Development of a biotinylated DNA probe for detection of infectious hematopoietic necrosis virus
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Abstract
A nonrad~oact~ve DNA probe assay was developed to detect and ~dent~fy infect~ous hernatopoiet~c necrosls virus (IHNV) uslng a dot blot format The probe a synthet~c DNA oligonucleot~de labeled enzymatlcally w~th biotln hybnd~zed spec~f~cally w~th nucleocaps~d mRNA extracted from Infected cells early In the vlrus repl~cation cycle A rap~d guan~dln~um th~ocyanate based RNA extraction method uslng RNAzol B and rn~crocentrifuge tubes eff~c~ently pioduced h~gh qual~ty RNA from 3 commonly used f~sh cell llnes, CHSE-214, CHH-1, and EPC The probe reacted with 6 d~verse ~solates of IHNV, but d~d not react \nth 2 related rhabdovlruses of fish viral hemorrhagic septlcemla vlrus and H~rame rhabdovlrus The b~ot~nylated probe was sensltlve detect~ng plcogram levels of target mRNA Detect~on and ~dentif~cat~on of IHNV requ~red 2d when cells wele lnoculated at n~ultiplic~t~es 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
| Publication type | Article |
|---|---|
| Publication Subtype | Journal Article |
| Title | Development of a biotinylated DNA probe for detection of infectious hematopoietic necrosis virus |
| Series title | Diseases of Aquatic Organisms |
| DOI | 10.3354/DAO011057 |
| Volume | 11 |
| Year Published | 1991 |
| Language | English |
| Publisher | Inter-Reseach |
| Contributing office(s) | Western Fisheries Research Center |
| Description | 9 p. |
| First page | 57 |
| Last page | 65 |
| Google Analytic Metrics | Metrics page |