A NOVEL HIGH-THROUGHPUT SNP DETECTION APPROACH BASED MAGNETIC PARTICLES AND DUAL-COLOR FLUORESCENCE HYBRIDIZATION

摘要

Single nucleotide polymorphisms (SNPs) in human genes cause various genetic disorders and SNPs assay is expected to elucidate the genetics of individual differences in drug response and disease susceptibility. Consequently, an extremely efficient method for validation of co-dominant SNP locus for many patients with sensitivity, highthroughput, easy operation and high accuracy is required. This paper described a rapid, effective and high-throughput method for SNP genotyping using streptavidin-covered magnetic particles (SA-MPs) as targets carriers. In this approach) no conventional purification and concentration of PCR products were involved, and also no special modification of substrates like glass slides was performed. The genotyping results were obtained by scanning the dual-color probes denatured and printed on an unmodified glass slide, in which the high scattering of MPs was avoided and the genotyping took only a few minutes to carry out. Moreover, since all the procedures described in this study can be carried out in a 96 or 384-well PCR plate, this method can be developed to be an automatic genotyping system by using the automated workstation, arrayer and scanner. Because the method has a high sensitivity, a high signal intensity and a high match-to-mismatch signal ratio for SNP genotyping as reported in this paper, it will has a wide potential application in research and medical diagnosis.