Institute of Animal Husbandry and Veterinary Science, Anhui Academy of Agricultural Sciences, Livestock and Poultry Epidemic Diseases Research Center of Anhui Province, Anhui Province Key Laboratory of Livestock and Poultry Product Safety Engineering, Hefei, China.
Animal Health Supervision Institute, Feixi County Agricultural and Rural Bureau, Hefei, China.
Front Cell Infect Microbiol. 2022 Sep 13;12:976137. doi: 10.3389/fcimb.2022.976137. eCollection 2022.
Porcine epidemic diarrhea virus (PEDV) is an enteric coronavirus that causes acute watery diarrhea and vomiting in unweaned piglets. Infections result in high mortality and serious economic losses to the swine industry. PEDV attenuated vaccine does not completely protect against all mutant wild-type strains, and PEDV infection can periodically occur. A sensitive, accurate, and simple detection method for PEDV is needed to reduce the occurrence of the disease. In this study, the CRISPR/Cas13a system was combined with recombinase aided amplification to develop a rapid diagnostic method to distinguish PEDV wild-type strains from attenuated vaccine strains. The method is based on isothermal detection at 37°C. The results are used for visual readout. The assay had high sensitivity and specificity, with a detection limit of 10 copies/μL for the gene of interest, and no cross-reactivity with other pathogens. The Cas13a detection worked well with clinical samples. This visual, sensitive, and specific nucleic acid detection method based on CRISPR/Cas13a should be a powerful tool for detecting PEDV.
猪流行性腹泻病毒（PEDV）是一种肠道冠状病毒，可导致未断奶仔猪发生急性水样腹泻和呕吐。感染可导致高死亡率和养猪业的严重经济损失。PEDV 减毒疫苗不能完全预防所有突变型野生型毒株，PEDV 感染会周期性发生。需要一种灵敏、准确、简单的 PEDV 检测方法来减少疾病的发生。本研究将 CRISPR/Cas13a 系统与重组酶辅助扩增相结合，开发了一种快速诊断方法，用于区分 PEDV 野生型株和减毒疫苗株。该方法基于 37°C 的等温检测。结果用于直观读出。该测定法具有高灵敏度和特异性，对目的基因的检测限为 10 拷贝/μL，与其他病原体无交叉反应。Cas13a 检测与临床样本配合良好。这种基于 CRISPR/Cas13a 的可视化、灵敏和特异的核酸检测方法应该是检测 PEDV 的有力工具。
