患“红肝病”鳗鲡疱疹病毒的分离和鉴定

Herpesvirus Isolation and Identification of the Cultured Eel with ‘Red Liver Disease'

摘要: 利用鳗鲡疱疹病毒(HVA)DNA聚合酶基因引物对患病双色鳗鲡和美洲鳗鲡进行PCR检测(PCR检测结果仅有双色鳗鲡),获得约394bp的目的条带,测序表明扩增出的条带为AHV的DNA聚合酶基因片段,与GeneBank公布的HQ992949、GU233800、FJ940765等6个鳗鲡疱疹病毒基因同源性为100%,与GU205167、AF363783鳗鲡疱疹病毒基因同源性为99%。将患病双色鳗鲡、美洲鳗鲡肝脏和肾脏组织的除菌滤液,接种EPC细胞,细胞7d出现CPE,双色鳗鲡株传代至15代,美洲鳗鲡株传代至5代。

Abstract: 394bp fragment was amplified and cloned from genomic DNA of the diseased A.Rostrataand A.bicolor bicolor.The nucleotide homology was 100% with DNA polymerase of 6eels herpesvirus(HQ992949,GU233800,FJ940765,EU349272,EU349271,AF333066)and 99% of 2eels herpesvirus(GU205167,AF363783).The suspension of gill,liver and kidney from diseased A.bicolor bicolor and diseased A.Rostrata which filtered through a filter membrane to get rid of bacteria was used to infected EPC cells,CPE appeared after 7d.Now we have cultivated 14 generations fromA.bicolor bicolor and 5generations fromA.Rostrata.