中国生物工程杂志

Abstract：Based on the published nucleotide sequences of Influenza A virus, Porcine reproductive and respiratory syndrome virus, Porcine cirocovirus-2 and Porcine pseudorabies virus, 4 pairs of primers were designed. And conserved DNA (or cDNA) fragments of the 4 viruses obtained by using PCR (or RT-PCR), were ligated with pMD 18-T Simple Vector to construct recombinant plasmid for DNA probe preparation. The recovered DNA probes were spotted on located sites on nitrocellulose filter to prepare low density DNAarray. During the process of multiplex PCR, samples were labeled by Biotin-11-dUTP. Then the labeled PCR products were hybridized with DNAarrays, and the hybridization signals were scanned and analyzed by scanning device and software. The results showed that this DNAarray could identify and distinguish the 4 viruses in samples synchronously and was more sensitive than PCR method. This method can be use for lab diagnosis, large-scale epidemiology investigation and quarantine of animal and animal products.