In the world wide, food poisoning accidents related to Vibrio spp. are on the rise, even numbers of food poisoning by other foodborne pathogens are decreasing. Therefore, the requirement of the rapid, sensitive and convenient detection method for V. parahaemolyticus has been grown. The objective of this study is to develop a colorimetric loop-mediated isothermal amplification (LAMP) assay using a molecular beacon (HRPzyme connected with complementary oligonucleotides at the 5' and 3' ends) for the rapid, sensitive, and convenient detection of V. parahaemolyticus. The colorimetric LAMP assay optimized at 58.8°C showed a detection limit of 1 × 100 CFU/mL and was confirmed to be specific to V. parahaemolyticus. The colorimetric LAMP assay can be finished within 1 h including DNA extraction step. The method was successfully applied to flatfish samples artificially inoculated with known amount of V. parahaemolyticus, and its cut-off value for the flatfish samples was 1 × 101 CFU/g. In addition, the colorimetric LAMP assay developed in the study was found to be able to correct false-positive results, which are known to be a disadvantage of conventional LAMP assays. Therefore, these results indicated that the colorimetric LAMP method is a useful tool for the rapid, sensitive and convenient detection of V. parahaemolyticus in fishes and can also be used as a point-of-care molecular diagnostic technique since it does not require any expensive equipment such as a thermocycler and detectors.

中文翻译：

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基于HRP模拟分子信标的比色环介导的等温扩增（LAMP）测定法，用于快速检测溶血弧菌。

在世界范围内，食物中毒事故与弧菌属有关。呈上升趋势，甚至其他食源性病原体导致食物中毒的人数也在减少。因此，对于快速，灵敏，方便的副溶血性弧菌检测方法的需求日益增长。这项研究的目的是开发一种比色环介导的等温扩增（LAMP）测定法，该测定法使用分子信标（HRPzyme在5'和3'末端与互补寡核苷酸相连）来快速，灵敏和方便地检测V.副溶血。优化于58.8°C的比色LAMP测定法的检出限为1×100 CFU / mL，并被证实对副溶血弧菌具有特异性。比色LAMP分析可在1 h内完成，包括DNA提取步骤。该方法已成功应用于人工接种已知量副溶血性弧菌的比目鱼样品，其对比目鱼样品的临界值为1×101 CFU / g。另外，研究中开发的比色LAMP测定法被发现能够纠正假阳性结果，这是传统LAMP测定法的缺点。因此，这些结果表明比色LAMP方法是快速，灵敏和方便地检测鱼类中副溶血性弧菌的有用工具，并且由于不需要任何昂贵的方法，因此也可以用作即时点分子诊断技术。诸如热循环仪和探测器之类的设备。发现该研究中开发的比色LAMP测定法能够纠正假阳性结果，这是传统LAMP测定法的缺点。因此，这些结果表明比色LAMP方法是快速，灵敏和方便地检测鱼类中副溶血性弧菌的有用工具，并且由于不需要任何昂贵的方法，因此也可以用作即时点分子诊断技术。诸如热循环仪和探测器之类的设备。发现该研究中开发的比色LAMP测定法能够纠正假阳性结果，这是传统LAMP测定法的缺点。因此，这些结果表明比色LAMP方法是快速，灵敏和方便地检测鱼类中副溶血性弧菌的有用工具，并且由于不需要任何昂贵的方法，因此也可以用作即时点分子诊断技术。诸如热循环仪和探测器之类的设备。