they are involved in antiviral defences. Our results demonstrate that expression of the IPNV VP2 protein in BF-2 cells induces an antiviral state against IPNV and against the infectious haematopoietic necrosis virus (IHNV). In BF-2 infected cells, VP2 inhibited both the IPNV and IHNV-induced cytopathic effect to some extent, as well as the virus yield. In vivo, VP2 was expressed in haematopoietic tissues such as the head kidney of 7 month-old trout. In addition, it induced early immune responses and specific immunity 30 days after injection. IFN mRNA expression increased sharply on the 1st and 15th day post-injection and expression of other IFN-induced genes as Mx and ISG15 was also detected soon after vaccination of brown trout. Moreover, specific antibodies were detected 30 days after vaccination. These results suggest that the VP2 gene is a good candidate for the design of IPNV-DNA vaccines and to investigate the use of cytokines as co-stimulatory molecules.

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Detection of Infections Pancreatic Necrosis Virus(IPNV) from Leukocytes of Carrier Rainbow Trout Oncorhynchus mykiss.

Cited by 26 publications

References 21 publications

Isolation in cell culture and detection by PCR‐based technology of IPNV‐like virus from leucocytes of carrier turbot, Scophthalmus maximus (L.)

Isolation in cell culture and detection by PCR‐based technology of IPNV‐like virus from leucocytes of carrier turbot, Scophthalmus maximus (L.)

Immunogenic and protective effects of an oral DNA vaccine against infectious pancreatic necrosis virus in fish

In vitro and in vivo immune responses induced by a DNA vaccine encoding the VP2 gene of the infectious pancreatic necrosis virus

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