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Molecular Techniques for the Detection of Organisms in Aquatic Environments, with Emphasis on Harmful Algal Bloom Species

机译:重点研究有害藻华物种在水生环境中检测生物的分子技术

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Molecular techniques to detect organisms in aquatic ecosystems are being gradually considered as an attractive alternative to standard laboratory methods. They offer faster and more accurate means of detecting and monitoring species, with respect to their traditional homologues based on culture and microscopic counting. Molecular techniques are particularly attractive when multiple species need to be detected and/or are in very low abundance. This paper reviews molecular techniques based on whole cells, such as microscope-based enumeration and Fluorescence In-Situ Hybridization (FISH) and molecular cell-free formats, such as sandwich hybridization assay (SHA), biosensors, microarrays, quantitative polymerase chain reaction (qPCR) and real time PCR (RT-PCR). Those that combine one or several laboratory functions into a single integrated system (lab-on-a-chip) and techniques that generate a much higher throughput data, such as next-generation systems (NGS), were also reviewed. We also included some other approaches that enhance the performance of molecular techniques. For instance, nano-bioengineered probes and platforms, pre-concentration and magnetic separation systems, and solid-phase hybridization offer highly pre-concentration capabilities. Isothermal amplification and hybridization chain reaction (HCR) improve hybridization and amplification techniques. Finally, we presented a study case of field remote sensing of harmful algal blooms (HABs), the only example of real time monitoring, and close the discussion with future directions and concluding remarks.
机译:用于检测水生生态系统中生物的分子技术已逐渐被认为是标准实验室方法的一种有吸引力的替代方法。与基于培养和显微计数的传统同源物相比,它们提供了更快,更准确的手段来检测和监测物种。当需要检测多种物种和/或丰度非常低时,分子技术特别有吸引力。本文概述了基于全细胞的分子技术,例如基于显微镜的枚举和荧光原位杂交(FISH)和无分子细胞的格式,例如夹心杂交测定(SHA),生物传感器,微阵列,定量聚合酶链反应( qPCR)和实时PCR(RT-PCR)。还将那些将一种或几种实验室功能组合到单个集成系统(片上实验室)中的技术以及产生更高吞吐量数据的技术(例如下一代系统(NGS))进行了审查。我们还包括其他一些增强分子技术性能的方法。例如,纳米生物工程探针和平台,预浓缩和磁分离系统以及固相杂交提供了高度的预浓缩能力。等温扩增和杂交链反应(HCR)改善了杂交和扩增技术。最后,我们提出了一个实时监测有害藻华(HABs)的案例,这是实时监测的唯一示例,并以未来的方向和结束语结束了讨论。

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