牛病毒性腹泻病毒抗体胶体金检测试纸条的制备
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摘要
采用sf9昆虫细胞-杆状病毒系统重组表达牛病毒性腹泻病毒P80蛋白抗原,经纯化鉴定后将P80蛋白用胶体金标记作为示踪抗原,未标记的P80抗原作为捕获抗原,并以羊抗牛IgG抗体作为质控抗体,建立了牛病毒性腹泻病毒抗体快速诊断试纸条。试验结果表明,该试纸条检测灵敏度高,特异性良好,与牛口蹄疫病毒、牛传染性鼻气管炎病毒无交叉反应。使用建立的胶体金试纸条和IDEXX牛病毒性腹泻病毒ELISA抗体检测试剂盒同时检测80头份牛血清,阳性结果符合率为86.7%,阴性结果符合率为100%,总符合率为95%。该试纸条可用于临床牛病毒性腹泻病毒抗体的诊断和检测。
P80 protiein antigen of bovine viral diarrhea virus was prepared by constructing the recombinant expression with sf9 insect cell-baculovirus system, and the purified P80 antigen was labeled as a tracer by colloidal gold. The colloidal gold test strip of detecting the antibody of bovine viral diarrhea virus was established with P80 protein as a captured antigen, and with the sheep-anti-bovine antibody as a quality control antibody. The results showed that this strip was with highly sensitive detection, strong specificity, and no cross-reactivity with bovine foot-and-mouth disease virus and infectious bovine rhinotracheitis virus. 80 bovine serum samples were detected by the strip and ELISA(IDEXX-BVDV P80 Antibody Detection Kit) wass applied as the comparative method, the coincidence rate of positive results was 86.7%. The coincidence rate of negative results was 100%, the total coincidence rate was 95%. This strip could be used for the diagnosis and antibody detection of bovine viral diarrhea virus.
P80 protiein antigen of bovine viral diarrhea virus was prepared by constructing the recombinant expression with sf9 insect cell-baculovirus system, and the purified P80 antigen was labeled as a tracer by colloidal gold. The colloidal gold test strip of detecting the antibody of bovine viral diarrhea virus was established with P80 protein as a captured antigen, and with the sheep-anti-bovine antibody as a quality control antibody. The results showed that this strip was with highly sensitive detection, strong specificity, and no cross-reactivity with bovine foot-and-mouth disease virus and infectious bovine rhinotracheitis virus. 80 bovine serum samples were detected by the strip and ELISA(IDEXX-BVDV P80 Antibody Detection Kit) wass applied as the comparative method, the coincidence rate of positive results was 86.7%. The coincidence rate of negative results was 100%, the total coincidence rate was 95%. This strip could be used for the diagnosis and antibody detection of bovine viral diarrhea virus.
引文
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[3] 童钦,霍志云,胡桂学,等.牛病毒性腹泻病毒检测方法的研究进展[J].中国农学通报,2012,28(14):79- 83.
[4] 寇改霞,陈茂盛,吕康年.奶牛病毒性腹泻-黏膜病的病原分离及荧光抗体诊断[J].中国兽医科技,1992,22(9)∶22- 23.
[5] 黄骏明,郭连喜,王燕昌,等.牛病毒性腹泻诊断方法的研究Ⅰ.应用直接免疫荧光抗体技术检查牛鼻上皮细胞中的牛病毒性腹泻病毒[J].家畜传染病,1986,6(3):28- 31.
[6] 王新平,程焰.双抗体夹心阻断ELISA检测血清中牛病毒性腹泻-粘膜病病毒抗体的研究[J].中国兽医学报,1993,13(4):334- 338.
[7] 范学政,宁宜宝,王琴,等.用RT-PCR方法检测猪瘟细胞苗中污染牛病毒性腹泻病毒[J].中国兽医杂志,2010,46(1):8- 10.