243 results about "Porcine epidemic diarrhea virus" patented technology

The invention discloses a recombinant porcine epidemic diarrhea virus (PEDV) lactococcus lactis and its construction method and use. The construction method comprises the following steps of carrying out RT-PCR amplification to obtain a major protective antigen COE gene sequence of PEDV, carrying out clone sequencing, designing expression primers according to the sequencing result and expression vector characteristics, carrying out amplification of a PEDV COE coding sequence, connecting the PEDV COE coding sequence to a lactococcus lactis expression vector pNZ8149, and carrying out electrotransformation of the lactococcus lactis expression vector pNZ8149 into a lactococcus lactis NZ3900 cell to obtain the recombinant PEDV lactococcus lactis. The recombinant PEDV lactococcus lactis is induced by 1ng/mL of Nisin and through SDS-PAGE and Western blot analysis, a main s protein of the PEDV is expressed. An expressed recombinant PEDV lactococcus lactis vector oral vaccine can irritate production of humoral immunity and certain cellular immunity of mice. The recombinant PEDV lactococcus lactis has wide application prospects in drugs such as a PEDV vaccine.

The invention discloses a DPO (Dual Priming Oligonucleotide) primer group for porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus and porcine rotavirus detection and application of the DPO primer group. Aiming to a TGEV-N gene, a PEDV-N gene and a PRoV-VP7 gene, a pair of DPO primers respectively, and a multiplex DPO real-time RT-PCR detection method of a porcine epidemicdiarrhea virus, a porcine transmissible gastroenteritis virus and a porcine rotavirus is established. Results show that a detection limit of the method is 5.3*100 copies/microlitre, target gene fragments can be efficiently amplified within an annealing temperature range from 40 DEG C to 65 DEG C, and shown that the method is wide in annealing temperature range; and meanwhile, the DPO primers are strong in specificity, and no non-specific amplification is generated in the PCR process. The multiplex DPO real-time RT-PCR detection method provided by the invention is simple to design, strong in specificity and high in sensitivity and provides a new technological means to rapid and accurate detection of three porcine viral diarrhea infective pathogens of TGEV, PEDV and PRoV.