期刊
JOURNAL OF VIROLOGICAL METHODS
卷 211, 期 -, 页码 43-50出版社
ELSEVIER
DOI: 10.1016/j.jviromet.2014.10.007
关键词
Ostreid herpesvirus type 1; Crassostrea gigas; Pacific oyster; Viral infection; In situ hybridization; mRNA
资金
- EU projects Bivalive (FP7 KBBE) [266157]
- Moltraq (Era Net EMIDA)
An in situ hybridization protocol for detecting mRNAs of ostreid herpesvirus type 1 (OsHV-1) which infects Pacific oysters, Crassostrea gigas, was developed. Three RNA probes were synthesized by cloning three partial OsHV-1 genes into plasmids using three specific primer pairs, and performing a transcription in the presence of digoxigenin dUTP. The RNA probes were able to detect the virus mRNAs in paraffin sections of experimentally infected oysters 26 h post-injection. The in situ hybridization showed that the OsHV-1 mRNAs were mainly present in connective tissues in gills, mantle, adductor muscle, digestive gland and gonads. DNA detection by in situ hybridization using a DNA probe and viral DNA guantitation by real-time PCR were also performed and results were compared with those obtained using RNA probes. (C) 2014 Elsevier B.V. All rights reserved.
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