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

Simple Summary: Foot-and-mouth disease (FMD), caused by the foot-and-mouth disease virus (FMDV), is a prevalent infectious disease in swine. Timely detection and isolation of pigs infected with this virus are among the most effective strategies to prevent the disease's spread and mitigate potential economic damage. We have established a set of RT-qPCR detection methods for FMDV, which offer significant advantages. Notably, this detection method can effectively identify foot-and-mouth disease virus even in the presence of both exogenous and endogenous interfering substances. Furthermore, it exhibits high sensitivity, strong specificity, and good repeatability, enabling 100% detection of FMDV. 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. [ABSTRACT FROM AUTHOR]