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Updated: May 21, 2026
Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Assay for the Specific and Rapid Detection of Tilapia Lake Virus
Published on: May 18, 2020
A direct and rapid RT-LAMP method for Point-of-Care test of porcine deltacoronavirus
Fu Zhu1, Xueqing Zong2, Xiaoqing Xie2
1National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China; Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, Guangzhou 510640, China.
Abstract:
Porcine deltacoronavirus (PDCoV) is a significant enteric pathogen in swine, necessitating rapid and accessible diagnostic tools for effective control. In this study, we developed a streamlined, extraction-free Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) assay integrated with a TaqMan probe for on-site PDCoV detection. By targeting the conserved N gene and modifying the loop primer (LB) as a fluorescence probe, a highly specific reaction system was established. The assay demonstrated high sensitivity, with a limit of detection (LOD) of 9.3 copies/μL for cDNA and 5.9 copies/μL for RNA within 40 min, while showing no cross-reactivity with common swine viruses. Reproducibility assays yielded a coefficient of variation (CV) of < 2.5%, indicating excellent stability. Furthermore, the method was successfully integrated with nucleic acid extraction-free technology for fecal sample analysis. In clinical validation using 75 samples, the assay showed 100% concordance with column-based RT-LAMP and standard RT-qPCR methods. This extraction-free RT-LAMP-TaqMan assay offers a rapid, reliable, and cost-effective Point-of-Care Testing (POCT) solution for PDCoV surveillance in swine herds.
