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ORIGINAL RESEARCH article

Front. Vet. Sci.

Sec. Veterinary Infectious Diseases

Volume 12 - 2025 | doi: 10.3389/fvets.2025.1655079

Establishment and application of a dual chip digital PCR Assay for detection of PDCoV and PEDV

Provisionally accepted
Yue  ZhangYue Zhang1,2Fangting  DongFangting Dong1Yuhang  ZhangYuhang Zhang1Yutong  FengYutong Feng1Jinwang  HuJinwang Hu1Yuhang  LiYuhang Li1Lu  XiaLu Xia1,3Shaopo  ZuShaopo Zu1,3Hao  LuHao Lu4*Zhanyong  WeiZhanyong Wei1,2*
  • 1College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China
  • 2Ministry of Education Key Laboratory for Animal Pathogens and Biosafety, Zhengzhou, China
  • 3Henan Province Key Laboratory for Animal Food Pathogens Surveillance, Zhengzhou, China
  • 4Molecule Biology Laboratory of Zhengzhou Normal University, Zhengzhou, China

The final, formatted version of the article will be published soon.

Coinfection with porcine deltacoronavirus (PDCoV) and porcine epidemic diarrhea virus (PEDV) is a major cause of acute diarrhea in piglets, which poses a significant challenge to the swine industry. Highly sensitive diagnostic tools are required for the early detection and control of these two viruses. We developed a novel chip digital PCR (cdPCR) assay that uses two probes for the simultaneous quantitative detection of both PDCoV and PEDV in clinical samples—including in aerosols with low viral loads. The assay demonstrated exceptional sensitivity, with limits of detection (LoD) of 1.83 ± 0.15 copies/μL for PDCoV and 0.99 ± 0.07 copies/μL for PEDV, high specificity (no cross-reactivity with TGEV, PSV, or PRV), outstanding linearity (R²=0.9972 for PDCoV and R²=0.9969 for PEDV) and reproducibility (intra- and inter-assay CV<6%). Validation across 148 clinical samples indicates that our dual cdPCR is more sensitive than qPCR for detecting both single and mixed infections. Notably, this assay can effectively quantify PDCoV and PEDV in environmental aerosol samples. Our results demonstrate that this dual cdPCR assay offers a highly sensitive, stable, and accurate platform for the simultaneous quantification of both PDCoV and PEDV. It represents a valuable tool for early disease monitoring (particularly in aerosol surveillance and mixed-infection scenarios with low viral loads), thereby supporting the effective prevention of porcine viral diarrhea and the sustainable growth of the swine industry.

Keywords: cdPCR, PDCoV, PEDV, Dual assay, detection

Received: 27 Jun 2025; Accepted: 22 Aug 2025.

Copyright: © 2025 Zhang, Dong, Zhang, Feng, Hu, Li, Xia, Zu, Lu and Wei. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence:
Hao Lu, Molecule Biology Laboratory of Zhengzhou Normal University, Zhengzhou, China
Zhanyong Wei, College of Veterinary Medicine, Henan Agricultural University, Zhengzhou, China

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