Long Jinyan, Chen Mengqi, Yu Yang, Wu Qiaomin, Yang Xiaolan
Key Laboratory of Medical Laboratory Diagnostics of the Education Ministry, College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Key Laboratory of Medical Laboratory Diagnostics of the Education Ministry, College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Anal Chim Acta. 2023 Oct 2;1276:341617. doi: 10.1016/j.aca.2023.341617. Epub 2023 Jul 23.
The detection of single nucleotide variants (SNVs) is important for the diagnosis and treatment of cancer. To date, researchers have devised several methods to detect SNVs, but most of them are complex and time-consuming. To improve SNVs detection specificity and sensitivity, we developed a triple-recognition strategy, which facilitates aligner-mediated cleavage-triggered exponential amplification (Trec-AMC-EXPAR) for the rapid, specific, and one-pot detection of SNV. Under optimized conditions, Trec-AMC-EXPAR detected two clinically significant SNVs, PIK3CA and EGFR L858R within 80 min, with a reliable detection of 0.1% SNV in the wide type, which is lower than that of allele-specific PCR (AS-PCR) for detecting SNV. Finally, by spiking into normal human serum samples, mutants mixed with the wild-type targets in different ratios were analyzed, resulting in the relative standard deviation (RSD) of recovery ratios <3%. The findings suggested the potential application of Trec-AMC-EXPAR in clinical disease diagnosis. In summary, the proposed Trec-AMC-EXPAR technique provides a novel fast and convenient method for one-pot detection of SNV with high sensitivity and specificity.
单核苷酸变异（SNV）的检测对于癌症的诊断和治疗至关重要。迄今为止，研究人员已经设计出多种检测SNV的方法，但其中大多数方法复杂且耗时。为了提高SNV检测的特异性和灵敏度，我们开发了一种三重识别策略，即促进比对介导的切割触发指数扩增（Trec-AMC-EXPAR），用于快速、特异性和一锅法检测SNV。在优化条件下，Trec-AMC-EXPAR在80分钟内检测到两个具有临床意义的SNV，即PIK3CA和EGFR L858R，在野生型中可靠检测限为0.1%的SNV，低于等位基因特异性PCR（AS-PCR）检测SNV的检测限。最后，通过加入正常人血清样本，分析了不同比例混合的突变体与野生型靶标的情况，回收率的相对标准偏差（RSD）<3%。这些发现表明Trec-AMC-EXPAR在临床疾病诊断中的潜在应用。总之，所提出的Trec-AMC-EXPAR技术为一锅法检测SNV提供了一种新颖、快速且便捷的方法，具有高灵敏度和特异性。
