检测猪流行性腹泻病毒S1蛋白抗体的间接ELISA方法

发布时间：2018-03-07 11:46

本文选题：猪流行性腹泻病毒　切入点：重组PEDV　出处：《畜牧兽医学报》2017年06期 　论文类型：期刊论文

【摘要】：为了建立检测猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)抗体的间接ELISA方法,初步分析PED灭活疫苗免疫母猪所产仔猪的血清母源特异抗体动态,笔者应用重组PEDV部分纤突(spike,S)蛋白作为检测抗原,建立了基于PEDV S1蛋白的间接ELISA抗体检测方法,并用该方法检测了产前4周用PED灭活疫苗免疫母猪的初乳、分娩当天血清及其仔猪1、7、14、21、28和35日龄(d)时血清中的PEDV抗体。结果显示:建立的间接ELISA方法能够特异地检测PEDV抗体,与猪瘟病毒、猪圆环病毒2型、猪繁殖与呼吸综合征病毒、伪狂犬病病毒、猪传染性胃肠炎病毒及猪轮状病毒的抗血清无交叉反应;敏感性高于免疫过氧化物酶单层试验(immunoperoxidase monolayer assay,IPMA),与IPMA检测结果的符合率为97.14%;批内和批间重复性试验的变异系数小于10%。免疫母猪初乳、分娩当天血清中的PEDV抗体阳性率为100%,初乳比血清中的平均抗体水平低;1d仔猪血清PEDV抗体阳性率为50%,平均抗体水平低,与初乳中的抗体水平接近,7d以后所有仔猪血清全部转为PEDV抗体阴性。建立了检测PEDV抗体的间接ELISA方法;产前用PEDV灭活疫苗给怀孕母猪免疫1次时,其所产仔猪血清中的母源特异抗体阳性率和抗体水平低,且维持时间短暂。
[Abstract]:In order to establish an indirect ELISA method for detection of porcine epidemic diarrhea virus (PEVV) antibody in porcine epidemic diarrhea virus, the maternal specific antibody dynamics of piglets immunized with PED inactivated vaccine were preliminarily analyzed. The recombinant PEDV partial spike-Sv protein was used as the detection antigen. The indirect ELISA antibody detection method based on PEDV S1 protein was established, and the colostrum of sows immunized with PED inactivated vaccine at 4 weeks before birth was detected by this method. The results showed that the indirect ELISA assay could specifically detect PEDV antibody against swine fever virus, porcine circovirus type 2, porcine reproductive and respiratory syndrome virus. The antiserum of pseudorabies virus, porcine infectious gastroenteritis virus and porcine rotavirus had no cross reaction. The sensitivity was higher than that of immunoperoxidase monolayer assayma, and the coincidence rate with the results of IPMA detection was 97.140.The coefficient of variation of repeated tests was less than 10.The immune sow colostrum was immunized. The positive rate of PEDV antibody in serum was 100 on the day of delivery, and the positive rate of serum PEDV antibody in colostrum was lower than the average antibody level in serum. The positive rate of PEDV antibody in serum of piglets on day 1 was 50 and the average antibody level was lower. The antibody level in colostrum was close to that in colostrum and all piglets turned to PEDV antibody negative after 7 days. An indirect ELISA method for detecting PEDV antibody was established, and when pregnant sows were immunized with PEDV inactivated vaccine before delivery, The maternal specific antibody positive rate and antibody level in serum of piglets were low and maintained for a short time.
【作者单位】：河北农业大学动物医学院河北省兽用生物制品工程技术研究中心;河北省畜牧兽医研究所;
【基金】：国家重点研发计划(2016YFD0500703) 国家自然科学基金(31372441)
【分类号】：S852.651

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