ONE-STEP GENOTYPING BY ALLELE-SPECIFIC AMPLIFICATION FROM WHOLE BLOOD

摘要

Single nucleotide polymorphism (SNP) plays an important role in drug response and disease susceptibility. In this study, we developed a onestep genotyping method, in which only a step of allele-specific amplification from whole blood is needed. For each SNP, two allele-specific amplifications were conducted in a pair of tubes each containing 0. 5 fig/mL ethidium bromide. After then the allele-specific amplicons were directly detected by fluorescence imaging. Since each tube corresponds to a type of allele, we can easily determine the type of SNP by comparing the fluorescence intensities of the two PCR tubes. We successfully established allele-specific amplifications from whole blood for determining five polymorphisms in 141 health controls. The genotyping results are consistent with those obtained by sequencing method. This method is accurate, time-saving, cost-effective, and easy-todo, especially suitable for clinical prediction of disease susceptibility.