Prokaryotic Expression of Novel Duck Reovirus σB Protein and Establishment of an Indirect ELISA Antibody Detection Method
In order to establish an efficient and convenient method for the detection of novel duck reovirus (NDRV) antibodies,an indirect ELISA method for the detection of NDRV antibodies was developed using recombinant σB protein expressed by the E.coli expression system as the encapsulated antigen.The reaction conditions of the assay were optimized,and specificity,sensitivity and reproducibility tests as well as clinical samples were performed.The results showed that the optimal reaction conditions of the established indirect ELISA method were as follows:0.5 μg/mL antigen was encapsulated at 4 ℃ overnight,the serum to be examined was diluted 1∶400 and incubated at 37℃ for 60 min,the color development time was 15 min,and the value of Cut off was 0.432.The method did not cross-react to the positive sera of Newcastle Disease Virus ( NDV ),Duck Circovirus ( DuCV),Novel Goose parvovirus ( NGPV) and Duck Tambusu Virus ( DTMUV),and the sensitivity of detecting the positive sera was 1∶6400,with coefficients of variation of the inter-and intra-batch replicate tests of less than 8％,and the overall compliance rate of this assay with the commercially available ELISA kit for the antibody against NDRV was 96.88％.In this study,an indirect ELISA assay was successfully established,providing an effective and convenient method for NDRV antibody detection.
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