应用单克隆抗体建立夹心ELISA检测EDS—76病毒

Establishment of a Sandwich Using Monoconal Antibody ELISA Test for the Detection of EDS- 76 Virus

利用单克隆抗体建立夹心ＥＬＩＳＡ方法，对人工感染鸡带毒和排毒检测，表明第３天血、脾、输卵管带毒，第８天时各脏器几乎都带毒，第１４天仅输卵管、肾、肝检出病毒。第６天时鸡蛋带毒，泄殖腔排毒；直至１４天时仍带毒和排毒。夹心ＥＬＩＳＡ与ＨＡ、鸭胚接种相比较，在３６份样品中，ＨＡ检出１９份。ＥＬＩＳＡ检出２４份，鸭胚接种检出１２份。病毒在鸭胚和细胞上复制动态表明，２４小时后血凝试验和夹心ＥＬＩＳＡ都能检出病毒，随着时间延长，ＨＡ价和ＥＬＩＳＡ价（ＰＮ）继续上升，８４小时至９６小时到最高峰。在２４小时时，无血凝性，但ＥＬＩＳＡ检测为阳性，因此夹心ＥＬＩＳＡ可用于病毒检测。单抗夹心ＥＬＩＳＡ的建立为生产上提供了一种更简单、快速、灵敏、应用广泛的病毒检测方法。

EDS-76 viruses carried and discharged in experimental infected chickenswere detected by a monoclonal antibody- based Sandwich Enzyme- Linked ImmunosoubentAssay(S-ELISA).The results indicated that there were viruses in the blood,spleen andoviduct on day 3 postinfectin(PI),and viruses observed in all viscera on day 8 PI,but virus-es were detected only in the oviduct,kidney and liver on day 14PI.Beginning on day 6 PI,viruses in eggs were detected and viruses in chickens were discharged through the cloaca,however,viruses in eggs could be detected and viruses in chickens could be discharged untilthe end of the study,dav 14 PI. When S-ELISA was compared with,HA test and duck em-bryo- inoculation assay,there were 19 positive samples of 36 samples by the HA test,24 pos-itive samples by S-ELISA and 12 positive samples by duck embryo-inoculation assay. Repro-ductive development of EDS-76 viruses grown in duck embryo and duck embryo fibroblastcells(DEFC)suggested that EDS-76 viruses could be detected by both the HA test and S-ELISA after 24 hours PI.The tres of the HA test and the P/N values of S-ELISA contin-ued to increase as the time of infection was extended,and reached a maximum at 84 to 96hours PI.But there weren′t viruses detected bv the HA test in dukc embryo and DEFCs at24 bours PI,however viruses could be detected bv S-ELISA,Therefore,S-ELISA could beused to detect EDS-76 virus. Establishment of a monoclonal antibody-based S-ELISA pro-vided a simple,fast,sensitive and universally used method for the diagnosis of EDS-76