扬州大学学报（农业与生命科学版）2023，Vol.44Issue(6)：77-83,7.DOI:10.16872/j.cnki.1671-4652.2023.06.010
非洲猪瘟病毒p30二聚体鉴定及胶体金免疫层析方法建立
Identification of African swine fever virus p30 dimer and development of a colloidal gold immunochromatographic method for detecting it
摘要
Abstract
To establish a simple preparation and sensitive method for the rapid detection of African swine fever virus(ASFV)antigen,the recombinant protein p30 with His tag and Flag tag was co-expressed in 293A cells using the expres-sion vector pIRES.After lysing the cells,p30 was found to be a polymer by immunoprecipitation.The relative molecular mass of purified p30 was further determined by mass spectrometry,and the results confirmed that ASFV p30 could form dimers spontaneously.With that,a colloidal gold immunochromatographic method was established for the detection of p30 dimers by a single monoclonal antibody 6H9A10 against p30,and the ASFV p30 recombinant antigen at the concen-tration of 2.1 ng·mL-1 could be detected instead of the viral antigens,such as antigens of PRV,CSFV,PRRSV,PCV-2,PEDV,PPV and SV A.100 clinical samples including 20 ASFV negative sera,20 ASFV negative tissues,10 AS-FV negative plasma,21 ASFV positive sera,21 ASFV positive tissues,and 8 ASFV positive plasma were tested by the immunochromatographic method,the results showed that the 48 negative samples and 49 positive samples detected were consistent with known samples.This study indicates that the established colloidal gold immuno-chromatographic method for the detection of p30 by single monoclonal antibody sandwich has good specificity and sensitivity,and is expected to be used for the rapid clinical detection of ASFV antigen.关键词
非洲猪瘟病毒/p30/二聚体/胶体金免疫层析方法Key words
African swine fever virus/p30/dimer/colloidal gold immunochromatographic method分类
农业科技引用本文复制引用
张胜楠,周雅维,张子羿,单玉平,夏晓莉,张鑫宇..非洲猪瘟病毒p30二聚体鉴定及胶体金免疫层析方法建立[J].扬州大学学报（农业与生命科学版）,2023,44(6):77-83,7.基金项目
国家"十四五"重点研发项目(2021YFD1801202) （2021YFD1801202）
上海市"科技创新行动计划"农业科技领域项目(20392002700) （20392002700）
连云港市第六期"521高层次人才培养工程"科研项目(LYG06521202204) （LYG06521202204）
