作 者：
杨振兴;朱建波;肖雷;高林;苗海生;何于雯;谢佳芮;杨恒;李华春
单 位：
云南省畜牧兽医科学院云南省热带亚热带动物病毒病重点实验室
摘 要：
为建立一种简便、快速的中山病血清抗体监测方法,本研究采用纯化的中山病病毒(CHUV)包被ELISA反应板,兔抗CHUV多克隆抗体作为竞争抗体,建立了检测CHUV抗体的竞争ELISA(C-ELISA)方法。结果显示:抗原最佳包被浓度为4.12μg/m L,竞争抗体最佳使用浓度为1∶12 000倍。特异性试验显示该C-ELISA方法仅能检测CHUV抗体,其它相关病毒阳性血清检测结果均为阴性。对已知阳性样品的检测结果表明,本研究建立的C-ELISA方法敏感性高于血清中和试验和琼脂扩散试验。分别用C-ELISA、RT-PCR和病毒分离试验对监控动物每周采集的血清和抗凝血样品进行检测,ELISA结果与其它两种方法检测结果对应一致。本研究建立的C-ELISA为CHUV抗体的检测提供了一种快速、方便、可靠的方法。
译 名：
Establishment of a competitive ELISA for detecting antibodies against Chuzan disease virus
作 者：
YANG Zhen-xing;ZHU Jian-bo;XIAO Lei;GAO Lin;MIAO Hai-sheng;HE Yu-wen;XIE Jia-rui;YANG Heng;LI Hua-chun;Yunnan Veterinary and Animal Science Institute;
关键词：
Chuzan disease virus;;multi-clone antibody;;competitive ELISA
摘 要：
In order to establish a convenient and rapid method for monitoring serum antibody against Chuzan disease virus(CHUV), a competitive ELISA(C-ELISA) was established using purified viral protein as coating antigen and rabbit polyclonal antibody as competitive antibody. The results showed that the optimal concentration of antigen was 4.12μg/m L and the optimal concentration of antibody was 1 ∶12,000. Specific tests showed that this method specifically detect the antibodies against CHUV.The detection results of known positive samples showed that the sensitivity of C-ELISA established in this study was higher than that of the serum-neutralization test and agar gel immunodiffusion test. The clinical samples collected from monitoring animals were detected by the C-ELISA, RT-PCR and virus isolation, respectively. The results of the C-ELISA were consistent with those of the other two methods. The C-ELISA established in this study provides a rapid, convenient and sensitive method for detecting the antibodies against CHUV.
