No abstract

A n o n r a d~o a c t~v e DNA probe assay was developed to detect and ~d e n t~f y infect~ous hernatopoiet~c necrosls virus (IHNV) uslng a dot blot format The probe a synthet~c DNA oligonucleot~de labeled enzymatlcally w~t h biotln h y b n d~z e d spec~f~cally w~t h nucleocaps~d mRNA extracted from Infected cells early In the vlrus repl~cation cycle A r a p~d g u a n~d l n~u m th~ocyanate based RNA extraction method uslng RNAzol B and rn~crocentrifuge tubes e f f~c~e n t l y pioduced h~g h q u a l~t y RNA from 3 commonly used f~s h cell llnes, CHSE-214, CHH-1, and EPC The probe reacted with 6 d~v e r s e ~solates of IHNV, but d~d not react \ n t h 2 related rhabdovlruses of fish viral hemorrhagic septlcemla vlrus and H~r a m e rhabdovlrus The b~o t~n y l a t e d probe was sensltlve d e t e c t~n g plcogram levels of target mRNA Detect~on and ~d e n t i f~c a t~o n of IHNV r e q u~r e d 2 d when cells wele lnoculated at n~u l t i p l i c~t~e s of infect~on (MOI) greater than 2 Flve days were necessary to detect and identify IHNV In cells lnoculated at a MO1 of 0 0002