The invention discloses a porcine
delta-
coronavirus double-
antibody sandwich ELISA (
enzyme-linked immuno
sorbent assay) detection kit based on an S
protein monoclonal antibody as well as a use method and application of the porcine
delta-
coronavirus double-
antibody sandwich ELISA detection kit. A PDCoV S
protein is expressed by adopting an ExpiCHO expression
system, then the purified S
protein is used as an
immunogen to immunize a mouse, and the
monoclonal antibody is obtained. And then, respectively using the
monoclonal antibody 3D7 and the MAb 9G4 labeled by the HRP as capture and detection antibodies, and establishing the double-antibody sandwich
ELISA kit. The kit has good sensitivity, the
detection limit of purified S protein is 0.12 ng / mL, and the
detection limit of PDCoV is 1.96 * 10 < 3 > copies / [mu] L. Specificity analysis shows that the kit has no
cross reactivity with other porcine intestinal coronaviruses PEDV, TGEV and PoRV with clinical symptoms similar to PDCoV, the consistency with an RT-PCR (reverse transcription-
polymerase chain reaction) method reaches 91.03%, and the Kappa value is 0.814, so that the reliability of the kit for clinical sample detection is shown. In addition, the kit can also quantitatively evaluate the content of S protein in PDCoV inactivated vaccines and
subunit vaccines, and provides a powerful tool for evaluating the vaccines.