通过巢式逆转录聚合酶链反应检测海水中的传染性鲑鱼贫血病毒。

Detection of infectious salmon anemia virus in sea water by nested RT-PCR.

作者信息

Løvdal Trond, Enger Oivind

机构信息

Department of Microbiology, University of Bergen, Norway.

出版信息

Dis Aquat Organ. 2002 May 10;49(2):123-8. doi: 10.3354/dao049123.

DOI:10.3354/dao049123

PMID:12078980

Abstract

A method to detect low levels of infectious salmon anemia virus (ISAV) in environmental samples has been developed. The method is based on concentrating the viruses by tangential flow filtration and ultracentrifugation prior to amplification of the extracted viral RNA by nested reverse transcription polymerase chain reaction (RT-PCR). In the current investigation, seawater samples from salmon holdings were used for ISAV identification. ISAV was detected in seawater samples from a salmon holding site and from a vessel transporting salmon in 2 consecutive trials of the methodology. When known concentrations of ISAV were added to 21 of sea water, 5.5 viruses ml(-1) (corresponding to a tissue culture titer of TCID50 1.6 x 10(-2) ml(-1)) were detected using this method.

摘要

已开发出一种检测环境样本中低水平传染性鲑鱼贫血病毒（ISAV）的方法。该方法基于在通过巢式逆转录聚合酶链反应（RT-PCR）扩增提取的病毒RNA之前，通过切向流过滤和超速离心浓缩病毒。在当前研究中，来自鲑鱼养殖场的海水样本用于ISAV鉴定。在该方法的连续2次试验中，在来自一个鲑鱼养殖场所和一艘运输鲑鱼船只的海水样本中检测到了ISAV。当向21份海水中添加已知浓度的ISAV时，使用该方法检测到5.5个病毒/ml（相当于组织培养半数感染剂量TCID50为1.6×10⁻²/ml）。