聚羟基丙烯酸和Van-clear在qRT-PCR法检测非小细胞肺癌EGFR基因突变中的应用研究

发布时间：2018-06-14 17:14

本文选题：聚羟基丙烯酸 + 环保透明脱蜡液Van-clear　；参考：《西安交通大学学报(医学版)》2017年05期

【摘要】：目的比较环保固定液聚羟基丙烯酸、环保透明脱蜡液Van-clear单独或联合替代传统固定液4%中性缓冲甲醛、传统透明脱蜡液二甲苯应用于实时荧光定量PCR(qRT-PCR)法,检测非小细胞肺癌(NSCLC)表皮生长因子受体(EGFR)基因突变率的敏感性、特异性及符合率的差异,评估qRT-PCR法及其环保技术平台的临床应用价值。方法选取中山市博爱医院、中山市中医院2013年5月至2016年3月期间手术切除的原发性NSCLC标本91例,同一肿瘤病变部位切取5个样本,采用随机数字表法分为A、B、C、D、E。A组使用4%中性缓冲甲醛固定、二甲苯透明脱蜡制作切片、直接测序法检测EGFR基因突变;B组使用4%中性缓冲甲醛固定、二甲苯透明脱蜡制作切片、qRT-PCR法检测EGFR基因突变;C组使用聚羟基丙烯酸固定、二甲苯透明脱蜡制作切片、qRT-PCR法检测EGFR基因突变;D组使用4%中性缓冲甲醛固定、Van-clear透明脱蜡制作切片、qRT-PCR法检测EGFR基因突变;E组使用聚羟基丙烯酸固定、Van-clear透明脱蜡制作切片、qRT-PCR法检测EGFR基因突变。分别测定5组NSCLC中EGFR基因第18、19、20、21号外显子的突变情况。结果 (1)B、C、D、E组与A组比较,EGFR基因发生突变人数百分率、基因靶位点突变率的差异均无统计学意义(P0.05);(2)B、C、D、E组与A组比较,EGFR靶位点突变检测结果的灵敏度好、特异度强、符合率高。结论聚羟基丙烯酸、环保透明脱蜡液Van-clear单独或联合替代传统试剂,应用于qRT-PCR法检测NSCLC中EGFR基因突变,与以传统试剂应用于直接测序法的检测结果一致性好,有在临床推广应用的意义和价值。
[Abstract]:Objective to compare the use of polyhydroxy acrylic acid (PAA), environmental transparent dewaxing solution (Van-clear) to replace 4% neutral buffered formaldehyde in traditional fixative solution, and the application of xylene in real-time fluorescence quantitative PCRQRT-PCRmethod. The sensitivity, specificity and coincidence rate of EGF receptor EGFR gene mutation in NSCLC were detected. The clinical application value of qRT-PCR and its environmental protection technology platform was evaluated. Methods 91 cases of primary NSCLC from Zhongshan Boai Hospital and Zhongshan Hospital of traditional Chinese Medicine from May 2013 to March 2016 were selected. The method of random digital table was used to divide the control group into 4% neutral buffered formaldehyde fixation, xylene transparent dewaxing section and 4% neutral buffered formaldehyde fixation to detect EGFR gene mutation in group B by direct sequencing. The EGFR gene mutation was detected by qRT-PCR in transparent dewaxing section of xylene. Group C was immobilized with polyhydroxyacrylic acid. Detection of EGFR gene mutation by qRT-PCR in group D using 4% neutral buffer formaldehyde fixed Van-clear transparent dewaxing section qRT-PCR was used to detect EGFR gene mutation. Group E used polyhydroxyacrylic acid to fix Van-clear transparent dewaxing. EGFR gene mutation was detected by qRT-PCR. The mutation of exon 18 of EGFR gene in 5 groups of NSCLC was determined. Results there was no significant difference in the mutation rate of EGFR gene between group E and group A, and there was no significant difference between group E and group A in the sensitivity, specificity and coincidence rate of detection of EGFR target mutation in group E and group A. Conclusion Polyhydroxy acrylic acid, environmental transparent dewaxing solution Van-clear is used to detect EGFR gene mutation in NSCLC by using qRT-PCR method, which is consistent with the results of direct sequencing with traditional reagent. Has the significance and the value in the clinical popularizing application.
【作者单位】：南方医科大学附属中山博爱医院病理科;广东医科大学广东省医学分子诊断重点实验室;广州中医药大学附属中山医院病理科;
【基金】：国家自然科学基金资助项目(No.81101843) 广东省医学科学技术研究基金项目(No.A2017321) 广东省中山市卫生和计划生育局医学科研立项课题(No.2014J128)~~
【分类号】：R734.2

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