Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610064, China.
Appl Microbiol Biotechnol. 2023 Sep;107(18):5739-5747. doi: 10.1007/s00253-023-12690-2. Epub 2023 Jul 21.
Porcine deltacoronavirus (PDCoV) is an enteropathogen that causes diarrhea in piglets and may undergo cross-species transmission. The prevention and control of PDCoV are complicated, and a sensitive, specific, and accessible method of diagnosis would be advantageous. Whereas qPCR is a standard approach for detecting PDCoV, it is not effectively sensitive. In the present study, we report such a strategy using an RT-PCR-based RspCas13d detection system and its efficacy in clinical sample diagnosis. The detection limit of this method was 4 copies/μL and no cross-reaction with other viruses such as the porcine epidemic diarrhea virus, classical swine fever virus, pseudorabies virus, porcine reproductive and respiratory syndrome virus, transmissible gastroenteritis virus and porcine rotavirus. The method was also effective in clinical samples. In summary, we demonstrate that RT-PCR-based RspCas13d detection system is an extremely sensitive and specific nucleic acid-based approach for detecting PDCoV. KEY POINTS: • RspCas13d can be used as a candidate molecular diagnostic tool to diagnose viral genomes. • A novel method is proposed using an RT-PCR-based RspCas13d detection system and its effectiveness in the detection of PDCoV. • The RT-PCR-based RspCas13d detection system has excellent sensitivity and specificity.
猪德尔塔冠状病毒 (PDCoV) 是一种引起仔猪腹泻的病原体，可能发生跨种传播。PDCoV 的预防和控制较为复杂，一种敏感、特异且易于获得的诊断方法将具有优势。虽然 qPCR 是检测 PDCoV 的标准方法，但它的灵敏度并不高。在本研究中，我们报告了一种使用基于 RT-PCR 的 RspCas13d 检测系统的策略及其在临床样本诊断中的效果。该方法的检测限为 4 拷贝/μL，与其他病毒如猪流行性腹泻病毒、猪瘟病毒、伪狂犬病病毒、猪繁殖与呼吸综合征病毒、传染性胃肠炎病毒和猪轮状病毒无交叉反应。该方法在临床样本中也有效。总之，我们证明了基于 RT-PCR 的 RspCas13d 检测系统是一种非常敏感和特异的基于核酸的 PDCoV 检测方法。
RspCas13d 可作为一种候选分子诊断工具，用于诊断病毒基因组。
提出了一种使用基于 RT-PCR 的 RspCas13d 检测系统的新方法，及其在 PDCoV 检测中的有效性。
基于 RT-PCR 的 RspCas13d 检测系统具有出色的灵敏度和特异性。
