可视化RPA-LFD技术快速检测猪链球菌

doi:10.11843/j.issn.0366-6964.2022.02.020

Abstract

Abstract: Here, we describe the development of a visual and easy-to-operate rapid detection technology for Streptococcus suis. Specific primers were designed for the sequence of the species-specific gene glutamate dehydrogenase (gdh) of S. suis. The recombinant polymerase amplification combined with a lateral flow dipstick (RPA-LFD) detection technology was established, which can detect all serotypes of S. suis simultaneously. The reaction temperature and time were optimized, and its specificity and sensitivity were evaluated. At the same time, 45 samples of suspected S. suis infection were detected by this method. The results showed that the detection sensitivity of S. suis RPA-LFD method can reach 100 copies·μL^-1, which is superior to the conventional PCR method and does not cross-react with other common pathogens. The amplification reaction can be carried out efficiently in a wide temperature range (30-45℃), and the detection can be completed in 20-30 min. Using 45 clinical samples of suspected S. suis infection to evaluate the detection system, the detection rate of RPA-LFD is higher than that of bacterial isolation and conventional PCR method, indicating that it has strong practicability. The RPA-LFD detection method of S. suis established in this study has the characteristics of strong specificity, high sensitivity, easy operation. At the same time, it does not rely on instruments and professional operators, and is suitable for field and on-site detection.

Key words: Streptococcus suis, recombinase polymerase amplification, lateral flow dipstick, rapid detection

CLC Number:

S852.61

ZHANG Shanshan, HE Bin, LI Shuguang, LIU Mingcheng, JIANG Jinqing, HU Jianhe, LEI Liancheng, SHEN Zhiqiang, XIA Xiaojing. Rapid Detection of Streptococcus suis with Visual RPA-LFD Technology[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 538-547.

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Rapid Detection of Streptococcus suis with Visual RPA-LFD Technology

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