摘要
为快速、便捷地检测A型口蹄疫病毒(FMDV)抗体,将胶体金颗粒与口蹄疫A型疫苗株146S病毒粒子(FMDV-146S)结合,作为金标抗原(g FMDV-146S),与其抗体(FMDV-146S-Ab)分别作为检测带(T带)与质控带(C带)组装试纸条,通过检测已知各型FMDV阴性、阳性血清,水疱性口炎(VS)、猪水泡病(SVD)和羊传染性脓疱(CE)的阳性血清,验证其敏感性、特异性与稳定性;同时平行用液相阻断ELISA(LPB-ELISA)方法进行对比检验。结果显示,该试纸条检测抗体效价最低达1∶128;SVD、VS、CE阳性血清及FMDV阴性血清的检测结果均为阴性,A型FMDV阳性血清100%显阳性,O型和Asia 1型的FMDV阳性血清仅出现10%的阳性率;该试纸条于4℃保存10个月,不同批次间、同一批次内的重复性检测结果差异不显著(P>0.01);与LPB-ELISA检测结果的相关性良好,以此建立该试纸条的半定量比色卡。本试验制备的A型口蹄疫抗体金标检测试纸条具有较好的灵敏度、特异性、稳定性,能够准确评价A型口蹄疫疫苗的免疫效果。
For rapid detecting antibodies against serotype A,foot-and-mouth disease virus（FMDV）,a gold-labeled immunochromatographic test strip（shortened as gold test strip） was developed in present study,with purified 146 S particles of FMDV serotype A（FMDV-146S） coupled with colloidal gold（g FMDV-146S） as test line（T）and the antibodies against FMDV-146S（FMDV-146S-Ab） as control line（C）.The sensitivity,specificity and stability of this gold test strip were assessed by testing positive and negative FMDV sera and positive sera against VS,SVD and CE.Parallel tests of the positive FMDV sera by both gold test strip and LPB-ELISA were performed.The present prepared gold test strip showed its sensitivity as 1 ∶ 128.Of specificity,no cross positive reactions were found with VSV,SVDV,CEV positive and FMDV negative sera.All FMDV positive sera showed 100% positivity.However 10% of cross-positive were observed between FMDV serotype Asia I and serotype O.Of stability,the gold test strip remained effective after ten-month storation at 4 ℃.Three batches were detected in triplicate,with no significant difference（P〉0.01）.The gold test strip and LPB-ELISA shared a high coincidence rate,so a color card for semi-quantization of antibodies against FMDV was established.The present gold test strip demonstrated high sensitivity,specificity and stability and its reliability for evaluating the efficacy of vaccination against FMDV serotype A.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2016年第9期1123-1127,共5页
Chinese Veterinary Science
基金
国家自然科学基金项目(31260616)
甘肃省生物技术研究与应用项目(GNSW-2014-8)
关键词
口蹄疫病毒
胶体金
免疫层析试纸
foot-and-mouth disease virus（FMDV）
colloidal gold
immunochromatographic test strip
