Establishment and preliminary application of prokaryotic expression of BTV16 re-combinant VP2 protein,preparation of polyclonal antibody and indirect ELISA detection method
10.16303/j.cnki.1005-4545.2024.12.07
- VernacularTitle:BTV16重组VP2蛋白原核表达、多克隆抗体制备及间接ELISA检测方法的建立与初步应用
- Author:
Mingzhu ZHANG
1
;
Peng WANG
;
Jiaxin TIAN
;
Shigang CHEN
;
Junduo BAO
;
Xiangshu QIU
;
Huijun LU
;
Chang LI
Author Information
1. 吉林农业大学动物医学院,吉林长春 130118;中国农业科学院长春兽医研究所,吉林长春 130122
- Publication Type:Journal Article
- Keywords:
prokaryotic expression;
polyclonal antibody;
indirect ELISA;
BTV
- From:
Chinese Journal of Veterinary Science
2024;44(12):2549-2555
- CountryChina
- Language:Chinese
-
Abstract:
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.
