Method For Rapid Detection Of PRV GE Protein Antibody

Porcine pseudorabies(PR),a disease caused by pseudorabies virus(PRV),breaks out in a variety of animals.Swine are the main source and natural host of PRV.At the end of 2011,PR outbroke among Bartha K-61 vaccinated swine,causing huge economic loss.Studies show that PRV variants have emerged.The DIVA(differentiating infected from vaccinated animals)strategy is effective for preventing and controlling of PR.To achieve the goal of eradicating pseudorabies,rapid and effective PRV gE protein detection method is necessary.The gE protein of the PRV variant HNBA strain was obtained by the E.coil expression system.With our purified gE recombinant protein,a series of methods for detecting gE antibodies were established.First,an indrect ELISA(i-ELISA)for the gE protein antibody detection was developed.Compared with blocking gE-ELISA commercial kits,the diagnostic sensitivity,diagnostic specificity,and agreement of i-ELISA were 76.92%,94.12%,and 91.36%,respectively.Based on the above work,a colloidal gold immunochromatographic assay(GICA)for detecting antibodies of PRV gE protein was established.The gE protein and the swine IgG were used as the detection line(T-line)and the quality control line(C-line)respectively.And the coating concentration of both gE protein and the swine IgG is 2 mg/mL.The results of sensitivity test showed that the optimal dilution of serum is 1:640,at which T-line and C-line were clear.However,the GICA shows a poor specificity.Then a rapid magnetic-bead-based method for detection of gE antibodies was estabilished.This method can detect the antibodies of the high titer positive serum diluted 51 200,and the linear range is from 50 to 25 600 diluted serum,and the sensitivity is higher than i-ELISA method.In summary,this work developed three methods to detect PRV gE protein antibodies: i-ELISA,GICA,and magnetic-bead-based method.Due to its rapidity,simplicity,sensitivity,and specificity,magnetic-bead-based method is expected to develop a new commercial kit for rapid detection of PRV gE protein antibodies.