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首页> 外文期刊>Fisheries and Aquatic Sciences >Development of a Denaturing High-Performance Liquid Chromatography (DHPLC) Assay to Detect Parasite Infection in Grass Shrimp Palaemonetes pugio
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Development of a Denaturing High-Performance Liquid Chromatography (DHPLC) Assay to Detect Parasite Infection in Grass Shrimp Palaemonetes pugio

机译：变性高效液相色谱（DHPLC）检测试剂盒的开发，用于检测草虾中的寄生虫感染Palaemonetes pugio

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摘要

In developing a useful tool to detect parasitic dynamics in an estuarine ecosystem, a denaturing high-performance liquid chromatography (DHPLC) assay was optimized by cloning plasmid DNA from the grass shrimp Palaemonetes pugio, and its two parasites,the trematode Microphallus turgidus and bopyrid isopod Probopyrus pandalicola. The optimal separation condition was an oven temperature of 57.9°C and 62-68% of buffer B gradient at a flow rate of 0.45 mL/min. A peptide nucleic acid blocking probe was designed to clamp the amplification of the host gene, which increased the amplification efficiency of genes with low copy numbers. Using the DHPLC assay with wild-type genomic, the assay could detect GC Gram positive bacteria and the bopyrid isopod (P. pandalicola). Therefore, the DHPLC assay is an effective tool for surveying parasitic dynamics in an estuarine ecosystem.
机译：在开发检测河口生态系统中寄生虫动态的有用工具时，通过从草虾对虾（Palaumonetes pugio）及其两个寄生虫（吸虫线虫Microphallus turgidus和bopyrid isoopod）中克隆质粒DNA，优化了变性高效液相色谱（DHPLC）测定法Probopyrus pandalicola。最佳分离条件为柱箱温度为57.9°C，流速为0.45 mL / min，缓冲液B梯度为62-8％。设计了一种肽核酸阻断探针来钳制宿主基因的扩增，从而提高了低拷贝数基因的扩增效率。使用具有野生型基因组的DHPLC测定法，该测定法可以检测GC革兰氏阳性细菌和bopyrid异足动物（P. pandalicola）。因此，DHPLC分析是一种用于调查河口生态系统中寄生动态的有效工具。

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