Filtration extraction method using a microfluidic channel for measuring environmental DNA
Filtration extraction method using a microfluidic channel for measuring environmental DNA
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使用微流体通道的过滤提取方法测量环境DNA
DOI：
10.1111/1755-0998.13657
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发表时间：
2022
影响因子：
7.7
通讯作者：
Doi Hideyuki
中科院分区：
文献类型：
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作者：
Fukuzawa Takashi;Kameda Yuichi;Nagata Hisao;Nishizawa Naofumi;Doi Hideyuki
广泛应用于生物监测的环境DNA（eDNA）方法仅限于实验室分析和处理。在本研究中，我们开发了一种使用微流体通道 Biryu 芯片 (BC) 的过滤/提取组件，以及一种过滤/提取方法（BC 方法），以最大限度地减少 DNA 提取和随后的 PCR 扩增所需的样品体积。我们使用水族馆和河水样本测试了 BC 方法的性能，并将其与 Sterivex 过滤/提取方法的性能进行了比较。我们观察到，使用BC法，用Sterivex法过滤体积的1/20-1/40即可获得相同浓度的提取DNA，这表明BC法可广泛用于eDNA测量。此外，我们使用移动 PCR 设备在 30 分钟内对 eDNA 进行了现场测量，证明使用 BC 方法可以轻松快速地进行过滤和提取。使用BC方法获得的PCR结果与使用Sterivex方法获得的结果相似。 BC方法需要的步骤较少；因此，可以降低DNA污染的风险。与移动 PCR 结合使用时，BC 方法可在采集水样后 30 分钟内轻松检测 eDNA，甚至可以在现场检测。
The environmental DNA (eDNA) method, which is widely applied in biomonitoring, is limited to laboratory analysis and processing. In this study, we developed a filtration/extraction component using a microfluidic channel, the Biryu‐Chip (BC), and a filtration/extraction method, the BC method, to minimize the volume of the sample necessary for DNA extraction and subsequent PCR amplification. We tested the performance of the BC method and compared it with that of the Sterivex filtration/extraction method using aquarium and river water samples. We observed that using the BC method, the same concentration of extracted DNA was obtained with 1/20–1/40 of the filtration volume of the Sterivex method, suggesting that the BC method can be widely used for eDNA measurement. In addition, we performed on‐site measurements of eDNA within 30 min using a mobile PCR device, demonstrating that filtration and extraction can be performed easily and quickly using the BC method. The PCR results obtained using the BC method were similar to those obtained using the Sterivex method. The BC method requires fewer steps; therefore, the risk of DNA contamination can be reduced. When combined with mobile PCR, the BC method can be applied to easily detect eDNA within 30 min from the collection of water sample, even on‐site.