Development of a Practical SNPs Typing Method by Using Elactro-ActiveArtificial DNAs

大多数人类DNA多态性是单核苷酸多态性（snp），这与各种常见疾病的风险和药物反应的差异密切相关。检测SNP的临床重要性激发了对许多能够区分SNP等位基因与正常等位基因的分析系统的研究。然而，现有的系统并不令人满意，特别是当应用于手持snp传感器时，这对最终用户很有用。最终的分析方案必须允许最终用户只需要直接将含有pcr扩增目标dna的样品添加到便携式设备中的低成本芯片上。我们已经报道了互补DNA的电化学检测，这是基于空穴运输与二茂铁-p-共轭DNA探针。这种方法允许无标记DNA与准“开/关”数字响应的snp无试剂的区分。然而，为了满足上述检测方案的标准，我们的方法应该被彻底保留。在电活性假核苷的合成成本、用探针在预先修饰的电极上对目标dna的原位杂交以及比普通PCR产生的更长的目标dna的适用性方面。在这里，我们研究了新的二茂铁-p-共轭DNA探针，它几乎满足了实现用户友好的SNP传感器的要求，并且与芯片上PCR技术高度兼容。我们已经制备了各种结构的DNA探针，并检测了它们对许多具有临床意义的过渡和翻转SNP靶序列的SNP识别能力，以实现上述实际应用的目标。因此，我们开发了一种新的电化学SNP分型系统，该系统由合成可行、多功能、廉价的电活性DNA探针组成。该系统有望满足临床医学专业最终用户所要求的分析方案标准。为了实现理想的snp传感器，我们正在进一步改进我们的方法。少

Most human DNA polymorphisms are single-nucleotide polymorphisms (SNPs), which critically relate to risks of various common diseases and differences in drug response. The clinical importance of detecting SNPs has inspired investigations into numerous assay systems that can discriminate SNP alleles from normal ones. However, the existing systems are not satisfactory, particularly when applied to a hand-held SNPs sensor, which is useful for end users. The ultimate assay protocol must allow end users to only need to directly add the sample that contains the PCR-amplified target DNAs onto a low-cost chip in a portable device.We have reported an electrochemical detection of complementary DNAs that is based on hole transport with ferrocene-p-conjugated DNA probes. This method allows a reagentless discrimination of label-free DNA from their SNPs with quasi "on/off" digital response. Nevertheless, to meet the criteria of the above-mentioned assay protocol, our method should thoroughly be remad … More e in terms of synthetic cost for the electroactive pseudonucleoside, in situ hybridization of target DNAs on premodified electrode with probes, and applicability to the longer target DNAs than are produced by common PCR. Here we investigated new ferrocene-p-conjugated DNA probes that almost satisfy the requirements for realizing a user-friendly SNP sensor that is highly compatible with PCR on-chip technologies.We have prepared new DNA probes of various structures and examined their SNP discrimination abilities for target sequences of many transition and transversion SNPs of clinical importance, with the goal of the above-mentioned practical use in mind. As a result, we have developed a new electrochemical SNP typing system composed of synthetically feasible and versatile, cheap electro-active DNA probes. This system is expected to meet the criteria of the assay protocol required by end users majoring in clinical medicines. Further improvements of our methodology are in progress for realizing an ideal SNPs sensor. Less