Establishment and preliminary application of an ELISA method for detection of porcine epidemic diarrhea virus IgA antibody
To identify the IgA antibody against porcine epidemic diarrhea virus(PEDV),an ELISA technique was developed and refined using PEDV recombinant S protein as the coating antigen and horseradish peroxidase-conjugated murine anti-porcine IgA monoclonal antibody as the identifying an-tibody,and its specificity,sensitivity,reproducibility,and performance of clinical samples were evaluated.The findings indicated that the optimal antigen encapsulation mass concentration was 2 μg/mL,the best enzyme-labeled antibody working concentration was 1∶25 000,and the most effective serum di-lution was 1∶50.There was no cross-reactivity with positive serums of porcine pseudorabies virus,African swine fever virus,classical swine fever virus,Senecavirus A and foot-and-mouth disease virus.The sensitivity was determined to be 1∶800.The coefficients of variation were less than 10％within and among different batches,and the consistency rate with IDEXX commercial kit was very high in testing result.These results prove that the detection method for PEDV IgA antibody developed in this research has good specificity,sensitivity and reproducibility,making it suitable for clinical diagnosis,di-sease prevention,and control of porcine epidemic diarrhea,as well as for epidemiological studies and evaluation of vaccination efficacy.
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