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建立一种定量 PCR 检测方法,用于检测和计数潜在产雪卡毒素的藻类——Gambierdiscus lapillus(旋沟藻目,甲藻门)。

Development of a quantitative PCR assay for the detection and enumeration of a potentially ciguatoxin-producing dinoflagellate, Gambierdiscus lapillus (Gonyaulacales, Dinophyceae).

机构信息

Climate Change Cluster (C3), University of Technology Sydney, Ultimo, New South Wales, Australia.

ithree institute (i3), University of Technology Sydney, Ultimo, New South Wales, Australia.

出版信息

PLoS One. 2019 Nov 15;14(11):e0224664. doi: 10.1371/journal.pone.0224664. eCollection 2019.

Abstract

Ciguatera fish poisoning (CFP) is an illness contracted through the ingestion of seafood containing ciguatoxins. It is prevalent in tropical regions worldwide, including in Australia. Ciguatoxins are produced by some species of Gambierdiscus. Therefore, screening of Gambierdiscus species identification through quantitative PCR (qPCR), along with the determination of species toxicity, can be useful in monitoring potential ciguatera risk in these regions. In Australia, CFP is prevalent in tropical Queensland and increasingly in sub-tropical regions of Australia, but has a report rate of approximately 10%. Yet the identity, distribution and abundance of ciguatoxin producing Gambierdiscus spp. is largely unknown. In this study, we developed a rapid qPCR assay to quantify the presence and abundance of Gambierdiscus lapillus, a likely ciguatoxic species first described from Australia. We assessed the specificity and efficiency of the qPCR assay. The assay was tested on 25 environmental samples from the Heron Island reef in the southern Great Barrier Reef, a ciguatera endemic region, to determine the presence and patchiness of this species across samples from Chnoospora sp., Padina sp. and Sargassum sp. macroalgal hosts.

摘要

雪卡鱼中毒(CFP)是一种通过摄入含有雪卡毒素的海鲜而感染的疾病。它在全球热带地区都很常见,包括澳大利亚。雪卡毒素是由某些 Gambierdiscus 物种产生的。因此,通过定量 PCR(qPCR)筛选 Gambierdiscus 物种鉴定,以及确定物种毒性,可以在这些地区监测潜在的雪卡毒素风险。在澳大利亚,CFP 在热带昆士兰州流行,并在澳大利亚亚热带地区日益流行,但报告率约为 10%。然而,雪卡毒素产生 Gambierdiscus spp. 的身份、分布和丰度在很大程度上是未知的。在这项研究中,我们开发了一种快速 qPCR 测定法来定量检测 Gambierdiscus lapillus 的存在和丰度,这是一种最早在澳大利亚描述的可能具有雪卡毒性的物种。我们评估了 qPCR 测定法的特异性和效率。该测定法在大堡礁南部赫伦岛礁的 25 个环境样本中进行了测试,以确定该物种在 Chnoospora sp.、Padina sp. 和 Sargassum sp. 大型藻类宿主样本中的存在和分布情况。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cad5/6857910/165ad468ae99/pone.0224664.g001.jpg

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