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首页> 外文期刊>Harmful Algae >Development of a qualitative PCR method for the Alexandrium spp. (Dinophyceae) detection in contaminated mussels (Mytilus galloprovincialis)
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Development of a qualitative PCR method for the Alexandrium spp. (Dinophyceae) detection in contaminated mussels (Mytilus galloprovincialis)

机译：亚历山大藻属定性PCR方法的开发。 （贻贝科）检测受污染的贻贝（Mytilus galloprovincialis）

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摘要

Paralytic shellfish poisoning (PSP) is a syndrome caused by the consumption of shellfish contaminated with neurotoxins produced by organisms of the marine dinoflagellate genus Alexandrium. A. minutum is the most widespread species responsible for PSP in the Western Mediterranean basin. The standard monitoring of shellfish farms for the presence of harmful algae and related toxins usually requires the microscopic examination of phytoplankton populations, bioassays and toxin determination by HPLC. These procedures are time-consuming and require remarkable experience, thus limiting the number of specimens that can be analyzed by a single laboratory unit. Molecular biology techniques may be helpful in the detection of target microorganisms in field samples. In this study, we developed a qualitative PCR assay for the rapid detection of all potentially toxic species belonging to the Alexandrium genus and specifically A. minutum, in contaminated mussels. Alexandrium genus-specific primers were designed to target the 5.8S rDNA region, while an A. minutum species-specific primer was designed to bind in the ITS 1 region. The assay was validated using several fixed seawater samples from the Mediterranean basin, which were analyzed using PCR along with standard microscopy procedures. The assay provided a rapid method for monitoring the presence of Alexandrium spp. in mussel tissues, as well as in seawater samples. The results showed that PCR is a valid, rapid alternative procedure for the detection of target phytoplankton species either in seawater or directly in mussels, where microalgae can accumulate.
机译：麻痹性贝类中毒（PSP）是由于食用被海洋鞭毛藻类亚历山大藻属的生物体所产生的神经毒素污染的贝类而引起的综合症。细叶农杆菌是造成西地中海盆地中PSP的最广泛物种。对贝类养殖场中有害藻类和相关毒素的存在进行标准监测通常需要对浮游植物种群进行显微镜检查，生物测定法和通过HPLC测定毒素。这些过程很耗时，并且需要出色的经验，因此限制了单个实验室单位可以分析的样本数量。分子生物学技术可能有助于检测野外样品中的目标微生物。在这项研究中，我们开发了一种定性PCR分析法，用于快速检测受污染的贻贝中所有属于亚历山大属的植物，尤其是小曲霉。设计亚历山大属属特异性引物以靶向5.8S rDNA区域，而拟南芥物种种特异性引物设计为结合在ITS 1区域。使用来自地中海盆地的几个固定海水样品验证了该测定，并使用PCR和标准显微镜程序对其进行了分析。该测定提供了监测亚历山大藻属的快速方法。在贻贝组织以及海水样本中。结果表明，PCR是检测海水或直接在贻贝中积聚微藻的浮游植物目标物种的有效，快速替代方法。

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