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Diseases of Aquatic Organisms

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DAO 28:229-233 (1997)  -  doi:10.3354/dao028229

Single-tube, non-interrupted reverse transcription PCR for detection of infectious pancreatic necrosis virus

Wang WS, Wi YL, Lee JS

An assay using a single-tube, non-interrupted reverse transcription-polymerase chain reaction (RT-PCR) was established for the detection of infectious pancreatic necrosis virus (IPNV). Two primer sets, Pr D and Pr F, which framed a region within the gene coding for IPNV VP2 protein were used to amplify specific fragments of the IPNV genome. Amplified products were detected in cultured chinook salmon embryo cells (CHSE-214) infected with different IPNV strain isolates including WB, Ja, Sp, Ab, VR299, 3372, MFK, and CV-HB-1, but not in uninfected CHSE-214 nor in cells infected with IHNV or infectious bursal disease virus (IBDV). The detection limit of this method was estimated with purified viral RNA from the WB strain of IPNV to be between 15 fg and 15 pg in ethidium bromide-stained gels.

RT-PCR · IPNV · Fish · Virus

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