摘要目的 建立检测脑胶质瘤患者异柠檬酸脱氢酶1(IDH1)R132H突变的焦磷酸测序方法,并探讨其在临床上的应用性.方法 回顾性纳入中国脑胶质瘤基因组图谱计划(CGGA)数据库中30例脑胶质瘤患者的临床资料和石蜡包埋样本,其中15例为IDH1 R132H野生型,15例为IDH1R132H突变型.采用焦磷酸测序方法检测样本中IDH1 R132H的突变情况,并与免疫组织化学染色结果进行比较分析.结果 30例脑胶质瘤患者焦磷酸测序结果显示,IDH1 132位点CAT所占比例为10％～52％,其中15例IDH1 RI32H野生型样品均＜19％,而其余15例IDH1 R132H突变型样品均＞27％.免疫组织化学染色结果显示,15例IDH1 R132H野生型样品为阴性,其余15例IDH1 R132H突变型为阳性,其中8例为阳性,7例为强阳性.相关性分析结果显示,免疫组织化学染色结果为阴性的样本,焦磷酸测序定量检测的IDH1 R132H突变比例均＜20％ (10％-18％);而免疫组织化学染色结果为阳性的样本,IDH1 R132H突变比例均＞20％(28％～52％);两者间的差异有统计学意义(P＜0.05),且不存在重叠区域.结论 基于焦磷酸测序平台的检测方法可定量分析脑胶质瘤样本IDH1 R132H的突变状态,适合作为临床分子病理学的常规检测.

abstractsObjective To develop a method based on pyrosequencing that can be used to accurately and quantitatively detect the IDH1 R132H mutation of gliomas and to evaluate the clinical application of that method.Methods Thirty cases of formalin-fixed and parrffin-embedded (FFPE) samples of gliomas and clinical database were retrospectively obtained from the Chinese Glioma Genome Atlas (CGGA).There are 15 IDH1 wild type and 15 IDH1 R132H mutant samples.The pyrosequencing results were compared with the findings of immunohistochemistry.Results The pyrosequencing results indicated that the percentage of CAT in glioma sample ranged from 10％ to 52％ in all 30 cases.The percentage of CAT in IDH1 R132H mutant glioma was less than 19％,while the percentage of CAT in IDH1 R132H wildtype glioma was more than 27％.Immunohistochemistry results indicated negative staining in 15 samples and positive staining in the other 15 samples including 8 positive staining and 7 strong positive staining.Further statistical analysis showed that the quantitative IDH1 R132H mutant ratios of pyrosequencing were significantly different between immunohistochemistry-negative and immunohistochemistry-positive glioma FFPE samples,and the former was below 20％ (10％-18％),while the latter was above 20％ (28％-52％).Conclusion The quantitative IDH1 R132H mutant detection method based on pyrosequencing could accurately detect the IDH1 R132H mutant level in FFPE glioma samples,which seems suitable for routine molecular pathological practice.