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通过紫外光照射将 DNA 探针直接固定在未经修饰的塑料上,并将其集成在微流控装置中,用于快速生物分析。

Direct immobilization of DNA probes on non-modified plastics by UV irradiation and integration in microfluidic devices for rapid bioassay.

机构信息

DTU Nanotech, Department of Micro- and Nanotechnology, Technical University of Denmark, Ørsteds Plads, Kgs. Lyngby, Denmark.

出版信息

Anal Bioanal Chem. 2012 Jan;402(2):741-8. doi: 10.1007/s00216-011-5459-4. Epub 2011 Oct 26.

DOI:10.1007/s00216-011-5459-4
PMID:22028019
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3249165/
Abstract

DNA microarrays have become one of the most powerful tools in the field of genomics and medical diagnosis. Recently, there has been increased interest in combining microfluidics with microarrays since this approach offers advantages in terms of portability, reduced analysis time, low consumption of reagents, and increased system integration. Polymers are widely used for microfluidic systems, but fabrication of microarrays on such materials often requires complicated chemical surface modifications, which hinders the integration of microarrays into microfluidic systems. In this paper, we demonstrate that simple UV irradiation can be used to directly immobilize poly(T)poly(C)-tagged DNA oligonucleotide probes on many different types of plastics without any surface modification. On average, five- and fourfold improvement in immobilization and hybridization efficiency have been achieved compared to surface-modified slides with aminated DNA probes. Moreover, the TC tag only costs 30% of the commonly used amino group modifications. Using this microarray fabrication technique, a portable cyclic olefin copolymer biochip containing eight individually addressable microfluidic channels was developed and used for rapid and parallel identification of Avian Influenza Virus by DNA hybridization. The one-step, cost-effective DNA-linking method on non-modified polymers significantly simplifies microarray fabrication procedures and permits great flexibility to plastic material selection, thus making it convenient to integrate microarrays into plastic microfluidic systems.

摘要

DNA 微阵列已成为基因组学和医学诊断领域最强大的工具之一。最近,人们对将微流控技术与微阵列相结合越来越感兴趣,因为这种方法在便携性、分析时间缩短、试剂消耗减少和系统集成度提高等方面具有优势。聚合物广泛用于微流控系统,但在这些材料上制造微阵列通常需要复杂的化学表面修饰,这阻碍了微阵列与微流控系统的集成。在本文中,我们证明简单的紫外辐射可以直接将聚(T)聚(C)标记的 DNA 寡核苷酸探针固定在许多不同类型的塑料上,而无需任何表面修饰。与带有氨基化 DNA 探针的表面修饰载玻片相比,固定化和杂交效率平均提高了五倍和四倍。此外,TC 标签的成本仅为常用氨基修饰的 30%。使用这种微阵列制造技术,开发了一种包含八个可单独寻址微流道的可携式环烯烃共聚物生物芯片,并用于通过 DNA 杂交快速平行鉴定禽流感病毒。在未修饰的聚合物上进行的一步、具有成本效益的 DNA 连接方法显著简化了微阵列制造过程,并允许对塑料材料选择具有很大的灵活性,从而便于将微阵列集成到塑料微流控系统中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/6d9d9661ceeb/216_2011_5459_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/f38473b34370/216_2011_5459_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/9c39b5f25486/216_2011_5459_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/e4d72873b584/216_2011_5459_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/01bb54547743/216_2011_5459_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/6d9d9661ceeb/216_2011_5459_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/f38473b34370/216_2011_5459_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/9c39b5f25486/216_2011_5459_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/e4d72873b584/216_2011_5459_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/01bb54547743/216_2011_5459_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f76b/3249165/6d9d9661ceeb/216_2011_5459_Fig5_HTML.jpg

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