本文選題：口蹄疫 + 黏膜免疫��； 參考：《中國(guó)畜牧獸醫(yī)》2017年06期

【摘要】：為建立一種檢測(cè)口蹄疫病毒(foot-and-mouth disease virus,FMDV)特異性IgA抗體的檢測(cè)方法,本研究以原核表達(dá)系統(tǒng)表達(dá)純化的FMDV結(jié)構(gòu)蛋白VP1作為包被抗原,以鼠抗豬IgA單克隆抗體為二抗,辣根過氧化酶標(biāo)記的羊抗鼠IgG抗體為三抗,建立豬A型FMDV特異性IgA抗體間接ELISA檢測(cè)方法。確定抗原包被濃度為3.50μg/mL,二抗與三抗的最佳稀釋度為1∶10 000,二抗和三抗作用時(shí)間均為30min。所建立的方法與抗豬瘟病毒、豬繁殖與呼吸道綜合征病毒等病原的特異性IgA抗體間無(wú)交叉反應(yīng),A型口蹄疫感染樣品的陽(yáng)性檢出率在90%以上,批內(nèi)和批間重復(fù)性試驗(yàn)的變異系數(shù)介于3.16%~9.76%。該方法為監(jiān)測(cè)FMDV特異性IgA抗體水平變化規(guī)律及豬的黏膜免疫效果評(píng)價(jià)及口蹄疫的早期診斷提供了一種新方法。
[Abstract]:In order to establish a method for the detection of foot-and-mouth disease virus (FMDV) specific IgA antibody, the purified FMDV structural protein VP1 was expressed in prokaryotic expression system as the coating antigen, and the mouse anti porcine IgA monoclonal antibody was used as the second antibody. The goat anti-mouse IgG antibody labeled by horseradish peroxidase (HRP) was a triple antibody. An indirect Elisa method for detection of swine FMDV specific IgA antibody was established. The concentration of antigen coating was 3.50 渭 g / mL, the best dilution of the second and third antibodies was 1:10, and the time of the second and third antibodies was 30 min. The positive rate of the method and the specific IgA antibody against swine fever virus, porcine reproductive and respiratory syndrome virus were over 90%. The coefficient of variation of intra-and inter-assay reproducibility tests ranged from 3.16 to 9.76. This method provides a new method for monitoring the changes of FMDV specific IgA antibody level, the evaluation of mucosal immune effect of pigs and the early diagnosis of foot-and-mouth disease.
【作者單位】： 中國(guó)農(nóng)業(yè)科學(xué)院蘭州獸醫(yī)研究所國(guó)家口蹄疫參考實(shí)驗(yàn)室家畜疫病病原生物學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室;
【基金】：“十三五”國(guó)家重點(diǎn)研發(fā)計(jì)劃(2016YFD0501503) 甘肅省科技支撐計(jì)劃(1604NKCA045-2) 國(guó)家生豬現(xiàn)代產(chǎn)業(yè)技術(shù)體系項(xiàng)目(CARS-36-06B)
【分類號(hào)】：S852.651

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