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基于通用探针的 SNP 基因分型与可视化读取:一种稳健且通用的方法。

Universal probe-based SNP genotyping with visual readout: a robust and versatile method.

机构信息

Key Laboratory of Clinical Laboratory Diagnostics (Chinese Ministry of Education), College of Laboratory Medicine, Chongqing Medical Laboratory Microfluidics and SPRi Engineering Research Center, Chongqing Medical University, Chongqing 400016, P.R. China.

Clinical Laboratories, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, P.R. China.

出版信息

Nanoscale. 2023 Aug 3;15(30):12660-12669. doi: 10.1039/d3nr01950k.

Abstract

Detection of single nucleotide polymorphisms (SNPs) is critical for personalized clinical diagnosis, treatment, and medication. Current clinical detection methods suffer from primer dimerization and require the redesigning of reaction systems for different targets, resulting in a time-consuming and laborious process. Here, we present a robust and versatile method for SNP typing by using tailed primers and universal small molecule probes in combination with a visualized lateral flow assay (LFA). This approach enables not only rapid typing of different targets, but also eliminates the interference of primer dimers and enhances the accuracy and reliability of the results. Our proposed universal assay has been successfully applied to the typing of four SNP loci of clinical samples to verify the accuracy and universality, and the results are consistent with those obtained by Sanger sequencing. Therefore, our study establishes a new universal "typing formula" using nucleic acid tags and small molecule probes that provides a powerful genotyping platform for genetic analysis and molecular diagnostics.

摘要

单核苷酸多态性(SNP)的检测对于个性化临床诊断、治疗和药物使用至关重要。当前的临床检测方法存在引物二聚化问题,并且需要针对不同的靶标重新设计反应体系,导致过程耗时费力。在这里,我们提出了一种使用加尾引物和通用小分子探针结合可视化侧流分析(LFA)进行 SNP 分型的稳健且通用的方法。这种方法不仅能够快速对不同的靶标进行分型,还可以消除引物二聚体的干扰,提高结果的准确性和可靠性。我们提出的通用检测方法已成功应用于临床样本中四个 SNP 位点的分型,以验证准确性和通用性,结果与 Sanger 测序一致。因此,我们的研究建立了一种使用核酸标签和小分子探针的新的通用“分型公式”,为遗传分析和分子诊断提供了强大的基因分型平台。

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