Dual detection of Bupleurum scorzonerifolium Willd. and Bupleurum chinense DC. using proofman–LMTIA method

IF 5.2 2区农林科学 Q1 AGRICULTURE, MULTIDISCIPLINARY

Chemical and Biological Technologies in Agriculture Pub Date : 2024-08-02 DOI:10.1186/s40538-024-00637-2

Jinxin Liu, Yue Wang, Tingting Li, Kunpeng Huang, Chunmei Song, Pengfei Cui, Bailing Yin, Juntao Sun, Fugang Xiao, Deguo Wang

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Abstract

A new rapid dual detection method was established to distinguish Bupleurum scorzonerifolium Willd. (BS) and Bupleurum chinense DC. (BC) simultaneously using the proofreading enzyme-mediated probe cleavage coupled with ladder-shape melting temperature isothermal amplification (proofman–LMTIA). The internal transcribed spacer (ITS) sequences of BS and BC were selected as targets for designs of proofman–LMTIA primers and proofman fluorescence probes labeled with FAM or JOE. The reaction temperature optimization, repeatability and reliability assessment, specificity assessment and sensitivity assessment of the proofman–LMTIA performed for dual detection of BS and BC and its application. The results showed that the optimal reaction temperature of the proofman–LMTIA method was at 63℃, which had strongly specificity, repeatability and reliability, as well as sensitivity, and the detection was completed within 20 min with a detection sensitivity of 1 pg/μL. The proofman–LMTIA method realized dual rapid detection of BS and BC, which showed a strong practical value. The 4 kinds were BC, 1 kind was BS, and 2 kinds were counterfeit in the detection of 7 kinds of BS or BC samples from different habitats. Our study successfully established a new approach for dual rapid detection of BS and BC using the proofman–LMTIA, which will provide an effective technique or method for the authenticity detection of authentic Chinese medicinal materials and present a very important practical significance.

Graphical Abstract

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使用 proofman-LMTIA 方法对 Bupleurum scorzonerifolium Willd.和 Bupleurum chinense DC.进行双重检测

利用校对酶介导的探针裂解结合梯形熔融温度等温扩增（proofman-LMTIA）技术，建立了一种新的快速双重检测方法，可同时区分Bupleurum scorzonerifolium Willd.（BS）和Bupleurum chinense DC.(利用校对酶介导的探针裂解结合梯形熔融温度等温扩增（proofman-LMTIA）技术，建立了一种新的快速双重检测方法，可同时区分Bupleurum scorzonerifolium Willium（BS）和Bupleurum chinense DC.（BC）。以 BS 和 BC 的内部转录间隔序列（ITS）为目标，设计了校对人-LMTIA 引物和用 FAM 或 JOE 标记的校对人荧光探针。对proofman-LMTIA进行了反应温度优化、重复性和可靠性评估、特异性评估和灵敏度评估，用于BS和BC的双重检测及其应用。结果表明，proofman-LMTIA方法的最佳反应温度为63℃，具有较强的特异性、重复性、可靠性和灵敏度，可在20 min内完成检测，检测灵敏度为1 pg/μL。proofman-LMTIA方法实现了BS和BC的双重快速检测，具有很强的实用价值。在对来自不同生境的 7 种 BS 或 BC 样品的检测中，4 种为 BC，1 种为 BS，2 种为假货。我们的研究成功地建立了利用放样器-LMTIA进行BS和BC双重快速检测的新方法，这将为地道中药材的真伪检测提供一种有效的技术或方法，具有非常重要的现实意义。

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来源期刊

Chemical and Biological Technologies in Agriculture Biochemistry, Genetics and Molecular Biology-Biotechnology

CiteScore

6.80

自引率

3.00%

发文量

审稿时长

15 weeks

期刊介绍： Chemical and Biological Technologies in Agriculture is an international, interdisciplinary, peer-reviewed forum for the advancement and application to all fields of agriculture of modern chemical, biochemical and molecular technologies. The scope of this journal includes chemical and biochemical processes aimed to increase sustainable agricultural and food production, the evaluation of quality and origin of raw primary products and their transformation into foods and chemicals, as well as environmental monitoring and remediation. Of special interest are the effects of chemical and biochemical technologies, also at the nano and supramolecular scale, on the relationships between soil, plants, microorganisms and their environment, with the help of modern bioinformatics. Another special focus is the use of modern bioorganic and biological chemistry to develop new technologies for plant nutrition and bio-stimulation, advancement of biorefineries from biomasses, safe and traceable food products, carbon storage in soil and plants and restoration of contaminated soils to agriculture. This journal presents the first opportunity to bring together researchers from a wide number of disciplines within the agricultural chemical and biological sciences, from both industry and academia. The principle aim of Chemical and Biological Technologies in Agriculture is to allow the exchange of the most advanced chemical and biochemical knowledge to develop technologies which address one of the most pressing challenges of our times - sustaining a growing world population. Chemical and Biological Technologies in Agriculture publishes original research articles, short letters and invited reviews. Articles from scientists in industry, academia as well as private research institutes, non-governmental and environmental organizations are encouraged.