目的 BRAF V600E基因痕量突变的筛查可以避免肿瘤患者无效治疗的情况出现.方法 用内部竞争性扩增片段提高野生抑制型(WTB)探针对野生型BRAF V600E基因的抑制,提高发生痕量突变的BRAF V600E基因型的检出率.结果 当模板DNA浓度在50～200 ng/μL时,构建的痕量基因突变实时荧光定量检测方法能够完全屏蔽BRA F V600E野生型基因扩增.该检测方法的灵敏度可以达到0.1％,符合基因痕量突变检测技术灵敏度的要求.在50例疑似结直肠癌患者的结直肠镜活检组织中,构建的该方法检测到BRA F V600E基因痕量突变的标本为8例(16.0％),具备较高的检出率.结论 构建的基因痕量突变的检测方法能够对临床标本中BRA F V600E基因痕量突变做快速、简便、低成本的定量分析.%Objective To study the screening of trace amount mutation of BRAF V 600E gene for avoiding the appearance of ineffective treatment in cancer patients .Methods The internal competitive amplification fragments were used to improve the inhibition of wild-type blocking (WTB) probe on wile-type BRAF V600E gene to increase the detection efficiency of BRAF V600E genotype of trace amount mutation occurrence .Re-sults When the template DNA concentration was 50 -200 ng/μL ,the constructed trace amount gene muta-tion real time fluorescence quantitative detection method could completely block the amplification of the wild-type BRAF V600E gene .The sensitivity of this assay reached as high as 0 .1％ ,which was in line with the sen-sitivity requirement for the gene trace amount mutation detection technique .In the colorectal biopsy tissues from 50 cases of suspected colorectal cancer ,8 cases (16 .0％ ) of BRAF V600E gene trace amount mutation were detected by using this constructed method ,which had higher detection rate .Conclusion The constructed gene trace amount mutation detection method can make the rapid ,simple and low cost quantitative analysis for BRAF V600E gene trace amount mutation in clinical samples .
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