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

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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