Abstract: The aim of this study is to establish a double RT-PCR assay for simultaneously detecting porcine epidemic diarrhea virus（PEDV）and porcine delta corona virus（PDC_OV）. Two pairs of specific primers were designed according to the sequences of PEDV and PDC_OV genes published in GenBank. Firstly,the RT-PCR reaction technique was used to optimize the reaction conditions through the single amplification of the target genes of PEDV and PDC_OV viruses. Then,the establishment of double RT-PCR method was confirmed by specificity test,sensitivity test and clinical sample detection. Results showed as：The size of the amplified target gene PEDV-M was 750 bp,and 372 bp for PDC_OV-N fragment;PEDV and PDC_OV genes were detected simultaneously,but 3 viruses of PRV,CSFV and PPV were not detected. The minimum concentration limit of mixed nucleic acid concentration for optimizing and amplifying PEDV-PDC_OV was 100 pg/μL,thus it can be used to identify the clinical suspected virus infection. The results showed that a double RT-PCR detection method for PEDV-PDC_OV virus is established successfully,this method is highly sensitive and specific and a rapid and effective method for the clinical detection of PEDV-PDC_OV single virus infection or multi-virus mixed infection in pigs.

Key words: PEDV, PDC_OV, RT-PCR, clinical diagnosis