Multiplex RT-LAMP method development for rapid detection of Porcine reproductive and respiratory syndrome virus (PRRSV)

Abstract

    Porcine reproductive and respiratory syndrome (PRRS) is one of the most complex and dangerous diseases in pigs, with a high herd mortality rate. To date, disease diagnosis has been primarily based on clinical signs at the onset of the disease and antibody methods, as RT-PCR molecular diagnosis frequently requires a prolonged period of time and complicated testing techniques. Meanwhile, RT-LAMP inherits all of PCR's advantages in terms of sensitivity and specificity, but with a simpler technique and a shorter implementation time. As a result, the primary goal of this study was to develop a protocol for rapid detection of porcine reproductive and respiratory syndrome virus (PRRSV) using multiplex RT-LAMP. The study's optimized RT-LAMP multiplex procedure detected the PRRSV target sequence with a detection limit of 10^0.5 TCID₅₀/mL; the reaction was carried out at a fixed temperature (65 ℃) for 25 minutes, and the results can be read quickly with the naked eye via the reaction color.