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通过 qPCR 进行非致死性取样检测鲑鱼肾杆菌,用于诊断细菌性肾病。

Non-lethal sampling for the detection of Renibacterium salmoninarum by qPCR for diagnosis of bacterial kidney disease.

机构信息

National Veterinary Institute, Uppsala, Sweden.

出版信息

J Fish Dis. 2022 Jun;45(6):883-894. doi: 10.1111/jfd.13614. Epub 2022 Apr 1.

DOI:10.1111/jfd.13614
PMID:35363399
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9322471/
Abstract

Bacterial kidney disease (BKD), caused by Renibacterium salmoninarum (Rs), can be transmitted both horizontally and vertically and there is no available cure or prophylaxis. The control of BKD requires continuous surveillance, which is challenging in aquaculture as well as in programs for conservation and restoration of salmonid fish strains. BKD is a notifiable disease in Sweden and is monitored through the mandatory health control program using a polyclonal ELISA for detection of the Rs p57 protein in kidney. Fish must be killed for sampling, an obvious disadvantage especially regarding valuable broodfish. The present study shows that gill-/cloacal swabs collected in vivo for real-time PCR (qPCR ), allow a sensitive and specific detection of Rs. The sensitivity of qPCR was estimated to 97.8% (credible interval (ci) 93.8%-100%) compared to 98.3% (ci 92.7%-100%) and 48.8% (ci 38.8%-58.8%) of kidney samples for qPCR (qPCR ) and ELISA (ELISA ) respectively, by use of the Bayesian Latent Class Analysis (BLCA). Since the goal of the program is eradication of BKD the most sensitive test is preferrable. Using qPCR instead of ELISA will result in a lower false negative rate and can be useful for surveillance in aquaculture and in breeding programs with valuable fish. However, a higher false positive rate warrants confirmatory lethal testing before a previously Rs negative farm is subject to restrictions.

摘要

细菌性肾病(BKD)由鲑居尾孢菌(Renibacterium salmoninarum,Rs)引起,可水平和垂直传播,目前尚无有效的治疗或预防方法。BKD 的控制需要持续监测,这在水产养殖以及鲑鱼品种的保护和恢复计划中都具有挑战性。BKD 在瑞典是一种法定报告疾病,通过使用多克隆 ELISA 检测肾脏中的 Rs p57 蛋白的强制性健康控制计划进行监测。采样必须杀死鱼类,这对于有价值的亲鱼来说是一个明显的缺点。本研究表明,体内采集的鳃/泄殖腔拭子可用于实时 PCR(qPCR),从而可以敏感且特异地检测到 Rs。与 qPCR 相比,qPCR 的敏感性估计为 97.8%(可信区间(ci)93.8%-100%),而使用贝叶斯潜在类别分析(BLCA),qPCR 和 ELISA 的肾脏样本分别为 98.3%(ci 92.7%-100%)和 48.8%(ci 38.8%-58.8%)。由于该计划的目标是根除 BKD,因此最敏感的检测方法是首选。使用 qPCR 代替 ELISA 将降低假阴性率,并可用于水产养殖和有价值鱼类的繁殖计划中的监测。但是,较高的假阳性率需要在之前 Rs 阴性的农场受到限制之前进行确认性致死测试。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29f1/9322471/fafe3a5d9a0f/JFD-45-883-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29f1/9322471/854216682ff4/JFD-45-883-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29f1/9322471/70fe89296b49/JFD-45-883-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29f1/9322471/fafe3a5d9a0f/JFD-45-883-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29f1/9322471/854216682ff4/JFD-45-883-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29f1/9322471/70fe89296b49/JFD-45-883-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/29f1/9322471/fafe3a5d9a0f/JFD-45-883-g002.jpg

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本文引用的文献

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2
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Rev Sci Tech. 2021 Jun;40(1):19-28. doi: 10.20506/rst.40.1.3207.
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Evidence for the Use of Mucus Swabs to Detect in Brook Trout.
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