ORIGINAL RESEARCH article
Front. Cell. Infect. Microbiol.
Sec. Veterinary and Zoonotic Infection
Volume 15 - 2025 | doi: 10.3389/fcimb.2025.1526516
This article is part of the Research TopicBiomarker Identification and Novel Therapies in Equine MedicineView all 3 articles
Integrated CRISPR-Cas12a and RAA One-Pot Visual Strategy for the Rapid Identification of Streptococcus equi Subspecies equi
Provisionally accepted
- 1State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China
- 2State Key Laboratory for Animal Disease Control and Prevention,, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China
- 3College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu Province, China
- 4Country Institute of Western Agriculture, Chinese Academy of Agricultural Sciences (CAAS), Beijing, Beijing Municipality, China
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Strangles, a highly contagious disease caused by Streptococcus equi subspecies equi (S.equi), significantly impacts horse populations worldwide, with Iceland as the only exception. This disease poses serious threats to equine health and results in considerable economic losses. Consequently, the accurate, sensitive, and rapid detection of S.equi from clinical samples is essential for early warning and effective disease management. This study introduces a novel detection method that integrates recombinase-aided amplification (RAA) with CRISPR/Cas12a technologies. We specifically designed RAA primers and CRISPR RNA to target the eqbE gene of S.equi, and we have carefully optimized the reaction systems for this purpose. The newly established visual diagnostic method has shown to be highly effective, demonstrating 97.14 % specificity and 100 % sensitivity, with the capability to detect as few as 5.6×10 0 copies of the target. This is the first study to propose the combined application of RAA and CRISPR/Cas12a for the on-site rapid detection of S.equi. This is the first study to propose the combined application of RAA and CRISPR/Cas12a for the on-site rapid detection of S.equi, which enables visual point-of-care diagnosis of Strangles.
Keywords: Strangles, Streptococcus equi subspecies equi, visual detection, RAA, CRISPR/Cas12a
Received: 06 Apr 2025; Accepted: 23 Jul 2025.
Copyright: © 2025 Zu, Sun, Li, Guo, Wang, Guo and Wang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Wei Guo, State Key Laboratory for Animal Disease Control and Prevention,, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China
Xiaojun Wang, State Key Laboratory for Animal Disease Control and Prevention,, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, China
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