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Since isolated from rainbow trout in 1983 in Taiwan, the IPNV
(Infectious Pancreatic
Necrosis Virus) was identified in many kind of marine and fresh water fish
continuously. In addition to horizontal transmission between fish, the vertical
transmission via carrier and eggs causes a large number of
deaths of fingerlings.
Because the source of rainbow trout eggs completely depends on
importation, this
experiment is designed to build a rapid diagnostic protocol of
IPNV to prevent spread
of virus via carrier and eggs. There are two approach in this
experiment: (1)RT-PCR:
Utilize proteinase K to digest the virus packaged in the shell
membrane to explore the
viral nucleic acid, then detected using RT-PCR. (2) In situ
hybridization: The probe
was labelled with Digoxigenin, then applied in ' in situ
hybridization'. Applied the
probe to detect the early stage of traditional viral isolated
cell line cultivation, and the
sensitivity is 101-2 TCID50/ml. In the other hand, detected the
sections of organs of the
inoculated rainbow trout and eel fingerlings, and the positive
infected cells in the liver,
kidney, gut and skin were detected , and prove this technique
can be applied for
clinical diagnosis.
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