定量聚合酶链反应和夹心杂交法监测赤潮杂藻的比较研究。

IF 2.7 3区 环境科学与生态学 Q3 ENVIRONMENTAL SCIENCES
Dianne I Greenfield, Kathryn J Coyne
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引用次数: 0

摘要

监测水生环境中的有害藻华(HABs)通常是通过光学显微镜来辅助的,尽管基于分子的方法可以加快物种检测,细胞量化,从而为管理反应提供早期预警。定量聚合酶链反应(qPCR)和夹心杂交测定(SHA)两种方法越来越多地用于赤潮监测,但它们在方案、遗传靶点、设备/供应和其他考虑方面存在差异。这对最终用户在选择工具整合到有害藻华监测程序时提出了挑战。作为回应,我们进行了一项为期多年的qPCR和SHA相对于显微镜监测赤石刺虫(Heterosigma akashiwo)的并行比较研究,赤石刺虫是一种在世界范围内造成鱼类死亡和水质受损的物种。本文总结了一套广泛的并排,实验室和现场测试的结果,通过qPCR和SHA来定量赤石芽孢杆菌细胞。实验范围、检出限、对保存样品的适用性以及培养赤斑潜蝇的生理条件(一天中的时间、生长期、营养水平)表明,qPCR和SHA的一致性普遍较强,尽管qPCR具有更大的动态范围(不需要均质液稀释),而SHA的检出限较低。在实验室细胞生长期间以及在田间开花发育期间,这两种测定都产生了极好的显微镜一致性。然而,在稳定下降生长和低硝酸盐条件下,qPCR和SHA细胞丰度数据比显微镜下少,表明衰老和营养胁迫过程中细胞核酸减少。实际上,qPCR和SHA都是负担得起的,但qPCR解决方案通常比SHA更商业化。研究结果对于考虑适合其有害藻华监测需要的方法选择的管理人员将是有价值的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Comparison of Two Molecular Methods, Quantitative Polymerase Chain Reaction and Sandwich Hybridization Assay, for Monitoring the Harmful Algal Bloom Species, Heterosigma akashiwo.

Monitoring for harmful algal blooms (HABs) in aquatic environments is commonly aided by light microscopy, though molecular-based approaches can expedite species detection, cell quantification, and therefore early warnings for management responses. Two methods, quantitative polymerase chain reaction (qPCR) and sandwich hybridization assay (SHA), are increasingly used for HAB monitoring, but they differ in terms of protocols, genetic targets, equipment/supplies, and other considerations. This presents a challenge to end-users when selecting tool(s) to integrate within HAB surveillance programs. In response, we conducted a multi-year, side-by-side comparison study between qPCR and SHA relative to microscopy for monitoring the raphidophyte Heterosigma akashiwo, a species responsible for fish kills and impaired water quality worldwide. This paper summarizes key findings from a broad suite of side-by-side, laboratory and field tests of H. akashiwo cell quantification by qPCR and SHA. Assay ranges, detection limits, applicability to preserved samples, and physiological conditions (time of day, growth phase, nutrient levels) of cultured H. akashiwo revealed generally strong qPCR-SHA agreement, though qPCR had a wider dynamic range (without homogenate dilution) while SHA displayed a lower detection limit. Both assays yielded excellent agreement with microscopy during cell growth in the laboratory as well as during bloom development in the field. However, qPCR and SHA cell abundance data were less than microscopy during stationary-decline growth and under low nitrate, indicating reduced cellular nucleic acid during senescence and nutrient stress. Pragmatically, both qPCR and SHA are affordable, but qPCR solutions are typically more available commercially than SHA. Study results will be valuable to managers considering methodological options that suit their HAB monitoring needs.

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来源期刊
Environmental Management
Environmental Management 环境科学-环境科学
CiteScore
6.20
自引率
2.90%
发文量
178
审稿时长
12 months
期刊介绍: Environmental Management offers research and opinions on use and conservation of natural resources, protection of habitats and control of hazards, spanning the field of environmental management without regard to traditional disciplinary boundaries. The journal aims to improve communication, making ideas and results from any field available to practitioners from other backgrounds. Contributions are drawn from biology, botany, chemistry, climatology, ecology, ecological economics, environmental engineering, fisheries, environmental law, forest sciences, geosciences, information science, public affairs, public health, toxicology, zoology and more. As the principal user of nature, humanity is responsible for ensuring that its environmental impacts are benign rather than catastrophic. Environmental Management presents the work of academic researchers and professionals outside universities, including those in business, government, research establishments, and public interest groups, presenting a wide spectrum of viewpoints and approaches.
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