摘要
以犬冠状病毒(CCV)TN449株为对象,对CCV核衣壳蛋白(N蛋白)编码基因进行克隆和原核表达,通过使用纯化的重组N蛋白,建立了CCV抗体间接ELISA检测方法。结果显示,重组N蛋白具有良好的CCV抗原反应性;经优化ELISA反应条件,确定重组N蛋白最佳包被浓度为1.0μg/m L、待检血清的最佳浓度为1∶100、阴阳性判定临界值为0.418;通过对比检测犬瘟热等多种犬病阳性血清,未发生交叉反应,证实该间接ELISA方法特异性好;用建立的间接ELISA方法对本室保存的50份CCV阳性血清和阴性血清进行复检,证实其符合率为100%。
The gene of nucleocapsid protein（N protein）from CCV strain TN449 was cloned and expressed in Escherichia coli.Using the purified N protein,an indirect ELISA method for CCV detection was established.The results showed that the purified re-combinant N protein had antigen reactivity of CCV.The reaction condition was optimized,including 1.0 μg/m L coating antigen withpurified reconbinant N protein,1∶100 dilution of testing serum,cut-off value of 0.418（OD450nm）. The specific tests showed thatthere was no cross-reaction to the anti-sera against other canine diseases such as canine distemper.The 50 samples of CCV-posi-tive and negative sera were used to test the stability and reliability of the indirect ELISA,and the coincidence rate of positve andnegative results were 100%.
出处
《中国兽医杂志》
CAS
北大核心
2015年第8期86-88,92,共4页
Chinese Journal of Veterinary Medicine
基金
上海市科委科研项目(11140901100)
