Getah virus(GETV)is an arbovirus capable of infecting humans and vertebrates such as horses and pigs via blood-sucking mosquitoes,which is extremely harmful to the livestock indus-try.Current monitoring methods are time-consuming,costly,and dependent on specialized instru-ments.These characteristics hinder rapid detection in clinical samples.Therefore,the development of a simple,rapid,specific,and sensitive method for detecting GETV antigen is crucial for the pre-vention and control of GETV.In this study,a GETV E1 monoclonal antibody strain SD17/09-E1-mAb was prepared by a prokaryotic expression system for GETV E1 protein expression,and a col-loidal gold double-antibody sandwich assay encapsulating two strains of GETV E1 monoclonal an-tibody wasestablished.The results showed that the prepared colloidal gold test strips had good sen-sitivity and did not cross-react with other common porcine virus-positive tissue samples;the test strips had a high compliance rate with the IFA assay for GETV,and could be stored at 37 ℃ for one month and at room temperature for at least three months.In this study,a colloidal gold anti-surveillance test strip for rapid detection of GETV was successfully prepared,which provides a powerful tool for GETV detection.

Getah virus(GETV)is an arbovirus capable of infecting humans and vertebrates such as horses and pigs via blood-sucking mosquitoes,which is extremely harmful to the livestock indus-try.Current monitoring methods are time-consuming,costly,and dependent on specialized instru-ments.These characteristics hinder rapid detection in clinical samples.Therefore,the development of a simple,rapid,specific,and sensitive method for detecting GETV antigen is crucial for the pre-vention and control of GETV.In this study,a GETV E1 monoclonal antibody strain SD17/09-E1-mAb was prepared by a prokaryotic expression system for GETV E1 protein expression,and a col-loidal gold double-antibody sandwich assay encapsulating two strains of GETV E1 monoclonal an-tibody wasestablished.The results showed that the prepared colloidal gold test strips had good sen-sitivity and did not cross-react with other common porcine virus-positive tissue samples;the test strips had a high compliance rate with the IFA assay for GETV,and could be stored at 37 ℃ for one month and at room temperature for at least three months.In this study,a colloidal gold anti-surveillance test strip for rapid detection of GETV was successfully prepared,which provides a powerful tool for GETV detection.

The E protein of dengue virus type Ⅱ was presented on ferritin of Helicobacter pylori to construct a novel dengue nanoparticle vaccine candidate,and the immunological indexes of the vac-cine were evaluated,aiming to provide new ideas for the development of dengue vaccine.The re-combinant plasmid of E-Ferritin was optimized and synthesized,and then transfected into HEK-293F cells.The recombinant protein was expressed,identified,purified and analyzed.Mice were im-munized with E-Ferritin nanoparticle vaccine by intramuscular injection on the hind limbs on the day 0,14 and 28.ELISA,neutralization test,flow cytometry and lymphocyte proliferation test were used to detect the levels of specific antibodies,neutralizing antibodies,CD3+,CD4+and CD8+T lymphocytes in spleen cells and the proliferation of spleen lymphocytes after specific stimulation.The target protein with a size of about 69 kDa was expressed in the cells with a single band.The purified protein concentration was 0.407 g/L,and the purity was 82.32％.The results from transmission electron microscopy showed that E-Ferritin protein could be recombined into a particle structure with a particle size of about 50 nm.The results of mouse immune experiments showed that E-Ferritin protein had good immunogenicity.The average specific antibody titer of E-ferritin protein in serum was 1∶92 160 after immunization 42 d.The main subclass of antibody was IgGl.The results of flow cytometry showed that E-Ferritin as an immunogen could induce higher levels of CD4+and CD8+T lymphocyte immune response.In lymphocyte proliferation test,the level of specific stimulation in the vaccine group was significantly higher than that in the non-specific stimulation group.In conclusion,the dengue virus envelope protein ferritin nanoparticle vaccine constructed in this study has good immunogenicity,which can provide reference for the de-velopment of new dengue vaccine candidates.

The SYBR Green Ⅰ real-time fluorescence quantitative PCR detection method was used to determine the prevalence of Japanese encephalitis virus(JEV)in mosquito vectors and cattle se-rum samples in Xinjiang.The E gene fragment of the JEV strain was amplified by PCR,cloned into a pEASY-Blunt vector,produced as a recombinant plasmid,and its sensitivity,specificity and re-producibility were verified.Between 2021 and 2022,serum samples were taken in the regions of Hami,Altay,Ili,Aksu,and Kashi in order to monitor the prevalence of JEV in livestock in Xin-jiang.The positive rate was discovered and evaluated using the established detection method.The established detection method showed a good linear relationship,and the detection interval was 4.03X102-4.03×109 copies/pL.The correlation coefficient was 0.995,the slope was-3.431,and the extreme value of the lower limit of sensitivity was 4.03 × 102 copies/pL.This method has no specific amplification for Zika virus(ZIKV)and Dengue virus(DENV).The intra group coefficient of variation of reproducibility was 0.53％-1.27％,and the inter group coefficient of variation was 0.48％-1.43％.Using this method to detect serum samples from livestock in Xinjiang from 2021 to 2022,the total positive rate was 3.28％,with positive detection rates in horses,cows,and sheep being 2.35％,6.77％,and 3.74％respectively,the virus was identified as Type Ⅰ JEV.A SYBR Green Ⅰ real-time PCR method for the detection of genotype 1 JEV was established.JEV was de-tected in the serum of horses,cattle and sheep in Xinjiang,with a total positive rate of 3.11％.

Bluetongue virus(BTV)is classified as a category Ⅱ animal epidemic disease in China,infecting multiple species and posing significant threats to the ruminant breeding industry.There are 29 serotypes of BTV,with BTV16 being one of the major serotypes currently prevalent in Chi-na.Bluetongue virus infection mainly manifests as a latent infection,making the establishment of ELISA assays crucial for epidemiological detection.In this study;the expression of the BTV16 VP2 protein was achieved using a prokaryotic expression system,and polyclonal antibodies were pre-pared using BALB/c mice.An indirect ELISA assay using VP2 protein as the encapsulated antigen was established and optimized.Clinical samples from the Guangxi Zhuang Autonomous Region were tested and analyzed for compliance with commercial kits.The results showed that the BTV16 VP2 protein was successfully expressed and purified,and the prepared polyclonal antibody exhibi-ted good immunogenicity.The ELISA assay had good specificity,with no cross-reactivity against ruminant diseases such as AKAV,FMDV and GETV.The critical values for negativity and positivity were determined to be 0.314,and the coefficients of variation(Cv)between batches and within batches were both less than 5％,indicating good reproducibility.The ELISA assay revealed a positive rate of 92.4％for 79 samples from the Guangxi Zhuang Autonomous Region,with a compliance rate of 98.7％when compared to the commercialized kit.In conclusion,this study suc-cessfully established an indirect ELISA method for BTV16,facilitating the detection of bovine clin-ical samples.

Bluetongue virus(BTV)is classified as a category Ⅱ animal epidemic disease in China,infecting multiple species and posing significant threats to the ruminant breeding industry.There are 29 serotypes of BTV,with BTV16 being one of the major serotypes currently prevalent in Chi-na.Bluetongue virus infection mainly manifests as a latent infection,making the establishment of ELISA assays crucial for epidemiological detection.In this study;the expression of the BTV16 VP2 protein was achieved using a prokaryotic expression system,and polyclonal antibodies were pre-pared using BALB/c mice.An indirect ELISA assay using VP2 protein as the encapsulated antigen was established and optimized.Clinical samples from the Guangxi Zhuang Autonomous Region were tested and analyzed for compliance with commercial kits.The results showed that the BTV16 VP2 protein was successfully expressed and purified,and the prepared polyclonal antibody exhibi-ted good immunogenicity.The ELISA assay had good specificity,with no cross-reactivity against ruminant diseases such as AKAV,FMDV and GETV.The critical values for negativity and positivity were determined to be 0.314,and the coefficients of variation(Cv)between batches and within batches were both less than 5％,indicating good reproducibility.The ELISA assay revealed a positive rate of 92.4％for 79 samples from the Guangxi Zhuang Autonomous Region,with a compliance rate of 98.7％when compared to the commercialized kit.In conclusion,this study suc-cessfully established an indirect ELISA method for BTV16,facilitating the detection of bovine clin-ical samples.