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檢測(cè)豬流行性腹瀉病毒S1蛋白抗體的間接ELISA方法

發(fā)布時(shí)間:2018-03-07 11:46

  本文選題:豬流行性腹瀉病毒 切入點(diǎn):重組PEDV 出處:《畜牧獸醫(yī)學(xué)報(bào)》2017年06期  論文類型:期刊論文


【摘要】:為了建立檢測(cè)豬流行性腹瀉病毒(porcine epidemic diarrhea virus,PEDV)抗體的間接ELISA方法,初步分析PED滅活疫苗免疫母豬所產(chǎn)仔豬的血清母源特異抗體動(dòng)態(tài),筆者應(yīng)用重組PEDV部分纖突(spike,S)蛋白作為檢測(cè)抗原,建立了基于PEDV S1蛋白的間接ELISA抗體檢測(cè)方法,并用該方法檢測(cè)了產(chǎn)前4周用PED滅活疫苗免疫母豬的初乳、分娩當(dāng)天血清及其仔豬1、7、14、21、28和35日齡(d)時(shí)血清中的PEDV抗體。結(jié)果顯示:建立的間接ELISA方法能夠特異地檢測(cè)PEDV抗體,與豬瘟病毒、豬圓環(huán)病毒2型、豬繁殖與呼吸綜合征病毒、偽狂犬病病毒、豬傳染性胃腸炎病毒及豬輪狀病毒的抗血清無交叉反應(yīng);敏感性高于免疫過氧化物酶單層試驗(yàn)(immunoperoxidase monolayer assay,IPMA),與IPMA檢測(cè)結(jié)果的符合率為97.14%;批內(nèi)和批間重復(fù)性試驗(yàn)的變異系數(shù)小于10%。免疫母豬初乳、分娩當(dāng)天血清中的PEDV抗體陽(yáng)性率為100%,初乳比血清中的平均抗體水平低;1d仔豬血清PEDV抗體陽(yáng)性率為50%,平均抗體水平低,與初乳中的抗體水平接近,7d以后所有仔豬血清全部轉(zhuǎn)為PEDV抗體陰性。建立了檢測(cè)PEDV抗體的間接ELISA方法;產(chǎn)前用PEDV滅活疫苗給懷孕母豬免疫1次時(shí),其所產(chǎn)仔豬血清中的母源特異抗體陽(yáng)性率和抗體水平低,且維持時(shí)間短暫。
[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.
【作者單位】: 河北農(nóng)業(yè)大學(xué)動(dòng)物醫(yī)學(xué)院河北省獸用生物制品工程技術(shù)研究中心;河北省畜牧獸醫(yī)研究所;
【基金】:國(guó)家重點(diǎn)研發(fā)計(jì)劃(2016YFD0500703) 國(guó)家自然科學(xué)基金(31372441)
【分類號(hào)】:S852.651

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