Development and application of N-protein blocking ELISA for detecting porcine epidemic diarrhea virus antibodies
Acta Veterinaria et Zootechnica Sinica
; 53(4):1173-1181, 2022.
Article
in Chinese
| CAB Abstracts | ID: covidwho-1975364
ABSTRACT
The purpose of this study is to establish a blocking ELISA antibodies detection method for porcine epidemic diarrhea virus (PEDV). The purified N protein was used as the coating antigen, and the ELISA reaction conditions were optimized by the chess rboard titration. A blocking ELISA method for detecting PEDV antibodies was established, and its specificity, sensitivity and repeatability tests were carried out. One hundred and forty clinical serum samples were tested, and the results were compared with commercially IDvet PEDV indirect ELISA antibodies detection kit. The results showed that the best antigen coating concentration was 625 ng.mL-1, and the best dilution ratio of serum was 11;The best dilution of the HRP-conjugated antibody working solution was 15 000;There was no cross-reaction with healthy pig serum and the positive sera of common pig disease pathogens, such as classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), and transmissible gastroenteritis virus (TGEV). The sensitivity of PEDV positive serum was 116, which was equivalent to that of IDvet ELISA kit (titer 132). The coefficient of variation of within-run and between-run repeatability test is less than 10%, so it showed that the blocking ELISA established in this study had good repeatability and stability;the kappa value of detected 140 clinical porcine serum using this method was 0.87 when compared with IDvet ELISA. The above results indicated that the established blocking ELISA method for detecting PEDV antibodies in this study could be applied to the prevention and control of PEDV, epidemiological investigation and the monitoring of antibody levels after vaccine immunization.
Meat Producing Animals [LL120]; Prion, Viral, Bacterial and Fungal Pathogens of Animals [LL821]; Diagnosis of Animal Diseases [LL886]; Techniques and Methodology [ZZ900]; antibodies; antibody testing; diagnosis; diagnostic techniques; ELISA; immunodiagnosis; porcine epidemic diarrhoea; viral antigens; pigs; Porcine epidemic diarrhea virus; China; APEC countries; East Asia; Asia; high Human Development Index countries; upper-middle income countries; Sus scrofa; Sus; Suidae; Suiformes; Artiodactyla; mammals; vertebrates; Chordata; animals; eukaryotes; Alphacoronavirus; Coronavirinae; Coronaviridae; Nidovirales; positive-sense ssRNA Viruses; ssRNA Viruses; RNA Viruses; viruses; antibody detection; antibody tests; People's Republic of China; enzyme linked immunosorbent assay; serological diagnosis; swine; hogs; Porcine epidemic diarrhoea virus
Full text:
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Collection:
Databases of international organizations
Database:
CAB Abstracts
Language:
Chinese
Journal:
Acta Veterinaria et Zootechnica Sinica
Year:
2022
Document Type:
Article
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