A recombinase polymerase amplification-SYBR Green I assay for the rapid and visual detection of Brucella.

IF 2.4 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Folia microbiologica Pub Date : 2024-08-01 Epub Date: 2023-12-02 DOI:10.1007/s12223-023-01115-2

Jiang Chang, Nan Wang, Jun-Peng Zhan, Shi-Jun Zhang, De-Ying Zou, Feng Li, Ying Zhang, Yan-Song Li, Pan Hu, Shi-Ying Lu, Zeng-Shan Liu, Hong-Lin Ren

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Abstract

Brucellosis is a zoonosis caused by Brucella, which poses a great threat to human health and animal husbandry. Pathogen surveillance is an important measure to prevent brucellosis, but the traditional method is time-consuming and not suitable for field applications. In this study, a recombinase polymerase amplification-SYBR Green I (RPAS) assay was developed for the rapid and visualized detection of Brucella in the field by targeting BCSP31 gene, a conserved marker. The method was highly specific without any cross-reactivity with other common bacteria and its detection limit was 2.14 × 10⁴ CFU/mL or g of Brucella at 40 °C for 20 min. It obviates the need for costly instrumentation and exhibits robustness towards background interference in serum, meat, and milk samples. In summary, the RPAS assay is a rapid, visually intuitive, and user-friendly detection that is highly suitable for use in resource-limited settings. Its simplicity and ease of use enable swift on-site detection of Brucella, thereby facilitating timely implementation of preventive measures.

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一种重组酶聚合酶扩增- sybr Green I检测布鲁氏菌的方法。

布鲁氏菌病是由布鲁氏菌引起的人畜共患病，对人类健康和畜牧业造成极大威胁。病原菌监测是预防布鲁氏菌病的重要措施，但传统方法耗时长，不适合现场应用。本研究以布鲁氏菌的保守标记BCSP31基因为靶点，建立了一种重组酶聚合酶扩增- sybr Green I (RPAS)方法，用于布鲁氏菌的野外快速、可视化检测。该方法具有高度特异性，与其他常见细菌无交叉反应性，在40°C、20 min条件下，布鲁氏菌的检出限为2.14 × 104 CFU/mL或g。该方法不需要昂贵的仪器，对血清、肉类和牛奶样品的背景干扰具有鲁棒性。总之，RPAS分析法是一种快速、直观、用户友好的检测方法，非常适合在资源有限的环境中使用。它简单易用，能够在现场迅速检测布鲁氏菌，从而促进及时实施预防措施。

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

Folia microbiologica 工程技术-生物工程与应用微生物

CiteScore

5.80

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： Unlike journals which specialize ever more narrowly, Folia Microbiologica (FM) takes an open approach that spans general, soil, medical and industrial microbiology, plus some branches of immunology. This English-language journal publishes original papers, reviews and mini-reviews, short communications and book reviews. The coverage includes cutting-edge methods and promising new topics, as well as studies using established methods that exhibit promise in practical applications such as medicine, animal husbandry and more. The coverage of FM is expanding beyond Central and Eastern Europe, with a growing proportion of its contents contributed by international authors.