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. 2017 Nov:249:76-78.
doi: 10.1016/j.jviromet.2017.08.020. Epub 2017 Aug 30.

Development and application of a recombinant M protein-based indirect ELISA for the detection of porcine deltacoronavirus IgG antibodies

Shang-Xing Luo  1 , Jing-Hui Fan  1 , Tanja Opriessnig  2 , Jing-Mei Di  1 , Bao-Jing Liu  1 , Yu-Zhu Zuo  3
Affiliations

Development and application of a recombinant M protein-based indirect ELISA for the detection of porcine deltacoronavirus IgG antibodies

Shang-Xing Luo et al. J Virol Methods. 2017 Nov.

Abstract

Porcine deltacoronavirus (PDCoV) is a recently identified coronavirus in the genus Deltacoronavirus that can cause enteric disease including diarrhea, vomiting, dehydration and mortality in neonatal piglets. Serological assays to detect anti-PDCoV antibodies are presently limited to certain laboratories and geographic regions. In this study, a recombinant M protein-based indirect enzyme-linked immunosorbent assay (PDCoV-rM ELISA) was developed and utilized to determine the prevalence of anti-PDCoV IgG in Hebei province. The PDCoV-rM ELISA showed no cross-reaction with antisera against transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), porcine rotavirus (PRV), porcine circovirus 2 (PCV2), classical swine fever virus (CSFV) or porcine reproductive and respiratory syndrome virus (PRRSV). The diagnostic sensitivity was 90.6% and the diagnostic specificity was 93.3%. A total of 871 serum samples collected in Hebei from January 2015 to October 2016 were checked for presence of antibodies against PDCoV using the novel PDCoV-rM ELISA. Anti-PDCoV IgG antibodies were detected in 11% (96/871) of the samples and in 25% (10/40) of the investigated farms. The data suggest that PDCoV has a low seroprevalence in pig population in Hebei province, China.

Keywords: ELISA; M protein; Porcine deltacoronavirus; Serum epidemiology.

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Figures

Fig. 1
Fig. 1
Expression and identification of PDCoV-rM. A: Prokaryotic expression of the PDCoV-rM: Lane 1,Protein marker (14–100 kDa); Lane 2, IPTG-induced recombinant bacteria with the pET-32a vector for 4 h; Lane 3, Uninduced recombinant bacteria with PDCoV-rM; Lane 4, IPTG-induced recombinant bacteria with PDCoV-rM for 1 h; Lane 5, IPTG-induced recombinant bacteria with PDCoV-rM for 2 h; Lane 6, IPTG-induced recombinant bacteria with the PDCoV-rM vector for 3 h. Lane 7, IPTG-induced recombinant bacteria with the PDCoV-rM vector for 4 h. B: Western blot of the recombinant PDCoV-rM. Lane 1, IPTG-induced recombinant bacteria with the pET-32a vector for 4 h; Lane 2, purified PDCoV-rM; Lane 3, Protein marker (14–100 kDa).
Fig. 2
Fig. 2
Distribution of anti-PDCoV IgG antibodies in serum samples obtained from farms with known PDCoV infection. Serum samples were classified as negative or positive based on viral RNA detection on fecal samples of the pigs. Data presented as ELISA OD values ± SEM. The assay cut-off (OD value of 0.35) is indicated by the dashed line.
See this image and copyright information in PMC

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