Pseudorabies viruses (PRV) pose a major threat to the global pig industry and public health. Rapid, intuitive, affordable, and accurate diagnostic testing is critical for controlling and eradicating infectious diseases. In this study, a portable detection platform based on RPA-CRISPR/EsCas13d was developed. The platform exhibits high sensitivity (1 copy/μL), good specificity, and no cross-reactivity with common pathogens. The platform uses rapid preamplification technology to provide visualization results (lateral flow assays or visual fluorescence) within 1 h. Fifty pig samples (including tissues, oral fluids, and serum) were tested using this platform and real-time quantitative polymerase chain reaction (qPCR), showing 34.0 % (17 of 50) PRV positivity with the portable CRISPR/EsCas13d dual-readout platform, consistent with the qPCR results. These results highlight the stability, sensitivity, efficiency, and low equipment requirements of the portable platform. Additionally, a novel point-of-care test is being developed for clinical use in remote rural and resource-limited areas, which could be a prospective measure for monitoring the progression of pseudorabies and other infectious diseases worldwide.

中文翻译：

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使用基于 RPA-CRISPR/EsCas13d 的双读出便携式平台快速、灵敏、直观地检测伪狂犬病病毒


伪狂犬病病毒（PRV）对全球养猪业和公共卫生构成重大威胁。快速、直观、经济且准确的诊断测试对于控制和根除传染病至关重要。本研究开发了基于RPA-CRISPR/EsCas13d的便携式检测平台。该平台具有高灵敏度（1 copy/μL）、良好的特异性，与常见病原体无交叉反应。该平台采用快速预放大技术，可在 1 小时内提供可视化结果（侧流分析或视觉荧光）。使用该平台和实时定量聚合酶链式反应 (qPCR) 对 50 份猪样本（包括组织、口腔液和血清）进行了测试，便携式 CRISPR/EsCas13d 双读出平台显示 PRV 阳性率为 34.0%（50 份中的 17 份） ，与qPCR结果一致。这些结果凸显了便携式平台的稳定性、灵敏度、效率和低设备要求。此外，正在开发一种新颖的护理点测试，用于偏远农村和资源有限地区的临床使用，这可能是监测全球伪狂犬病和其他传染病进展的前瞻性措施。