中国动物检疫
中國動物檢疫
중국동물검역
CHINA ANMAL QUARANTINE
2012年
4期
37-40
,共4页
董志珍%肖妍%赵祥平%栾慎顺%张霞
董誌珍%肖妍%趙祥平%欒慎順%張霞
동지진%초연%조상평%란신순%장하
单克隆抗体%McAb-ELISA%非洲猪瘟%实验诊断
單剋隆抗體%McAb-ELISA%非洲豬瘟%實驗診斷
단극륭항체%McAb-ELISA%비주저온%실험진단
monoclonal antibody%McAb-ELISA%African swine fever: laboratory diagnosis
用基因重组技术制备的非洲猪瘟蛋白P54免疫BALB/C小鼠,将免疫鼠的脾细胞与SP2/0骨髓瘤细胞融合,采用有限稀释法进行克隆,获得能稳定分泌抗ASFV单克隆抗体的杂交瘤细胞株。使用该单克隆抗体,建立了检测血清中非洲猪瘟抗体的竞争法ELISA。实验结果表明:ELISA竞争法特异性高,无交叉反应,灵敏度高于间接免疫荧光法,可用于猪血清的非洲猪瘟抗体检测。该法的建立对非洲猪瘟实验诊断的标准以及流行病学调查具有重要的现实意义。
用基因重組技術製備的非洲豬瘟蛋白P54免疫BALB/C小鼠,將免疫鼠的脾細胞與SP2/0骨髓瘤細胞融閤,採用有限稀釋法進行剋隆,穫得能穩定分泌抗ASFV單剋隆抗體的雜交瘤細胞株。使用該單剋隆抗體,建立瞭檢測血清中非洲豬瘟抗體的競爭法ELISA。實驗結果錶明:ELISA競爭法特異性高,無交扠反應,靈敏度高于間接免疫熒光法,可用于豬血清的非洲豬瘟抗體檢測。該法的建立對非洲豬瘟實驗診斷的標準以及流行病學調查具有重要的現實意義。
용기인중조기술제비적비주저온단백P54면역BALB/C소서,장면역서적비세포여SP2/0골수류세포융합,채용유한희석법진행극륭,획득능은정분비항ASFV단극륭항체적잡교류세포주。사용해단극륭항체,건립료검측혈청중비주저온항체적경쟁법ELISA。실험결과표명:ELISA경쟁법특이성고,무교차반응,령민도고우간접면역형광법,가용우저혈청적비주저온항체검측。해법적건립대비주저온실험진단적표준이급류행병학조사구유중요적현실의의。
The P54 protein of African swine fever virus(ASFV) was prepared with gene recombination technique and used to immune Balb/C mice, the spleen cells of immunized mice was fused with SP2/0, cloned by limiting dilution thus obtaining hybridoma cell lines secreting anti-ASFV antibody stably. An ELISA for the detection of ASFV was established using the monoclonal antibody. The results showed that the ELISA kit could be used for the detection of African swine fever antibody with high specificity and no cross-reaction. The development of the ELISA method had important practical significance for the laboratory diagnosis and epidemiological investigation of ASFV.