摘要
以偶联多肽为抗原建立检测O型猪口蹄疫病毒抗体的ELISA方法。通过FMDV多肽序列的筛选、化学合成及反应原性鉴定,将多肽与非蛋白结构多聚体载体进行偶联。利用偶联多肽作为抗原,通过方阵滴定确定偶联多肽、血清及HRP的最佳工作浓度,确定方法的阴阳性临界值,然后进行特异性试验、敏感度测定及临床猪血清样本的免疫效果评估等。结果表明,偶联多肽具有良好的反应原性,最佳包被浓度为1.0μg/mL,血清最佳稀释度为1:32,HRP最佳稀释度为1:8 000,方法特异性好,敏感度高,操作方便省时,能够对FMDV的自然感染进行检测,也可对FMDV免疫后的猪血清抗体水平进行评估。
An ELISA using conjugated peptides as antigen was established to detect swine antibodies of Foot-and-Mouth virus Type O. In order to conjugate the peptides to a non-protein carrier, the FMDV peptide sequences selected by bioinforulatics, chemical synthesis and immunity identification were done. Using the conjugated peptides as antigen for the indirect ELISA, the optimal concentration of conjugated peptides, the sera as well as the HRP were detected by phalanx titrations. The critical value of negative and positive controls, the specificity and sensitivity of this method were confirmed. The immune efficacies of clinical samples of swine sera were assessed. The results showed that the conjugated pcptides as the antigen had effective immunity and the optimal coating concentration was 1.0 μg/mL, the optimal dilutions of the positive control serum and the HRP were 1：32 and 1：8 000, respectively. The method has high specificity and sensitivity. It is easy to operate and saves time. This method could be used not only to detect sera of FMDV infected pigs, but also to assess the antibody levels of animals immunized by FMDV.
出处
《广东农业科学》
CAS
CSCD
北大核心
2014年第4期136-139,155,共5页
Guangdong Agricultural Sciences
基金
广东省兽用基因工程疫苗和诊断试剂研发产学研示范基地(2011B090500021)
广东省科技型中小企业创新专项资金项目(2012CY005)
