RAW 数据-一种 crRNACas12a 复合物驱动的 4 例猪腹泻快速视觉检测viruses.zip

猪流行性腹泻病毒（PEDV）、传染性胃肠炎病毒（TGEV）、猪德尔塔冠状病毒（PDCoV）和猪轮状病毒-A（PoRV）G9是主要导致胃肠道疾病的主要猪病原菌，尤其影响泌乳仔猪并造成重大经济损失，尤其是在中国。本研究引入了一种基于CRISPR的新型核酸检测方法，该方法将huLbCas12a的高特异性与环介导的等温扩增（LAMP）技术的灵敏度相结合。该方法的核心是分子量约为 144 kDa 的 crRNA/Cas12a 复合物，通过靶向基因编辑提高诊断准确性。该方法结合荧光报告探针和侧流试纸测定，建立了一个能够同时识别所有四种病毒的视觉检测系统。它能够对病毒基因组进行低至 1 至 10 拷贝/μL 的可视化，而不会出现交叉反应。在对 95 个临床样本的比较测试中，我们的四重 LAMP-CRISPR 检测表明，三种猪冠状病毒与 RT-qPCR 的一致性为100%，PoRV G9 与RT-qPCR的一致性为98.94%。该方法为现场病毒检测提供了强大而可靠的工具，极大地有助于及时预防病毒传播并减轻其对养猪业的影响，证明了其在加强兽医环境中的生物安全和疾病管理方面的关键作用。

Porcine Epidemic Diarrhea Virus (PEDV), Transmissible Gastroenteritis Virus (TGEV), Porcine Delta Coronavirus (PDCoV), and Porcine Rotavirus-A (PoRV) G9 are major swine pathogens that primarily cause gastrointestinal diseases, which particularly afflict lactating piglets and inflict significant economic losses, especially in China. This study reports a novel CRISPR-based nucleic acid detection assay that integrates the high specificity of huLbCas12a with the sensitivity of loop-mediated isothermal amplification (LAMP) technology. The core component of this assay is the crRNA/Cas12a complex with a molecular weight of approximately 144 kDa, which enhances diagnostic accuracy via targeted recognition of viral genomic sequences. By combining fluorescent reporter probes and lateral flow strip assays, this method establishes a visual detection system capable of simultaneously identifying all four target viruses. It allows visualization of viral genomes down to 1–10 copies per μL with no cross-reactivity. In a comparative test of 95 clinical samples, our quadruple LAMP-CRISPR assay showed 100% consistency with RT-qPCR for the three porcine coronaviruses, and 98.94% consistency for PoRV G9 relative to RT-qPCR. This assay provides a robust and reliable tool for on-site viral detection, greatly facilitating timely prevention of viral transmission and mitigation of its impacts on the swine industry, thereby demonstrating its critical role in strengthening biosafety and disease management in veterinary settings.