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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Funding
This work was supported by grants from the National Key Research and Development Program of China (No. 2021YFC2600202), the Science and Technology Development Project of Jilin Province, China (No. YDZJ202101ZYTS124), the Science and Technology Project of Science and Technology Development Program of Changchun City (No. 21ZGN34), and the Fundamental Research Funds for the Central Universities (No. 45121031B007).
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Jiang Chang participated in methodology, writing and editing. Nan Wang participated in validation. Jun-Peng Zhan participated in writing original draft. Shi-Jun Zhang participated in methodology. De-Ying Zou participated in visualization. Feng Li participated in resources; Ying Zhang participated in investigation. Yan-Song Li participated in supervision. Pan Hu participated in review and editing. Shi-Ying Lu participated in validation. Zeng-Shan Liu participated in formal analysis. Hong-Lin Ren participated in conceptualization, funding acquisition, and project administration. Jiang Chang, Nan Wang, Jun-Peng Zhan, and Shi-Jun Zhang contributed equally in this work.
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Chang, J., Wang, N., Zhan, JP. et al. A recombinase polymerase amplification–SYBR Green I assay for the rapid and visual detection of Brucella. Folia Microbiol (2023). https://doi.org/10.1007/s12223-023-01115-2
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DOI: https://doi.org/10.1007/s12223-023-01115-2