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量化多种检测平台的水生入侵物种的环境 DNA 信号。

Quantifying environmental DNA signals for aquatic invasive species across multiple detection platforms.

机构信息

Department of Biology, Institute for Great Lakes Research, Central Michigan University , Mount Pleasant, Michigan 48859 United States.

出版信息

Environ Sci Technol. 2014 Nov 4;48(21):12800-6. doi: 10.1021/es5034052. Epub 2014 Oct 23.

Abstract

The use of molecular surveillance techniques has become popular among aquatic researchers and managers due to the improved sensitivity and efficiency compared to traditional sampling methods. Rapid expansion in the use of environmental DNA (eDNA), paired with the advancement of molecular technologies, has resulted in new detection platforms and techniques. In this study we present a comparison of three eDNA surveillance platforms: traditional polymerase chain reaction (PCR), quantitative PCR (qPCR), and digital droplet PCR (ddPCR) in which water samples were collected over a 24 h time period from mesocosm experiments containing a population gradient of invasive species densities. All platforms reliably detected the presence of DNA, even at low target organism densities within the first hour. The two quantitative platforms (qPCR and ddPCR) produced similar estimates of DNA concentrations. The analyses completed with ddPCR was faster from sample collection through analyses and cost approximately half the expenditure of qPCR. Although a new platform for eDNA surveillance of aquatic species, ddPCR was consistent with more commonly used qPCR and a cost-effective means of estimating DNA concentrations. Use of ddPCR by researchers and managers should be considered in future eDNA surveillance applications.

摘要

由于与传统采样方法相比具有更高的灵敏度和效率,分子监测技术在水生态研究人员和管理者中变得越来越流行。环境 DNA(eDNA)的使用迅速扩展,加上分子技术的进步,产生了新的检测平台和技术。在本研究中,我们比较了三种 eDNA 监测平台:传统聚合酶链反应(PCR)、定量 PCR(qPCR)和数字液滴 PCR(ddPCR),这些平台在包含入侵物种密度梯度的中观实验中,在 24 小时的时间内收集水样。所有平台都可靠地检测到了 DNA 的存在,即使在最初的一个小时内目标生物的密度很低也是如此。两种定量平台(qPCR 和 ddPCR)产生了相似的 DNA 浓度估计值。ddPCR 完成的分析从样本采集到分析的速度更快,花费约为 qPCR 的一半。ddPCR 虽然是一种新的水生物种 eDNA 监测平台,但与更常用的 qPCR 一致,是一种估算 DNA 浓度的经济有效方法。研究人员和管理者应该在未来的 eDNA 监测应用中考虑使用 ddPCR。

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