Journal Article

Development and Application of a TaqMan RT-qPCR for the Detection of Foot-and-Mouth Disease Virus in Pigs

Sun, Yajuan, Zhao, Hongri, Yin, Rui, Wang, Meiqi, Xiao, Xingyu, Dong, Changying, Zhang, Yongzhe, Cao, Qianyue, Yang, Yanran, Jin, Hui

2024

Overview

The global livestock industry is facing a serious threat from a widespread foot-and-mouth disease virus (FMDV) epidemic. The timely detection of FMDV can significantly mitigate its harmful effects. This study aimed to establish and evaluate a TaqMan fluorescence quantitative PCR assay to assess its sensitivity, specificity, reproducibility, and stability. The standard curve equation range is 6.43 × 109–6.43 × 101 copies/µL, with an R2 value of 0.996 and a standard curve equation of y = −3.586x + 36.245. The method successfully detected 64.3 copies/µL of the target gene for FMDV and exhibited high specificity for FMDV. Repeatability tests demonstrated low coefficients of variation within and between groups (<2%), indicating good reproducibility. The clinical samples analyzed using this method showed results consistent with those of the SYBR Green I RT-qPCR assay, confirming the reliability of the method. Overall, the developed test method displayed high sensitivity, specificity, reproducibility, and stability, making it suitable for the rapid diagnosis of foot-and-mouth disease in clinical settings.

• Publisher: MDPI AG,MDPI
• Subject: Annealing / Antigens / Bioengineering / Communicable diseases / detection methods / Disease transmission / Enzymes / Foot & mouth disease / Foot-and-mouth disease / foot-and-mouth disease virus / Genomes / Hogs / Infectious diseases / Plasmids / Swine / system optimization / TaqMan real-time RT-qPCR / Vaccines / Virulence / Viruses