ORIGINAL RESEARCH article
Front. Vet. Sci.
Sec. Veterinary Infectious Diseases
Establishment and field validation of a rapid on-site recombinase polymerase amplification– lateral flow assay for BRSV and BVDV
Provisionally accepted
LILI CAO1*
Zhiteng Zhao2
Yanbing Guo1Shaoxiong Liu2Liangyu Hao1Xingzhong Sun1Yu Xiang1Nan Wang1Xiangyu Meng3Hongbo Sun4Shuai Yue5
Pengtao Gong2- 1Jilin Academy of Animal Husbandry and Veterinary Medicine, changchun, China
- 2State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases and College of Veterinary Medicine, Jilin University, Changchun, China
- 3Animal Husbandry Development Service Center, Tongyu County, Baicheng, China
- 4Animal Disease Prevention and Control Center, Tongyu County, Baicheng, China
- 5Animal Disease Prevention and Control Center, Xing‘an League, China
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Bovine respiratory disease (BRD) is the costliest bovine syndrome worldwide, inflicting annual losses of over one billion USD in North America alone. Transport stress, overcrowding and viral–bacterial synergy can drive mortality to 70 %, yet laboratory-based diagnostics delay decisive treatment. We therefore developed pen-side real-time enzymatic recombinase amplification lateral-flow dipsticks (RT-ERA-LFD) assays targeting the two principal viral agents, bovine respiratory syncytial virus (BRSV) and bovine viral diarrhoea virus (BVDV), which enables their separate detection in a single tube. The BRSV nucleoprotein gene and BVDV 5'-UTR were cloned and in-vitro transcribed into quantified RNA standards to calibrate an enzymatic recombinase amplification (ERA) coupled with lateral-flow dipsticks (LFD). After primer/probe optimisation (BRSV-ERA-F1/R4/P2; BVDV-ERA-F1/R4/P1), the 40 °C, 20-min reactions detected as few as ten template copies, showed 100 % specificity against related bovine pathogens and matched real-time PCR results in 46 archived respiratory samples. In a field survey of nasal swabs from cattle farms in Jilin Province, China, BRSV was detected in 10.87 % and BVDV in 8.70 % of specimens, with results identical to qPCR obtained within 30 min without instrumentation. By delivering actionable infection status at the chute, the platform enables mass screening and timely intervention, effectively mitigating BRD's global economic impact.
Keywords: BRSV, BVDV, Enzymatic recombinase amplification, ERA-LFD, lateral-flow dipsticks
Received: 26 Nov 2025; Accepted: 14 Feb 2026.
Copyright: © 2026 CAO, Zhao, Guo, Liu, Hao, Sun, Xiang, Wang, Meng, Sun, Yue and Gong. 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: LILI CAO
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