【摘要】 目的建立一种简便、省时、有效的N-ras突变检测的"凸背"引物荧光PCR新技术。方法对比"凸背"引物荧光PCR新技术和Sanger测序法:通过对混有不同比例的N-ras基因热突变G13D(GGT>GAT)质粒的样本进行对照检测来比较两者的灵敏度;对97例急性髓细胞白血病(AML)患者的外周血的DNA进行N-ras的G13D突变检测,统计两者的符合率。结果 "凸背"引物荧光PCR新技术能检测出混有5%突变质粒型的样本,其灵敏度达5%,高于Sanger测序法的10%~20%。在97例AML患者的外周血的DNA中,"凸背"引物荧光PCR新方法和Sanger测序法检测到N-ras基因G13D突变分别为19例和18例,符合率达94.7%。结论 "凸背"引物荧光PCR新技术比测序法更为快速、简单,有更高的灵敏度,容易实现。更多还原

【Abstract】 Objective To establish a simple,rapid and effective "convex dorsal" primer fluorescence PCR to detect the mutation of N-ras.Methods The N-ras mutation of G13D(GGT >GAT) was detected by "convex dorsal" primers fluorescent PCR method and Sanger sequencing method,respectively.The samples which were mixed with different proportions of the N-ras mutant plasmid(two methods) were detected,and their coincidence rates were calculated.97 acute myeloid leukemia(AML)peripheral blood samples were tested,and their coincidence rate was calculated.Results The samples which were mixed with 5%N-ras mutant plasmid can be detected by the "convex dorsal" primers fluorescent PCR method.Its sensitivity(5%) was higher than Sanger sequencing method’s.In 97 acute myeloid leukemia(AML) peripheral blood samples,19 case with N-ras mutations(G13D) were detected by "convex dorsal" primers fluorescent PCR method,and 18 case with N-ras mutations(G13D) were detected by Sanger sequencing method.The coincidence rate is 94.7%.Conclusion The "convex dorsal" primers fluorescent PCR method is more convenient,high sensitivity than sequencing method,and easily achieved.更多还原