Establishment of Detection Method for Loop-mediated Isothermal Amplification of Porcine Pseudorabies Virus
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Abstract:
To establish a universal rapid detection method for porcine pseudorabies virus (PRV) loop-mediated isothermal amplification, four sets of specific primers were designed based on the conserved segments of the PRV gB gene sequence, and loop primers were introduced based on loop-mediated isothermal amplification. The specific primer PRV-1 was screened, and its specificity and sensitivity were evaluated. Clinical samples and artificial interference samples were tested and compared with real-time quantitative real-time PCR (qPCR). The results showed that the detection method could detect the presence of PRV specifically, and had good specificity. The limit of sensitivity detection is 10 fg/μL. The results of the detection method established in this work were highly correlated with the results of qPCR detection of clinical samples and artificial interference samples. Therefore, the PRV loop-mediated isothermal amplification detection method established in this work is specific, sensitive, and has good stability. The operation is safe, simple and efficient. It is potential to be a rapid diagnosis and grassroots of pseudorabies.
