1.Establishment and application of a colloidal gold immunochromatographic test strip assay for detection of caprine enterovirus
Xuebo ZHENG ; Xuyuan CUI ; Naitian YAN ; Qianying WANG ; Hongwei LI ; Lihui CHEN ; Wenjie GONG ; Xinping WANG
Chinese Journal of Veterinary Science 2025;45(4):662-667
To establish an immunochromatographic method for rapid detection of caprine enterovir-us(CEV),the monoclonal antibody against CEV VP1 protein was used as gold-labeled monoclonal antibodies,and the purified rabbit-derived polyclonal antibody of CEV-VP1 and sheep anti-mouse IgG were used as the detection line and quality control line,respectively.The colloidal gold immu-nochromatographic test strips for CEV were prepared according to the principle of double antibody sandwich,evaluated,and applied for clinical specimen detection.The results showed that the meth-od specifically recognized CEV without cross-reaction with bovine enterovirus and bovine viral di-arrhea virus.The minimum detection limit of the method was 102.49 TCID50/mL and had good re-producibility.The prepared test strips had a shelf life of three months kept at 4 ℃.Detection of clin-ical samples using the immunochromatographic test strips showed 100%coincidence rate with RT-PCR method.In conclusion,the colloidal gold immunochromatographic test strips for detection of the emerging CEV with good specificity,sensitivity and repeatability,which provides a new techni-cal means easily used for the rapid detection/diagnosis and epidemiological investigation on CEV infection.
2.Establishment and application of a colloidal gold immunochromatographic test strip assay for detection of caprine enterovirus
Xuebo ZHENG ; Xuyuan CUI ; Naitian YAN ; Qianying WANG ; Hongwei LI ; Lihui CHEN ; Wenjie GONG ; Xinping WANG
Chinese Journal of Veterinary Science 2025;45(4):662-667
To establish an immunochromatographic method for rapid detection of caprine enterovir-us(CEV),the monoclonal antibody against CEV VP1 protein was used as gold-labeled monoclonal antibodies,and the purified rabbit-derived polyclonal antibody of CEV-VP1 and sheep anti-mouse IgG were used as the detection line and quality control line,respectively.The colloidal gold immu-nochromatographic test strips for CEV were prepared according to the principle of double antibody sandwich,evaluated,and applied for clinical specimen detection.The results showed that the meth-od specifically recognized CEV without cross-reaction with bovine enterovirus and bovine viral di-arrhea virus.The minimum detection limit of the method was 102.49 TCID50/mL and had good re-producibility.The prepared test strips had a shelf life of three months kept at 4 ℃.Detection of clin-ical samples using the immunochromatographic test strips showed 100%coincidence rate with RT-PCR method.In conclusion,the colloidal gold immunochromatographic test strips for detection of the emerging CEV with good specificity,sensitivity and repeatability,which provides a new techni-cal means easily used for the rapid detection/diagnosis and epidemiological investigation on CEV infection.
3.Construction of encephalitis-pathogenic NMGCF-19 E.coli strain with ompA dele-ted and its pathogenicity
Huiqi HU ; Xuyuan CUI ; Naitian YAN ; Xuebo ZHENG ; Fuhui ZHANG ; Junying HU ; Xinping WANG
Chinese Journal of Veterinary Science 2024;44(10):2123-2129
The NMGCF-19 strain is an E.coli strain isolated and identified in our laboratory from lambs manifesting severe diarrhea and meningitis.Previous analysis of the genome sequence of NMGCF-19 strain showed that the outer membrane protein A(ompA)gene was a potential viru-lent gene.In order to determine whether the ompA gene is associated with the pathogenicity of NMGCF-19 strain and the underlying mechanism,the NMGCF-19 strain with ompA knockout(NMGCF-19△ompA)was generated in this study using CRISPR/Cas9 technology and used to de-termine the role of ompA gene in mediating the encephalitis by NMGCF-19 infection and the un-derlying mechanism using the mouse model system.The results showed that the neuronal cell nec-rosis in the hippocampus in mice infected by NMGCF-19△ompA was significantly reduced and was not focal compared with that of mice infected with the wild-type NMGCF-19 strain.The number of bacteria in brain of mice infected by NMGCF-19 △ompA was significantly reduced in comparison to that of mice infected by NMGCF-19.Simultaneously,the mRNA and protein expression levels of the tight junction proteins ZO-1 and Occludin were both increased in mice infected by NMGCF-19 △ompA strain compared with the mice infected by NMGCF-19 strain.These results suggest that the ompA gene is a virulent gene and plays an important role in the invasion of the blood-brain barrier by NMGCF-19 strain in mice.

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