孙雪睿,
王玉,
高佳林,
李梅^,
大连医科大学附属第二医院 科研中心, 辽宁 大连 116027

基金项目: 国家自然科学基金面上项目（81972022）；国家自然科学青年基金项目（81501832）；辽宁省自然科学基金项目（2019-MS-087）


详细信息 作者简介: 孙雪睿(1994-), 女, 硕士研究生




通讯作者: 李梅, 副主任医师。E-mail:li.mei@dmu.edu.cn 中图分类号: R331


摘要:目的探讨饱和荧光染料LCGreen Plus与EvaGreen在HRM法基因变异检测中的差异。方法选取34例课题组保存的DNA标本（EGFR第18-21外显子突变情况已知），以双盲法分别应用LCGreen Plus与EvaGreen染料HRM法在Rotor-Gene Q平台进行上述片段基因变异检测，由两位拥有HRM分析基础的人员进行结果判定，比较两种染料基因检测的准确率并分析熔解曲线特征。结果LCGreen Plus和EvaGreen检测上述4个基因片段的准确率、灵敏性和特异性分别为100%（136/136）和90.4%（123/136）、100%（69/69）和81.1%（56/69）、100%（67/67）和100%（67/67）。LCGreen Plus和EvaGreen标记的熔解曲线特征不同。结论了解饱和双链DNA染料标记的熔解曲线特征，选用合适判读方法可能有助于减少假阴性结果的产生。
关键词: LCGreen Plus/
EvaGreen/
高分辨熔解曲线


Abstract:ObjectiveTo compare saturated fluorescent dyes LCGreen Plus with EvaGreen in detecting EGFR mutation with HRM.MethodsSelect 34 DNA samples with known genotype in EGFR exon 18-21. With double blind method, all these samples were detected genotype of the four fragments in HRM with LCGreen Plus and EvaGreen, separately, on Rotor-Gene Q. Results were called independently by two researchers with HRM analysis experience. Accuracies, sensitivities and specificities of mutation detection were calculated and melting curve characters were analyzed.ResultsFor HRM with LCGreen Plus and EvaGreen, the accuracy, sensitivity, specificity were 100% (136/136) and 90.4% (123/136), 100% (69/69) and 81.1% (56/69), 100% (67/67) and 100% (67/67). Melting curves marked by LCGreen Plus and EvaGreen appear different characteristics.ConclusionIt is important to know HRM curve characteristics with a specific dye to call genotype correctly.
Keywords:LCGreen Plus/
EvaGreen/
HRM

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https://journal.dmu.edu.cn/data/article/export-pdf?id=dlykdxxb_20200502