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使用钳制肽核酸（PNA）和PNA检测探针同时对多个体细胞突变进行基因分型。

Simultaneous genotyping of multiple somatic mutations by using a clamping PNA and PNA detection probes.

作者信息

Kim Yong-Tae, Kim Jin Woo, Kim Sung Kee, Joe Goon Ho, Hong In Seok

机构信息

Research Institute, Panagene Inc. 54 Techno10-ro, Yuseong-gu, Daejeon, 305-510 (Republic of Korea).

出版信息

Chembiochem. 2015 Jan 19;16(2):209-13. doi: 10.1002/cbic.201402640. Epub 2014 Dec 22.

DOI:10.1002/cbic.201402640
PMID:25534284
Abstract

It has been very difficult to detect and quantify multiple somatic mutations simultaneously in single-tube qPCR. Here, a novel method for simultaneous detection of multiple mutations and a melting curve analysis was developed by using clamping PNA and detection PNA probes. Each PNA probe was designed to have a specific melting temperature by the introduction of γ-PNA monomer. This technique was successfully applied to the detection of six genotypes in two different mutations of K-RAS at the same time. Such simultaneous analysis of an amplified curve and a melting curve in qPCR can be widely used for the early diagnosis of cancer and determining the prognosis of drug treatments.

摘要

在单管定量聚合酶链反应（qPCR）中同时检测和定量多个体细胞突变一直非常困难。在此，通过使用钳制肽核酸（PNA）和检测PNA探针，开发了一种同时检测多个突变和熔解曲线分析的新方法。通过引入γ-PNA单体，每个PNA探针被设计成具有特定的熔解温度。该技术已成功应用于同时检测K-RAS两个不同突变中的六种基因型。qPCR中扩增曲线和熔解曲线的这种同步分析可广泛用于癌症的早期诊断和确定药物治疗的预后。

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