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通过实时聚合酶链反应检测鲑肾杆菌诊断细菌性肾病

Diagnosis of bacterial kidney disease by detection of Renibacterium salmoninarum by real-time PCR.

作者信息

Jansson E, Lindberg L, Säker E, Aspán A

机构信息

Department of Wildlife, Fish and Environment, National Veterinary Institute, Uppsala, Sweden.

出版信息

J Fish Dis. 2008 Oct;31(10):755-63. doi: 10.1111/j.1365-2761.2008.00949.x. Epub 2008 Aug 4.

Abstract

Bacterial kidney disease (BKD), caused by Renibacterium salmoninarum (Rs), is a serious threat to salmon in aquaculture as well as to wild populations. We have developed a real-time polymerase chain reaction (PCR) for detection of Rs in kidney samples. The PCR is based on detection of unique parts of the 16S rRNA gene of Rs and DNA equivalent to 1-10 Rs genomes was detected per reaction. No cross-reactivity with other fish pathogenic or related bacteria could be demonstrated. Analysis of individual kidney samples collected from BKD classified populations identified 39.9% of the fish as positive by real-time PCR compared with 28.0% by polyclonal enzyme-linked immunosorbent assay (ELISA). The real-time PCR assay was found to be well suited for complementary use with ELISA for diagnosis of BKD, with the ability to detect clinical as well as covert Rs infections. The infection level determined by the polyclonal ELISA and by real-time PCR was significantly correlated.

摘要

由鲑肾杆菌(Rs)引起的细菌性肾病(BKD)对水产养殖中的鲑鱼以及野生种群构成严重威胁。我们开发了一种用于检测肾脏样本中Rs的实时聚合酶链反应(PCR)。该PCR基于对Rs 16S rRNA基因独特部分的检测,每个反应可检测到相当于1至10个Rs基因组的DNA。未证明与其他鱼类致病或相关细菌有交叉反应。对从BKD分类种群中采集的单个肾脏样本进行分析,通过实时PCR鉴定出39.9%的鱼为阳性,而通过多克隆酶联免疫吸附测定(ELISA)的阳性率为28.0%。发现实时PCR检测法非常适合与ELISA互补用于BKD的诊断,能够检测临床以及隐性Rs感染。通过多克隆ELISA和实时PCR确定的感染水平显著相关。

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