褪色HE染色切片ALK基因融合检测可行性探索  

Feasibility exploration of ALK gene fusion detection in faded HE-stained sections

作　　者：李钦丽 张继伟[1] Li Qinli;Zhang Jiwei(Department of Pathology,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China;Department of Clinical Laboratory,Dongxihu District People’s Hospital,Wuhan 430040,China)

机构地区：[1]华中科技大学同济医学院附属协和医院病理科,武汉430022 [2]武汉市东西湖区人民医院检验科,武汉430040

出　　处：《中国组织化学与细胞化学杂志》2023年第4期396-401,共6页Chinese Journal of Histochemistry and Cytochemistry

摘　　要：目的 探索利用HE染色切片褪色样本进行ALK基因融合检测的可行性。方法 选取3例ALK基因融合阳性的福尔马林固定石蜡包埋(formalin fixed and paraffin embedded,FFPE)组织样本的HE染色切片,分别经盐酸乙醇褪色法和高锰酸钾草酸氧化漂白法褪色后,提取RNA,采用荧光定量RT-PCR(qRT-PCR)技术,检测ALK基因融合。结果 从经盐酸乙醇褪色法或高锰酸钾草酸氧化漂白法褪色的HE染色切片中提取的RNA的纯度与从对照切片中提取者无显著差异,但其浓度较从对照切片中提取者显著降低。在模板量相同的条件下,经盐酸乙醇褪色的HE染色切片与对照组切片均检测到明确的ALK基因融合阳性,而经高锰酸钾草酸褪色的HE染色切片无任何曲线升起。从经盐酸乙醇褪色的HE染色切片中提取的RNA的完整性与从对照切片中提取的RNA的完整性无差异,但显著高于从经高锰酸钾草酸褪色的HE染色切片中提取者。为保证研究的准确性,本研究采用免疫组织化学与荧光原位杂交法对选取的ALK基因融合阳性样本进行验证,结果发现所选样本均为明确阳性。结论 HE染色切片经盐酸乙醇褪色后提取的RNA可以用于ALK基因融合检测。Objective To explore the feasibility of ALK gene fusion detection using the faded HE-stained sections.Methods The HE-stained sections of three formalin fixed and paraffin embedded tissue samples with positive ALK gene fusion were selected.After fading by hydrochloric acid ethanol fading method and potassium permanganate oxalic acid oxidation bleaching method,respectively,RNA was extracted,and fluorescence quantitative RT-PCR(qRT-PCR)was used to detect ALK gene fusion.Results The purity of RNA extracted from HE-stained sections faded by hydrochloric acid ethanol fading method or by potassium permanganate oxalic acid oxidation bleaching method had no significant difference from that of RNA extracted from control sections,but its concentration was significantly lower than that of RNA extracted from control sections.Under the condition of the same amount of template,positive ALK gene fusion was detected in both HE-stained sections faded by hydrochloric acid ethanol and control group sections,while no curve rose in HE-stained sections faded by potassium permanganate oxalic acid.The integrity of RNA extracted from HE-stained sections faded by hydrochloric acid ethanol was no different from that of RNA extracted from control sections,but significantly higher than that of RNA extracted from HE stained sections faded by potassium permanganate oxalic acid.In order to ensure the accuracy of the study,immunohistochemistry and fluorescence in situ hybridization were used to verify the selected ALK gene fusion positive samples,and the results showed that the selected samples were definitely positive.Conclusion The RNA extracted from HE-stained sections faded by hydrochloric ethanol can be used for fusion detection of ALK gene.

关 键 词：HE染色切片 褪色 ALK基因融合 基因检测 

分 类 号：R734.2[医药卫生—肿瘤]