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A new simplified sequence-dependent loop-mediated isothermal amplification (LAMP) detection method

文献类型： 外文期刊

作者： Chen, Yanju 1 ; Zhu, Yuanyuan 1 ; Du, Jungang 1 ; Peng, Cheng 3 ; Wang, Xiaofu 3 ; Wu, Jian 1 ; Zhou, Qingli 4 ; Chen, Huan 5 ; Xu, Junfeng 3 ;

作者机构： 1.Zhejiang Univ, Coll Biosyst Engn & Food Sci, Hangzhou 311215, Peoples R China

2.Zhejiang Univ, ZJU Hangzhou Global Sci & Technol Innovat Ctr, Hangzhou 311215, Peoples R China

3.Zhejiang Acad Agr Sci, Key Lab Traceabil Agr Genet Modified Organisms, Minist Agr & Rural Affairs, Hangzhou 310021, Peoples R China

4.Zhejiang Univ, Affiliated Hosp 4, Sch Med, Yiwu 322000, Peoples R China

5.Hangzhou Digital Micro Biotech Co Ltd, Hangzhou 311215, Peoples R China

关键词： Isothermal amplification; LAMP; Sequence-dependent probe; Multiplexed detection; Diagnostics

期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY （ 影响因子：4.3； 五年影响因子：4.0 ）

ISSN： 1618-2642

年卷期： 2024 年

页码：

收录情况： SCI

摘要： Fluorescence dye-based loop-mediated isothermal amplification (LAMP) is a sensitive nucleic acid detection method, but is limited to single-plex detection and may yield non-specific signals. In this study, we propose a bifunctional probe-based real-time LAMP amplification method for single-plexed or multiplexed detection. The bifunctional probe is derived by modifying the 5 ' end of the fluorophore and an internal quencher on one of the LAMP primers; therefore, it can simultaneously be involved in the LAMP process and signal amplification. The fluorescence intensity undergoes a cumulative exponential increase during the incorporation of the bifunctional probe into double-stranded DNA amplicons. The bifunctional probe-based LAMP method is simplified and cost-effective, as the primer design and experimental operations align entirely with the ordinary LAMP. Different from other current probe-based methods, this method does not require additional enzymes, sequences, or special probe structures. Also, it is 10 min faster than several other probe-based LAMP methods. The bifunctional probe-based LAMP method allows the simultaneous detection of the target Vibrio parahaemolyticus DNA and the internal amplification control in a one-pot reaction, demonstrating its potential for multiplexed detection.

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