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首页> 外文期刊>Harmful Algae >A DNA hybridization assay to identify toxic dinoflagellates in coastal waters: detection of Karenia brevis in the Rookery Bay National Estuarine Research Reserve
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A DNA hybridization assay to identify toxic dinoflagellates in coastal waters: detection of Karenia brevis in the Rookery Bay National Estuarine Research Reserve

机译：DNA杂交测定法可识别沿海水域中的有毒鞭毛藻：在新库里湾国家河口研究保护区中检测到短毛克雷恩菌

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摘要

A DNA hybridization assay was developed in microtiter plate format to detect the presence of toxic dinoflagellates in coastal waters. Simultaneous detection of multiple species was demonstrated using Karenia brevis, Karenia mikimotoi, and Amphidinium carterae. Molecular probes were designed to detect both K. brevis and K. mikimotoi and to distinguish between these two closely related species. The assay was used to detect K. brevis in coastal waters collected from the Rookery Bay National Estuarine Research Reserve. Assay results were verified by species-specific PCR and sequence analysis. The presence/absence of K. brevis was consistent with microscopic observation. Assay sensitivity was sufficient to detect K. brevis in amounts defined by a regional monitoring program as "present" ( ≤ 1000 cells/L). The assay yielded quick colorimetric results, used a single hybridization temperature, and conserved the amount of genomic DNA utilized by employing one set of PCR primers. The microplate assay provides a useful tool to quickly screen large sample sets for multiple target organisms.
机译：以微量滴定板形式开发了一种DNA杂交测定法，以检测沿海水域中有毒的鞭毛藻的存在。同时证明了使用短的卡列氏菌（Karenia brevis），三叶卡列氏菌（Karenia mikimotoi）和角叉两栖菌（Amphidinium Carterae）可以同时检测多种物种。分子探针被设计用于检测短双歧杆菌和mikimotoi双歧杆菌，并区分这两个紧密相关的物种。该测定法用于检测从新鸟湾国家河口研究保护区收集的沿海水域中的短小K.brevis。通过物种特异性PCR和序列分析验证了测定结果。短杆菌K.的存在与否与显微镜观察一致。测定灵敏度足以检测区域监测程序定义为“存在”（≤1000个细胞/ L）的短小克氏杆菌。该测定产生快速的比色结果，使用单一杂交温度，并通过使用一组PCR引物节省了使用的基因组DNA量。微孔板测定法提供了一种有用的工具，可以快速筛查多种目标生物的大样本集。

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