基于CRISPR-Cas12a检测成人型弥漫性胶质瘤MGMT启动子甲基化  

Detection of MGMT promoter methylation based on CRISPR-Cas12a method in adult diffuse glioma

作　　者：冯哲斌 孔东生 许军鹏 武其嘉 张宇 余新光 冯世宇 FENG Zhebin;KONG Dongsheng;XU JunPeng(Department of Neurosurgery,the First Medical Center of PLA General Hospital,Beijing 100089,China)

机构地区：[1]中国人民解放军总医院第一医学中心神经外科医学部,北京100089

出　　处：《临床神经外科杂志》2023年第6期664-670,共7页Journal of Clinical Neurosurgery

基　　金：军队装备综合研究和技术基础项目(LB20201A060004)。

摘　　要：目的应用成簇规律间隔短回文重复序列相关12a(CRISPR-Cas12a)方法检测成人型弥漫性胶质瘤O 6-甲基鸟嘌呤-DNA甲基转移酶(MGMT)启动子甲基化情况,并初步评价该方法的检测效能及其与高通量测序结果的一致性。方法选择2022年3月—2022年4月解放军总医院第一医学中心神经外科手术切除的初治成人型弥漫性胶质瘤组织样本,分别利用CRISPR-Cas12a方法和高通量测序对样本MGMT启动子甲基化情况进行检测。计算CRISPR-Cas12a方法的灵敏度、特异性等效能指标,并利用配对χ2检验(McNemar检验)及Kappa一致性检验分析CRISPR-Cas12a方法与高通量测序结果的一致性。结果共纳入32例成人型弥漫性胶质瘤组织样本。CRISPR-Cas12a方法检测成人型弥漫性胶质瘤组织样本MGMT启动子甲基化序列的灵敏度为98.4%,特异性为90.9%;检测MGMT启动子非甲基化序列的灵敏度为100%,特异性为90.5%。两种CRISPR-Cas12a方法的检测结果与高通量测序结果一致性均较好。结论Crispr-Cas12a方法能较准确地对成人型弥漫性胶质瘤组织样本MGMT启动子甲基化情况进行检测,有望成为一种新兴的检测MGMT启动子甲基化情况的方法。Objective Using the CRISPR-Cas12a method to detect O 6-methylguanine-DNA methyltransferase(MGMT)promoter methylation in adult diffuse glioma,and initially evaluating the efficacy of this method and the consistency with the results of high-throughput sequencing method.Methods The tissue samples of newly diagnosed adult diffuse glioma removed in Department of Neurosurgery,the First Medical Center of PLA General Hospital from March 2022 to April 2022 were selected for MGMT promoter methylation detection using the CRISPR-Cas12a method and high-throughput sequencing method,respectively.The sensitivity and specificity of the CRISPR-Cas12a method were calculated,and the consistency between the CRISPR-Cas12a method and the high-throughput sequencing method was analyzed by using the paired chi-square test(McNemar test)and the Kappa consistency test.Results A total of 32 adult diffuse glioma tissue samples were included.The Crispr-Cas12a method had a sensitivity of 98.4%,specificity of 90.9%in detecting MGMT promoter methylation sequences in adult diffuse glioma tissue samples.The Crispr-Cas12a method had a sensitivity of 100%,specificity of 90.5%in detecting MGMT promoter unmethylation sequences.The results of both two CRISPR-Cas12a methods showed good consistency with the results of high-throughput sequencing method.Conclusion The Crispr-Cas12a method is able to relatively accurately detect MGMT promoter methylation in adult diffuse glioma tissue samples,and it is expected to be an emerging method for detecting the status of MGMT promoter methylation.

关 键 词：CRISPR-Cas12a 成人型弥漫性胶质瘤 MGMT启动子甲基化 NGS测序 MGMT蛋白 

分 类 号：R739.41[医药卫生—肿瘤]