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首页> 外文期刊>The Analyst: The Analytical Journal of the Royal Society of Chemistry: A Monthly International Publication Dealing with All Branches of Analytical Chemistry >Detection of cystic fibrosis transmembrane conductance regulator Delta F508 gene mutation using a paper-based nucleic acid hybridization assay and a smartphone camera
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Detection of cystic fibrosis transmembrane conductance regulator Delta F508 gene mutation using a paper-based nucleic acid hybridization assay and a smartphone camera

机译：一种使用纸核酸杂交测定和智能手机相机检测囊性纤维化跨膜电导调节剂ΔF508基因突变

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Diagnostic technology that makes use of paper platforms in conjunction with the ubiquitous availability of digital cameras in cellular telephones and personal assistive devices offers opportunities for development of bioassays that are cost effective and widely distributed. Assays that operate effectively in aqueous solution require further development for implementation in paper substrates, overcoming issues associated with surface interactions on a matrix that offers a large surface-to-volume ratio and constraints on convective mixing. This report presents and compares two related methods for determination of oligonucleotides that serve as indicators of cystic fibrosis, differentiating between the normal wild-type sequence, and a mutant-type sequence that has a 3-base replacement. The transduction strategy operates by selective hybridization of oligonucleotide probes that are conjugated to fluorescent quantum dots, where hybridization of target sequences causes a molecular fluorophore to approach the quantum dot and become emissive through fluorescence resonance energy transfer Detection can rely on hybridization of a target that is labelled with Cy3 fluorophore, or in the presence of an unlabelled target when a sandwich assay format is implemented with a labelled reporter oligonucleotide. Selectivity to determine the presence of mismatched sequences involves appropriate selection of nucleotide sequences to set melt temperatures, in conjunction with control of stringency conditions using formamide as a chaotrope. It was determined that both direct and sandwich assays on paper substrates are able to distinguish between wild-type and mutant-type samples.
机译：利用纸张平台与蜂窝电话和个人辅助设备中的数码相机无处不在的可用性使用诊断技术为生物测定的发展提供了成本效益和广泛分布的机会。在水溶液中有效操作的测定需要进一步发展在纸质基质中的实施，克服与基质上的表面相互作用相关的问题，其为对流混合提供大的面对体积比和约束。该报告提供了两种相关方法，用于测定用作囊性纤维化指标的寡核苷酸，区分正常的野生型序列和具有3碱基替代的突变型序列。转导策略通过选择性杂交的寡核苷酸探针杂交，所述寡核苷酸探针与荧光量子点缀合，其中靶序列的杂交导致分子荧光团接近量子点并通过荧光共振能量转移检测变为发射，可以依赖于目标的杂交用Cy3荧光团标记，或者当用标记的报告器寡核苷酸实施夹层测定形式时，在未标记的靶标记中。 Selectivity to determine the presence of mismatched sequences involves appropriate selection of nucleotide sequences to set melt temperatures, in conjunction with control of stringency conditions using formamide as a chaotrope.确定纸质基材的直接和夹心测定能够区分野生型和突变型样品。

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