Shinuo Cao
1
Dongxue Ma
2
Jun Xie
1*
Zhi Wu
1
Shengwei Ji
2
Zhou Mo
1*The final, formatted version of the article will be published soon.
ORIGINAL RESEARCH article
Front. Cell. Infect. Microbiol.
Sec. Veterinary and Zoonotic Infection
Volume 14 - 2024 |
doi: 10.3389/fcimb.2024.1474825
Point-of-care testing Diagnosis of African Swine Fever Virus by Targeting Multiple Genes with Enzymatic Recombinase Amplification and CRISPR/Cas12a System
Provisionally accepted- 1 Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-animal Husbandry Vocational College, Taizhou, China
- 2 Department of Veterinary Medicine, College of Agriculture, Yanbian University, Yanji, Jilin Province, China
- 3 MOA Key Laboratory of Animal Bacteriology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, China, Nanjing, China
African swine fever virus (ASFV) infection is causing devastating outbreaks globally; pig farming has suffered severe economic losses due to the ASFV. Currently, strict biosecurity control measures can mitigate the incidence of ASF. Rapid, cost-effective, and sensitive detection of ASFV can significantly reduce disease transmission and mortality. CRISPR/Cas-associated proteins can detect polymorphisms with high specificity and sensitivity, making them ideal for detecting pathogens. In this study, based on CRISPR/Cas12a integrated with enzymatic recombinase amplification (ERA) technology, a CRISPR/Cas12a detection system capable of identifying ASFV E183L, K205R, and C962R gene sequences has been developed. The ERA-CRISPR/Cas12a detection system detected ASFV precisely without cross-reactivity with other porcine pathogen templates and with a sensitivity detection limit of 10 copies per reaction; it takes 60 minutes to complete the detection process. In combination with this integrated ERA pre-amplification and Cas12a/crRNA cutting assay, it provides a rapid, straightforward, sensitive, and specific method for ASFV detection in the field.
Keywords: African Swine Fever Virus, Point-of-care testing, Enzymatic recombinase amplification, CRISPR/Cas12a, diagnosis
Received: 02 Aug 2024; Accepted: 04 Nov 2024.
Copyright: © 2024 Cao, Ma, Xie, Wu, Yan, Ji, Mo and Zhu. 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:
Jun Xie, Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-animal Husbandry Vocational College, Taizhou, China
Haoyu Yan, MOA Key Laboratory of Animal Bacteriology, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, China, Nanjing, China
Zhou Mo, Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-animal Husbandry Vocational College, Taizhou, China
Shanyuan Zhu, Engineering Technology Research Center for Modern Animal Science and Novel Veterinary Pharmaceutic Development, Jiangsu Agri-animal Husbandry Vocational College, Taizhou, China
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