Crispr-Cas12a在成人弥漫性胶质瘤异柠檬酸脱氢酶2基因突变检测中的应用

Application of Crispr-Cas12a in the detection of isocitrate dehydrogenase 2 gene mutation in adult diffuse gliomas

目的探讨Crispr-Cas12a方法对成人弥漫性胶质瘤异柠檬酸脱氢酶2(IDH2)基因R172K单核苷酸多态性的检测效能。方法选择2019年1月至2022年9月手术切除的初治成人弥漫性胶质瘤组织样本112例,利用Crispr-Cas12a方法和直接测序法检测样本IDH2-R172K位点突变情况,以直接测序法为金标准。结果112例中,9例IDH2-R172K突变型,103例IDH2-R172K野生型。ROC曲线分析显示,曲线下面积为0.967,约登指数为0.892,最佳截断值为590。以590作为IDH2-R172K突变型和野生型样本的阈值,Crispr-Cas12a法的灵敏性为96.3%(95%CI 81.7%~99.3%),特异性为92.9%(95%CI 89.5%~95.3%)。Crispr-Cas12a法与直接测序法一致性检验Kappa值为0.658,一致性较好。结论Crispr-Cas12a方法能快速、准确地检测成人弥漫性胶质瘤组织样IDH2-R172K突变情况,可作为术中快速检测IDH2-R172K突变的方法。

Objective To investigate the efficacy of Crispr-Cas12a in the detection of R172K single nucleotide polymorphism of isocitrate dehydrogenase 2(IDH2)gene in adult diffuse gliomas.Methods A total of 112 newly diagnosed adult diffuse glioma tissue samples were selected from January 2019 to September 2022,and IDH2-R172K mutations were detected by Crispr-Cas12a and direct sequencing,with direct sequencing as the gold standard.Results Of 112 samples,9 were IDH2-R172K mutant and 103 were IDH2-R172K wild type.ROC curve analysis of Crispr-Cas12a detection results showed that the area under the curve was 0.967,the Jorden index was 0.892,and the best cutoff value was 590.Using 590 as the threshold for IDH2-R172K mutant and wild-type,the sensitivity and specificity of Crispr-Cas12a were 96.3%(95%CI 81.7%~99.3%)and 92.9%(95%CI 89.5%~95.3%),respectively.The Kappa value of Crispr-Cas12a and direct sequencing was 0.658,showing good consistency.Conclusions Crispr-Cas12a can rapidly and accurately detect IDH2-R172K mutation in adult diffuse glioma tissues,and can be used as a method for rapid intraoperative detection of IDH2-R172K mutation.