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首页> 外文期刊>Biosensors & Bioelectronics: The International Journal for the Professional Involved with Research, Technology and Applications of Biosensers and Related Devices >Multipurpose high-throughput filtering microarrays (HiFi) for DNA and protein assays
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Multipurpose high-throughput filtering microarrays (HiFi) for DNA and protein assays

机译：用于DNA和蛋白质测定的多功能高通量过滤微阵列（HiFi）

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摘要

We are reporting here a low cost colorimetric device for high-throughput multiplexed blood group genotyping and allergy diagnosis, displayed as an automated 96-well microtiter plate format. A porous polymeric membrane sealed at the bottom of each well accounts for the sensor support. For each sensing unit, a 6×6 matrix of specific probes is spotted on the external surface of the membrane resulting in 5mm~2 microarrays. Thanks to the membrane porosity, reagents dispensed into the well can be eliminated through vacuum soaking. This unusual design drastically reduces the assay background signal.The system was first validated on robust models composed of either two complementary oligonucleotide sequences or one allergen/specific rabbit IgG pair. The quality of both oligonucleotide and protein immobilisation on the membrane substrate was then demonstrated together with the capacity to use the arrayed biomolecules as probes for the quantitative detection of specific targets (respectively complementary oligonucleotide and specific antibody).On the basis of these good results, two multiplex assays were developed for crude biological samples testing, focussing on two human in vitro diagnosis applications: a hybridisation assay for multiplex blood group genotyping and a multiparametric immunoassay for allergy diagnosis. In both cases, the transfer to crude biological samples testing was successful i.e. high signal to noise ratio of the stained membranes, reproducibility and good correlation with results obtained using routine testing procedures.
机译：我们在这里报告了一种低成本的比色仪，用于高通量多重血型基因分型和过敏诊断，显示为自动96孔微量滴定板格式。密封在每个孔底部的多孔聚合物膜构成了传感器支撑。对于每个传感单元，在膜的外表面上点一个6×6的特定探针矩阵，形成5mm〜2个微阵列。由于膜的多孔性，可以通过真空浸泡消除分配到孔中的试剂。这种不寻常的设计极大地降低了测定的背景信号。该系统首先在由两个互补寡核苷酸序列或一个变应原/特异性兔IgG对组成的稳健模型上进行了验证。然后证明了寡核苷酸和蛋白质固定在膜基质上的质量，以及使用阵列化的生物分子作为定量检测特定靶标（分别是互补的寡核苷酸和特异性抗体）的探针的能力。开发了两种用于粗生物样品测试的多重测定法，主要针对两种人类体外诊断应用：用于多重血型基因分型的杂交测定法和用于过敏诊断的多参数免疫测定法。在这两种情况下，均已成功转移到粗生物样品测试，即染色膜的信噪比高，再现性好，并且与使用常规测试程序获得的结果具有良好的相关性。

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