“全胚双氟碳法”制备禽脑脊髓炎琼扩抗原的研究

用禽脑脊髓炎病毒（AEV）Van Rockel毒株接种6日龄SPF鸡胚，感染11 d后，分别收集鸡胚、绒毛尿囊膜及尿囊液、鸡胚剔除羽毛、眼球、爪和喙后与绒毛尿囊膜一起，制成匀浆，反复冻融3次，通过2次氟碳（三氯三氟乙烷）从全胚匀浆中萃取制备AE琼扩抗原（AGP－Ag)用进口标准AE AGP试剂进行标化，结果发现：制备的AD AGP－Ag与标准阳性血清之间的清晰、致密的沉淀线，并与标准AE AGP－Ag的沉淀线完全吻合，表明两者的沉淀反应一致，从而了AE AGP－Ag制备新方法－“全胚双氟碳法”其制备AE AGP－Ag有如下特点：①产量高，平均每3个SPF鸡可获得1mL AE AGP－Ag; ②特异，与类症阳性血清无交叉反应；③敏感，AGP试验中毋需重复加样，普通琼脂即可出线；④稳定性好，保存期长；⑤与标准AE AGP－Ag阳性检出符合率均达100％。本研究表明用“全胚双氟碳法”制备的AEAGP－Ag达到了进口标准，AEAGP－Ag质量，可用于SPF鸡AE抗体的监测，且生产成本低，也可用于商品鸡AEV感染的检测。

Preparation of theAgar-Gel-Precipitating Antigen for Avian Encephalomyelitis by Double FluorcarbanExtractions from Whole Embryo

Six-day-old SPF chicken embryos were inoculated with Avian Encephalomylitis Virus (AEV) Van Rockel,and the whole embryos (without feathers,eyes,claws and beaks) were collected and homogenized at 11 days after inoculation.The homogenate was frozenthawed for 3 times,and extracted twice with Fluorocarban.The extracted sample was ultracentrifugated at 7×104　g.min-1 for 3 h to obtain the concentrated AE AGP-Ag .The Ag was evaluated with the standard AE AGP-reagents.The results showed that there was clear precipitation line between the Ag and the standard positive serum,and the line matched with the standard Ag's line exactly,which suggested that the two kinds of Ag reacted in the same manner.The Ag output is higher and the Ag is specific and sensitive.　1 mL Ag could be producted with 3 SPF chicken embryos. The Ab detection rate of the Ag was identified with that of the standard Ag in monitoring SPF chickens and immunized or infected chickens.This revealed that the prepared Ag can be used to monitor AE-Ab of SPF chickens,immunized breeder flocks and infected commercial chickens.