猪流行性腹泻病毒(PEDV)是一种肠道冠状病毒,可引起断奶仔猪急性水样腹泻和呕吐。感染导致高死亡率和对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的野生型菌株,PEDV感染可定期发生。一个敏感的,准确,需要简单的PEDV检测方法来减少疾病的发生。在这项研究中,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,以区分PEDV野生型株和减毒疫苗株.该方法基于37°C的等温检测。结果用于视觉读出。该方法具有较高的灵敏度和特异性,目的基因的检测限为101拷贝/μL,与其他病原体没有交叉反应。Cas13a检测在临床样品中效果良好。这个视觉,敏感,基于CRISPR/Cas13a的特异性核酸检测方法应该是检测PEDV的有力工具。
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 101 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.