基于载体芯片的探针单核苷酸识别。

Single nucleotide recognition using a probes-on-carrier DNA chip.

机构信息

Area of Bioscience & Biotechnology, School of Materials Science, Japan Advanced Institute of Science & Technology (JAIST), 1-1 Asahidai, Nomi, Ishikawa 923-1292, Japan.

Kankyou Resilience, 79-7 Tokiwadai, Hodogaya, Yokohama, 240-0067, Japan.

出版信息

Biotechniques. 2019 Feb;66(2):73-78. doi: 10.2144/btn-2018-0088.

DOI:10.2144/btn-2018-0088

PMID:30744407

Abstract

Following the sequencing of the human genome, SNP analysis of individual patients has become essential for achieving the best drug response and ensuring optimal care. In this study, we developed a cost-effective probes-on-carrier DNA chip for the detection of SNPs. Our chips harbored three different probes against the TP53 gene, and were capable of detecting wild-type TP53 and SNPs such as rs121912651 and rs11540652. Four cell lines were used to validate the specificity of probe hybridization. Strong fluorescence intensity was observed in hybridized spots based on hybridization for perfect base pairing between complementary strands, whereas significantly lower fluorescence (p < 0.05) was observed in nonhybridized spots. These hybridization results indicated that the probes-on-carrier chip is suitable for SNP genotyping.

摘要

在人类基因组测序之后，对个体患者的 SNP 分析对于实现最佳药物反应和确保最佳护理至关重要。在这项研究中，我们开发了一种具有成本效益的载片探针 DNA 芯片，用于 SNP 的检测。我们的芯片上有针对 TP53 基因的三个不同探针，能够检测野生型 TP53 和 SNP ，如 rs121912651 和 rs11540652。使用四个细胞系来验证探针杂交的特异性。在互补链之间完美碱基配对的杂交基础上，观察到杂交斑点的荧光强度较强，而在非杂交斑点则观察到显著较低的荧光强度（p<0.05）。这些杂交结果表明，载片探针芯片适用于 SNP 基因分型。