Fish Pathology
Online ISSN : 1881-7335
Print ISSN : 0388-788X
ISSN-L : 0388-788X
Detection of Infections Pancreatic Necrosis Virus (IPNV) from Leukocytes of Carrier Rainbow Trout Oncorhynchus mykiss
Sylvia RodriguezMarta AlonsoSara I. Perez-Prieto
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2001 Volume 36 Issue 3 Pages 139-146

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Abstract

Blood and kidney leukocytes were purified from two-year-old rainbow trout carrying infectious pancreatic necrosis virus (IPNV) and the cells were processed to detect the virus by two methods : 1) indirect immunofluorescence stain followed by flow cytometry analysis, and 2) RT-PCR or nested-PCR amplification. These methods were compared with separate homogenization of visceral samples, followed by inoculation on cell cultures and seroneutralization, a procedure routinely employed by most disease diagnostic laboratories. IPNV was isolated by homogenization of samples in all seven specimens examined. The assay took between 7 and 28 days required for the appearance of cytopathic effects, which is usual in subclinical samples. From purified leukocytes, IPNV was detected by RT-PCR in five out of the seven specimens. The nested-PCR improved the sensitivity of the assay and gave positive results for all the fish. In addition, flow cytometry demonstrated the presence of IPNV in all the fish in 8-24 h. Examination of leukocytes is a practical way of detecting IPNV and much less time-consuming than the homogenization technique.

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© The Japanese Society of Fish Pathology
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