Experimental study on the detection of Gastrodia elata by enzymatic recombinase amplification and immunochromatography

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry Pub Date : 2024-07-14 DOI:10.1016/j.ab.2024.115618

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引用次数: 0

Abstract

Objective

The objective of this research is to develop two methodologies, Enzymatic recombinase amplification (ERA) and Polymerase Chain Reaction (PCR) coupled with Lateral Flow Dipstick (LFD), for the swift authentication of Gastrodia elata.

Methodology

Primers and nfo probes for the ERA of Gastrodia elata were developed based on the ITS2 genome sequences of Gastrodia elata and its counterfeits. Specific primers for the PCR analysis of Gastrodia elata were generated using the NCBI (National Center for Biotechnology Information) online platform. Through experimental validation, the optimal reaction system and conditions for both methodologies were established, and their efficacy was assessed.

Results

The methodologies developed herein are applicable for the targeted analysis of the medicinal species, Gastrodia elata. The sensitivity of the ERA-LFD detection method matched that of the conventional PCR-LFD approach, recorded at 1 ng μL⁻¹. Consistency was observed in the results across three replicates of visualization test strips for both techniques. Upon evaluation, both the PCR-LFD and ERA-LFD methods demonstrated a total compliance rate of 100 %.

Conclusion

The ERA-LFD and PCR-LFD methods facilitate reduced detection times and offer visual results. These techniques are particularly effective for on-site detection and quality control in the authentication of Gastrodia elata within traditional Chinese medicine markets and at the primary level of healthcare provision.

Abstract Image

查看原文本刊更多论文

利用酶重组酶等温扩增和免疫层析技术检测 G.elata。

研究目的本研究旨在开发两种方法，即酶重组酶扩增法（ERA）和聚合酶链式反应法（PCR）以及侧流浸量尺（LFD），用于快速鉴定天麻：根据天麻及其仿冒品的 ITS2 基因组序列，开发了用于天麻 ERA 的引物和 nfo 探针。利用 NCBI（美国国家生物技术信息中心）在线平台生成了用于对天麻进行 PCR 分析的特异性引物。通过实验验证，确定了两种方法的最佳反应体系和条件，并评估了其有效性：结果：本文所开发的方法适用于药用植物天麻的靶向分析。ERA-LFD检测方法的灵敏度与传统的PCR-LFD方法相当，均为1 ng-μL-1。两种技术在三个重复的可视化测试条上的结果一致。经评估，PCR-LFD 和 ERA-LFD 方法的总符合率均为 100%：结论：ERA-LFD 和 PCR-LFD 方法有助于缩短检测时间并提供可视化结果。结论：ERA-LFD 和 PCR-LFD 方法可缩短检测时间，并提供直观结果。这些技术对传统中药市场和基层医疗机构的天麻鉴定现场检测和质量控制尤为有效。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Analytical biochemistry 生物-分析化学

CiteScore

5.70

自引率

0.00%

发文量

283

审稿时长

44 days

期刊介绍： The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field. The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology. The journal has been particularly active in: -Analytical techniques for biological molecules- Aptamer selection and utilization- Biosensors- Chromatography- Cloning, sequencing and mutagenesis- Electrochemical methods- Electrophoresis- Enzyme characterization methods- Immunological approaches- Mass spectrometry of proteins and nucleic acids- Metabolomics- Nano level techniques- Optical spectroscopy in all its forms. The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.