牛传染性鼻气管炎病毒抗体胶体金检测试纸条的制备
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摘要
采用SF 9昆虫细胞-杆状病毒系统重组表达牛传染性鼻气管炎病毒gD蛋白抗原,经纯化鉴定后将gD蛋白用胶体金标记作为示踪抗原,未标记的gD抗原作为捕获抗原,并以羊抗牛IgG抗体作为质控抗体,建立了牛传染性鼻气管炎病毒双抗原夹心法胶体金检测试纸条。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、牛病毒性腹泻病毒等阳性血清无交叉反应。使用建立的胶体金试纸条和IDEXX牛传染性鼻气管炎病毒抗体ELISA检测试剂盒同时检测112份牛血清,阳性符合率为93.5%,阴性符合率为96.0%,总符合率为94.6%。说明该试纸条可以应用于临床牛传染性鼻气管炎病毒抗体的诊断和检测。
gD protein antigen of infectious bovine rhinotracheitis virus was prepared by constructing the recombinant expression with sf9 insect cell-baculovirus system, and the purified gD protien was labeled as a tracer by colloidal gold. The colloidal gold test strip of detecting the antibody of infectious bovine rhinotracheitis virus was established with gD 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 high sensitivity, strong specificity, and no cross-reactivity towards bovine foot-and-mouth disease virus and bovine viral diarrhea virus. 112 bovine serum samples were detected by the strip and ELISA(IDEXX-IBRV Antibody Detection Kit) was applied as the comparative method, the coincidence rate of positive results was 93.6%, the coincidence rate of negative results was 96.0%, and the total coincidence rate was 94.6%. This strip could be used for the diagnosis and antibody detection of infectious bovine rhinotracheitis virus.
gD protein antigen of infectious bovine rhinotracheitis virus was prepared by constructing the recombinant expression with sf9 insect cell-baculovirus system, and the purified gD protien was labeled as a tracer by colloidal gold. The colloidal gold test strip of detecting the antibody of infectious bovine rhinotracheitis virus was established with gD 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 high sensitivity, strong specificity, and no cross-reactivity towards bovine foot-and-mouth disease virus and bovine viral diarrhea virus. 112 bovine serum samples were detected by the strip and ELISA(IDEXX-IBRV Antibody Detection Kit) was applied as the comparative method, the coincidence rate of positive results was 93.6%, the coincidence rate of negative results was 96.0%, and the total coincidence rate was 94.6%. This strip could be used for the diagnosis and antibody detection of infectious bovine rhinotracheitis virus.
引文
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[11] 周泰冲,叶章明,黎少权,等.从新西兰进口奶牛中分离传染性牛鼻道气管炎病毒[J].兽医科技杂志,1981(1):8-11.
[12] 费恩阁,李德昌,丁壮.动物疫病学[M].北京:中国农业出版社,2004.
[13] 丁国伟,李琛,李玉安,等.牛传染性鼻气管炎病毒gD蛋白的原核表达及其免疫原性[J].湖南农业大学学报(自然科学版),2018,44(1):8-11.
[2] 李树博.牛传染性鼻气管炎病毒的研究进展[J].畜牧兽医科技信息,2015(7):73-73.
[3] 季新成.牛传染性鼻气管炎病毒和牛病毒性腹泻病毒分子生物学检测技术研究[D].杨凌:西北农林科技大学,2010.
[4] 牛国辉,季新成,段晓东,等.牛病毒性腹泻病毒的分离与鉴定[J].新疆农业科学,2010,47(5):986-991.
[5] 徐晓琴,冷雪,李真光,等.牛传染性鼻气管炎病毒IBRV-LN01/08株的分离鉴定[J].中国兽医科学,2010,40(4):352-356.
[6] 朱远茂,王海燕,薛飞,等.牛传染性鼻气管炎间接ELISA诊断方法的建立[J].中国兽医科学,2005,35(12):959-963.
[7] 陈婷.牛传染性鼻气管炎病毒gD基因原核表达及间接ELISA检测方法的建立[D].杨凌:西北农林科技大学,2012.
[8] 徐淑菲,孔繁德,周斌华.牛传染性鼻气管炎PCR检测方法的建立[J].中国动物检疫,2006,23(11):20-23.
[9] 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.
[10] Shuaib M,Green C,Rashid M,et al.Herd risk factors associated with sero-prevalence of Maedi-Visna in the Manitoba sheep population[J].Can Vet J,2010,51(4):385-390.
[11] 周泰冲,叶章明,黎少权,等.从新西兰进口奶牛中分离传染性牛鼻道气管炎病毒[J].兽医科技杂志,1981(1):8-11.
[12] 费恩阁,李德昌,丁壮.动物疫病学[M].北京:中国农业出版社,2004.
[13] 丁国伟,李琛,李玉安,等.牛传染性鼻气管炎病毒gD蛋白的原核表达及其免疫原性[J].湖南农业大学学报(自然科学版),2018,44(1):8-11.