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用于数字PCR应用的微注射成型液滴生成系统。

Micro-injection Molded Droplet Generation System for Digital PCR Application.

作者信息

Jo Daae, Kim So Young, Kang Hyeon Woo, Pyo Sung Han, Jeong Nam Kyu, Bae Nam Ho, Lee Seok Jae, Kim Yong Tae, Lee Kyoung G

机构信息

National Nanofab Center, 291 Daehak-ro, Yuseong-gu, Daejeon, 34141 Republic of Korea.

Bio R&D Lab, BioTNS Co.Ltd, 19-12 Daehak-ro 76 beonan-gil, Daejeon, 34141 Republic of Korea.

出版信息

Biochip J. 2022;16(4):433-440. doi: 10.1007/s13206-022-00079-8. Epub 2022 Sep 6.

DOI:10.1007/s13206-022-00079-8
PMID:36091641
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9446600/
Abstract

Sensitive, effective, and quantitative analysis of infectious pathogens is an important task for the prevention of human health threats. Herein, we present an advanced approach to producing gene-encapsulated microdroplets for quantitative analysis using a micropatterned metal mold and injection molding technique with an automatically operated system. An injection molded microdroplet generation device was successfully fabricated with a minimum channel width of 30 μm and optimized to produce 100 μm diameter droplets. The optimized microchannel design and flow rate also enable the production of stable numbers of microdroplets (~ 16,000 droplets). To verify the applicability of our device and system to droplet-based digital PCR analysis, () O157:H7 was selected as a model bacterial pathogen, and the stx2 gene was amplified in the microdroplets. The generated microdroplets exhibit both chemical and mechanical stability, and our results are similar to those obtained by a commercially available method. Accordingly, the usefulness of the microdroplet generative device and system is confirmed as a simple, fast, and reliable tool for the quantitative molecular analysis of infectious diseases.

摘要

对传染性病原体进行灵敏、有效且定量的分析是预防人类健康威胁的一项重要任务。在此，我们展示了一种先进的方法，即使用微图案化金属模具和注塑技术以及自动操作系统来制备用于定量分析的基因封装微滴。成功制造出了一种注塑微滴生成装置，其最小通道宽度为30μm，并经过优化以产生直径为100μm的微滴。优化后的微通道设计和流速还能够产生稳定数量的微滴（约16,000个微滴）。为了验证我们的装置和系统在基于微滴的数字PCR分析中的适用性，选择了大肠杆菌O157:H7作为模型细菌病原体，并在微滴中扩增stx2基因。所生成的微滴具有化学和机械稳定性，我们的结果与通过市售方法获得的结果相似。因此，微滴生成装置和系统作为一种用于传染病定量分子分析的简单、快速且可靠的工具的实用性得到了证实。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/ad5822963c0d/13206_2022_79_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/9492485115a9/13206_2022_79_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/42e92d55206b/13206_2022_79_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/04a466e806f3/13206_2022_79_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/daec0bccba7d/13206_2022_79_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/ad5822963c0d/13206_2022_79_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/9492485115a9/13206_2022_79_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/42e92d55206b/13206_2022_79_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/04a466e806f3/13206_2022_79_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/daec0bccba7d/13206_2022_79_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f97c/9446600/ad5822963c0d/13206_2022_79_Fig5_HTML.jpg

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