中国兽医科学2025，Vol.55Issue(5)：625-631,7.DOI:10.16656/j.issn.1673-4696.2025.0083
A型塞内卡病毒的分离鉴定及3A蛋白的多克隆抗体制备
Isolation of senecavirus A and preparation of anti-3A protein polyclonal antibody
摘要
Abstract
Senecavirus A(SVA),as a foreign pathogen that can cause vesicular symptoms in pigs,is difficult to distinguish from clinical symptoms caused by foot-and-mouth disease virus.In this study,we successfully isolated one SVA isolate and named it HN19.By whole genome sequencing and genetic evolution analysis,it was found that the isolate displayed the highest similarity with the CH-HN-2017 strain isolated from China.The HN2019 strain can effectively replicate and proliferate in porcine kidney passage cells(PK-15)and hamster kidney cells(BHK-21),and can produce cytopathic effect after virus infection.The viral growth characteristics were analyzed by the experiments such as one-step virus growth curve,indirect immunofluorescence,and Western-blot.Meanwhile,a prokaryotic expression vector expressing the nonstructural protein 3A was constructed using the pCold-TF plasmid and transformed into an Escheri-chia coli system.It was found that the recombinant protein could be expressed soluble in the supernatant.The purified recombinant protein was immunized four times in BALB/c mice to successfully prepare polyclonal antibodies against 3A protein.The polyclonal antibody titer was 1∶128 000.The antibody can specifically recognize SVA-infected cells to produce specific fluorescence,demonstrating its good reactivity and specificity.This study provides an effective tool for further studying the biological function of 3A protein and the pathogenic mechanism of SVA.关键词
A型塞内卡病毒/遗传进化分析/3A蛋白/多克隆抗体Key words
senecavirus A/genetic evolution analysis/3A protein/polyclonal antibody分类
农业科技引用本文复制引用
韩松,童光志,李丽薇,段宏勇,杨楠,王树茂,高飞,赵款,周艳君,童武,姜一峰..A型塞内卡病毒的分离鉴定及3A蛋白的多克隆抗体制备[J].中国兽医科学,2025,55(5):625-631,7.基金项目
上海市科技兴农项目(2022-02-08-00-12-F01162) （2022-02-08-00-12-F01162）
国家重点研发计划项目(2022YFD1800305) （2022YFD1800305）
中国农业科学院科技创新工程项目(CAAS-CSLPDCP-202301) （CAAS-CSLPDCP-202301）
