Development and Application of Colloidal Gold Test Strip Against Porcine Rotavirus
[Objective]Two strains of monoclonal antibodies of Porcine rotavirus(PoRV)were used to establish PoRV colloidal gold test strip for rapid detection of PoRV.[Method]The 40 nm colloidal gold particles were used to label the monoclonal antibody 4G5.The monoclonal antibody 5G4 and sheep anti-mouse IgG were coated on the surface of nitrocellulose membrane as the detection line and the control line,respectively.The colloidal gold test strip was established by optimizing the optimal pH of colloidal gold solution,the optimal usage amount of the labeled antibody and the detection antibody.The sensitivity,specificity,repeatability and stability of the strip were assessed.The coincidence rate of the strip and RT-PCR was calculated by detecting 50 pig fecal samples,and then the strip was put to clinical use by detecting 1 249 clinical samples.[Result]The optimal parameters of the test strip were as follows:The optimal pH of colloidal gold solution was 8.0,the optimal usage amounts of labeled antibody 4G5 and detection antibody 5G4 were 14.4 μg/mL and 1 mg/mL,respectively.The strip could detect a 1∶2 000 dilution of PoRV OSU strain(genotype G5),the sensitivity was 104.5 TCID50/mL,and the detection results of PoRV G9,G3 and G4 strains were all positive.And no cross reaction occurred with Transmissible gastroenteritis virus(TGEV),Porcine epidemic diarrhea virus(PEDV),Porcine deltacoronavirus(PDCoV),Porcine pseudorabies virus(PRV)and Porcine circovirus type 2(PCV2).The assay results showed that the strip had good repeatability in and between batches and stability.50 pig fecal samples were detected by the strip and RT-PCR,the results showed that the positive coincidence rate was 97.5％(39/40),the negative coincidence rate was 100％(10/10),and the overall coincidence rate was 98％(49/50).1 249 clinical samples which were collected from pig farms in Henan and Shanxi provinces in 2022 were detected by the test strip,and the positive rate of PoRV was 5.2％.[Conclusion]The colloidal gold strip developed in this study had good sensitivity,specificity and repeatability,and high coincidence rate with RT-PCR method,and could be used for PoRV rapid detection in industrialized pig farm.
Porcine rotavirus(PoRV)colloidal gold test striprapid detection
万方数据
维普
