One-step determination of deletion mutation based on loop-mediated isothermal amplification
One-step determination of deletion mutation based on loop-mediated isothermal amplification
复制标题
基于环介导等温扩增的缺失突变一步测定
DOI：
10.1016/j.ab.2020.114087
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发表时间：
2021-01-19
影响因子：
2.9
通讯作者：
Cui, Yali
中科院分区：
文献类型：
--
作者：
Liu, Xiaonan;Zhang, Chao;Cui, Yali
缺失突变已被证明是疾病，特别是癌症发生发展的重要因素。随着精准医学的普及，个体癌症治疗策略强调了开发一种直接有效的缺失突变测定策略的需求。因此，本研究致力于开发一种一步检测方法，以识别具有序列特异性的缺失突变，用于临床实践。利用环介导的等温扩增技术，建立了一种超灵敏、快速的缺失突变检测方法，在强干扰背景下，可在30分钟内准确识别低至30个拷贝或0.1%的目标变异，无需专业操作和复杂的数据解释。作为证明，表皮生长因子受体p.E746-A750del是酪氨酸激酶抑制剂在非小细胞肺癌治疗中易感性的关键因素，该方法已在细胞系和真实临床样本中准确鉴定。通过定制引物组，该方法可以扩展到其他缺失突变体，使其成为一种分子诊断工具，可以很容易地适应于癌症的诊断、治疗和预后的护理点诊断测试场景。
Deletion mutation has been proved as the important factor for occurrence and development of disease, especially those with cancer. With the popularity of precision medicine, the individual cancer therapeutic strategy has highlighted the requirement to develop a straightforward and competent strategy for deletion mutation determination. Hence, the present study is dedicated to develop a one-step assay to identify deletion mutation with sequence specificity for clinical practice. Taking advantage of loop-mediated isothermal amplification, an ultrasensitive and rapid deletion mutation determination method is established, which allow as low as 30 copies or 0.1% target variants under strong interferential background can be accurately distinguished in 30 min dispensing with professional operation and complex data interpretation. As a demonstration, the epidermal growth factor receptor p.E746-A750del, a crucial factor for the susceptibility of tyrosine kinase inhibitor in non-small-cell lung cancer treatment, has been accurately identified by this method with both cell lines and real clinical samples. By tailor-made primer set, this method can be extended for other deletion mutants, making it a molecular diagnostic tool and could be readily adapted for cancer diagnosis, therapy and prognosis in point of care diagnostic test scenario.