猪流感病毒A/Swine/Fujian/668/2001(H3N2)株感染的鸡胚尿囊液,经差速离心后,再经蔗糖密度梯度离心,提纯、纯化的猪流感病毒经NP-40处理并反复冻融,作为猪流感间接ELISA抗原,确立了间接ELISA检测方法。对29份HI试验猪流感为阴性的血清进行了检测,经统计学分析,确定间接ELISA判定标准,被检血清OD_(490nm)值≥0.20判定为阳性。该方法对猪瘟等11种猪疫病阳性血清无交叉反应,批内和批间重复试验的吸收变异系数分别在3.34%～8.12%和6.2%～9.04%之间。与HI的符合率达到92.8%,经卡方检验(P<0.01)比HI试验敏感。为猪流感抗体检测提供了快速、准确、简便的方法。
【作者单位】：中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室/兽医生物技术国家重点实验室;中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室/兽医生物技术国家重点实验室;中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室/兽医生物技术国家重点实验室;中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室/兽医生物技术国家重点实验室;中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室/兽医生物技术国家重点实验室;中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室/兽医生物技术国家重点实验室;中国农业科学院哈尔滨兽医研究所农业部动物流感重点开放实验室/兽医生物技术国家重点实验室 黑龙江 哈尔滨150001;黑龙江 哈尔滨150001;黑龙江 哈尔滨150001;黑龙江 哈尔滨150001;黑龙江 哈尔滨150001;黑龙江 哈尔滨150001;黑龙江 哈尔滨150001
【关键词】：猪流感;猪流感病毒;间接ELISA
【分类号】：S854.43
【DOI】：CNKI:SUN:ZGXQ.0.2007-04-017
【正文快照】：
　　猪流感(Swine influenza，51)即猪流行性感冒，是由正粘病毒科A型流感病毒引起的一种急性呼吸道传染病。在1918年美国首先报道发生猪流感后，1930年shope等从依阿华州分离到**株HIN一亚型猪流感病毒(swine innuenza virus，Slv)”’，后来又先后从猪体中分离到H3NZ、川NZ等各

　An indirect ELISA for detecting antibodies against swine influenza virus (SIV) was developed using antigen that was derived from allantoic fluid of chicken embryonated eggs infected with A/Swine/Fujian/668/2001 (H3N2).The antigen was purified by differential centrifugation and sucrose density gradient centrifugation,followed by NP-40 treatment and 3 freeze-thaw cycles.Samples with an optical density value≥0.2 were scored positive based on the ELISA data from twenty-nine sera that are known SIV negative in hemagglutination inhibition (HI) test.There was no cross-reactivity detected against virus strains of other 11 swine infectious diseases including classical swine fever.The variant coefficient of optical density value in a batch and be- tween batches varied from 3.34% to 8.12%,and from 6.2% to 9.04%,respectively.The coincidence rate was 92.8% between indirect ELISA and HI test.The results showed that indirect ELISA is more sensitive than HI (P<0.01) and could be used as an efficient and sensitive method to detect antibodies against SIV.