Porcine epidemic diarrhea, a disease caused by porcine epidemic diarrhea virus (PEDV), results in large economic losses to the global swine industry. To manage this disease effectively, it is essential to detect PEDV early and accurately. We developed a sensitive and accurate droplet digital PCR (ddPCR) assay to detect PEDV. The optimal primer-to-probe concentration and melting temperature were identified as 300:200 nM and 59.2°C, respectively. The specificity of the ddPCR assay was confirmed by negative test results for common swine pathogens. The detection limit for the ddPCR was 0.26 copies/μL, which is a 5.7-fold increase in sensitivity compared to that of real-time PCR (rtPCR). Both ddPCR and rtPCR assays exhibited good linearity, although ddPCR provided higher sensitivity for clinical detection compared to that of rtPCR. Our ddPCR methodology provides a promising tool for evaluating the PEDV viral load when used for clinical testing, particularly for detecting samples with low-copy viral loads.

猪流行性腹泻是一种由猪流行性腹泻病毒（PEDV）引起的疾病，给全球养猪业造成了巨大的经济损失。为了有效地控制这种疾病，必须尽早准确地检测出PEDV。我们开发了灵敏，准确的微滴数字PCR（ddPCR）检测法来检测PEDV。最佳引物与探针的浓度和解链温度分别确定为300：200 nM和59.2°C。ddPCR检测的特异性通过常见猪病原体的阴性测试结果得到证实。ddPCR的检测限为0.26拷贝/μL，与实时PCR（rtPCR）相比，灵敏度提高了5.7倍。ddPCR和rtPCR分析均显示出良好的线性，尽管与rtPCR相比，ddPCR为临床检测提供了更高的灵敏度。