摘要
从英国中心兽医实验室（ＣＶＬ）引进禽流感病毒１４种血凝素亚型（Ｈ１～Ｈ１４）的标准毒株，经９～１１日龄的ＳＰＦ鸡胚增殖、传代后收取感染鸡胚的羊水及尿囊液，经１０００ｘｇ离心１５分钟后，收取上清作为免疫原。取８只１２周龄ＳＰＦ鸡分成Ａ、Ｂ两组，进行人工感染鸡的抗体消长规律试验。选用Ｈ３毒株制备的免疫原，按１ｍｌ／只的剂量翅静脉接种Ａ、Ｂ两组的８只ＳＰＦ鸡，１周后Ｂ组的４只鸡复免一次，分别用ＨＩ和ＡＧＰ试验测定２种方法免疫鸡的抗体水平．监测结果表明：经１次免疫的鸡（Ａ组）于免疫后１５天ＨＩ抗体升至最高，以后抗体逐渐下降，且产生的抗体个体差异不大，ＡＧＰ试验结果只有于免疫后２０～５５天内全部为阴性；经重复免疫的鸡（Ｂ组），于初免后１５～４５天内ＨＩ抗体都维持较高水平，且产生的抗体个体差异不大，ＡＧＰ试验结果在初免疫后１５７５天内全部为阳性。据此，我们根据禽流感病毒株毒力的强弱（参照其ＥＩＤ５０，用纯化后的鸡胚尿囊液制备的免疫原，按０５～２５ｍｌ／只的剂量分别免疫１２周龄ＳＰＦ鸡，１周后加强免疫一次于二免后１０～１５天放血，制备出禽流感病毒１４个血凝素亚型的分型血清，并有霍乱滤液（ＲＤＥ）法对血清进行了处理。
In this study,2 different vaccination procedures for producing antibody to hemagglutinin of avian influenza virus(AIV) were evaluated,then hemagglutininsubtyping sera were prepared in chickens.Firstly,specific pathogen free(SPF) chickens were infected with AIV A/Duck/Ukraine/1/63(113) strain through 2 different procedures (single vaccination and twice vaccination),and the hemagglutinationinhibition (HI) antibody levels were investigated.The results showed that the twice vaccination procedure indure induced hiigher titer of HI antibody which lasted much longer as well.After that,SPF chickens were inoculated with different AIVs through the twice vaccination procedure.Hemagglutininsubtyping sera specific to H1H14 subtypes were prepared from the birds,and then processed with receptor destroying enzyme to remove nonspecific inhibition factors in the sera.
关键词
禽流感病毒
血凝素
分型血清
抗体
Avian influenza virus Hemagglutinin Subtyping serum
