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Multiplex Detection of Single Nucleotide Polymorphisms by Liquid Chromatography for Nonsmall Cell Lung Cancer Staging
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-01-08 , DOI: 10.1021/acs.analchem.3c03659 Chang Song 1 , Ziyu Ma 1 , Mengyu Zhang 1 , Chang Liu 2 , Sheng Tang 1 , Jinghui Zhang 1 , Juan Song 1 , Hui Yu 3 , Hian Kee Lee 1, 4 , Wei Shen 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-01-08 , DOI: 10.1021/acs.analchem.3c03659 Chang Song 1 , Ziyu Ma 1 , Mengyu Zhang 1 , Chang Liu 2 , Sheng Tang 1 , Jinghui Zhang 1 , Juan Song 1 , Hui Yu 3 , Hian Kee Lee 1, 4 , Wei Shen 1
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In this work, an integrated strategy with excellent accuracy and high throughput is proposed for the precise indication of single nucleotide polymorphism (SNP) in nonsmall cell lung cancer diseases. Two types of point mutations (L858R and T790M) and the corresponding wild types could be identified together in a single high-performance liquid chromatographic run. Signal amplification was achieved through a series of enzyme ligation, primer extension, and enzyme cleavage strategies, and a large number of DNA probes with different fluorescence signals were finally generated. The factors affecting the spatiotemporal separation efficiency of four DNA probes were systematically investigated. The limits of detection of wild types (WTs) or mutant types (MTs) abbreviated as L858R-MT, L858R-WT, T790M-MT, and T790M-WT were 26, 24, 19, and 22 aM, respectively. In addition, the levels of mutant types and wild types in the serum of 40 nonsmall cell lung cancer patients at different stages were detected using the method, and the correlation between the mutation ratios and cancer stages was preliminarily verified. The proposed highly selective and sensitive method may serve as an alternative approach for early diagnosis and staging of nonsmall cell lung cancer.
中文翻译：
液相色谱多重检测单核苷酸多态性用于非小细胞肺癌分期
在这项工作中，提出了一种具有出色准确性和高通量的综合策略，用于精确指示非小细胞肺癌疾病中的单核苷酸多态性（SNP）。两种类型的点突变（L858R 和 T790M）和相应的野生型可以在一次高效液相色谱运行中一起鉴定。通过一系列酶连接、引物延伸、酶切策略实现信号放大，最终产生大量具有不同荧光信号的DNA探针。系统研究了四种DNA探针时空分离效率的影响因素。野生型 (WT) 或突变型 (MT) 缩写为 L858R-MT、L858R-WT、T790M-MT 和 T790M-WT 的检测限分别为 26、24、19 和 22 aM。此外，利用该方法检测了40例不同分期非小细胞肺癌患者血清中突变型和野生型的水平，初步验证了突变比例与癌症分期的相关性。所提出的高度选择性和敏感性的方法可以作为非小细胞肺癌早期诊断和分期的替代方法。
更新日期：2024-01-08
中文翻译：
液相色谱多重检测单核苷酸多态性用于非小细胞肺癌分期
在这项工作中，提出了一种具有出色准确性和高通量的综合策略，用于精确指示非小细胞肺癌疾病中的单核苷酸多态性（SNP）。两种类型的点突变（L858R 和 T790M）和相应的野生型可以在一次高效液相色谱运行中一起鉴定。通过一系列酶连接、引物延伸、酶切策略实现信号放大，最终产生大量具有不同荧光信号的DNA探针。系统研究了四种DNA探针时空分离效率的影响因素。野生型 (WT) 或突变型 (MT) 缩写为 L858R-MT、L858R-WT、T790M-MT 和 T790M-WT 的检测限分别为 26、24、19 和 22 aM。此外，利用该方法检测了40例不同分期非小细胞肺癌患者血清中突变型和野生型的水平，初步验证了突变比例与癌症分期的相关性。所提出的高度选择性和敏感性的方法可以作为非小细胞肺癌早期诊断和分期的替代方法。
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