等位基因特异性-PCR联合限制性内切酶消化法检测JAK2V617F突变  

Detection of JAK2V617F Mutation by Allele specific-PCR and Restriction Enzyme Digestion

作　　者：申徐良[1] 陈方平[1] 魏武[2] 张梅香[2] 史文芝[2] 秦小琪[2] 徐洪亮[2] 

机构地区：[1]中南大学湘雅医院,410008 [2]长治医学院附属和平医院血液科

出　　处：《长治医学院学报》2008年第1期6-8,共3页Journal of Changzhi Medical College

基　　金：山西省自然科学基金资助项目(20011072);山西省教育厅基金项目(2024)

摘　　要：目的:建立敏感特异的JAK2V617F点突变的临床检测方法。方法:基因组DNA从HEL细胞或正常志愿者外周血中提取。采用等位基因特异性-PCR(Allele specific-PCR,AS-PCR)和限制性内切酶消化的方法检测基因组中JAK2V617F突变。结果:本AS-PCR法可对JAK2基因扩增出364bp的内参照条带,当存在JAK2V617F突变时,又可以扩增出203bp的突变条带。只有野生型内参照条带364bp可被BsaXⅠ消化切割。结论:AS-PCR法和限制性内切酶法是检测JAK2V617F突变的敏感特异的检测方法,可以成功应用于临床检测。Objective：To establish a sensitive and specific test for JAK2V617F point mutation. Methods： Genomic DNA was isolated from HEL cells or Peripheral - blood cells from normal volunteer. Allele - specific polymerase chain reactions （AS - PCR） and restriction enzyme digestion were performed to detect the mutation in genomic DNA. Results：A control band（364 bp） can be obtained by AS- PCR, and a muted band （203 bp） can be obtained only when JAK2V617F was presented. Only wildtype control band （364 bp） can be digested by restriction enzyme BsaX Ⅰ . Conclusion：AS - PCR and restriction enzyme digestion were sensitive and specific tests for JAK2V617F point mutation, and can be successfully used for clinical test.

关 键 词：等位基因特异性-PCR JAK2V617F突变 限制性内切酶BsaXⅠ 

分 类 号：R394.33[医药卫生—医学遗传学]