本文關(guān)鍵詞：基于CdtB的副豬嗜血桿菌病間接ELISA檢測方法的建立　出處：《黑龍江八一農(nóng)墾大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 副豬嗜血桿菌 rCdtB 間接ELISA

【摘要】：為建立一種快速檢測副豬嗜血桿菌的血清學(xué)方法,本實驗采用重組表達(dá)的H.parasuis細(xì)胞致死膨脹毒素B亞基蛋白(rCdtB)作為包被抗原,并對各反應(yīng)條件進(jìn)行優(yōu)化,建立了檢測H.parasuis血清抗體的間接ELISA方法。以該方法檢測豬傳染性胃腸炎病毒、豬偽狂犬病病毒、圓環(huán)病毒2型、豬瘟病毒、豬病毒性腹瀉病毒、豬藍(lán)耳病病毒、大腸桿菌、豬鏈球菌2型、多殺性巴氏桿菌、胸膜肺炎放線桿菌和H.parasuis陽性血清,除了H.parasuis陽性血清呈陽性外,其余均為陰性,表明該方法特異性良好。批內(nèi)重復(fù)和批間重復(fù)的變異系數(shù)分別為小于7%和8%。此外,將rCdt B-ELISA方法與商品化試劑盒BioChek HPS-OppA間接ELISA抗體檢測方法對背景明確的116份H.parasuis陽性血清和112份H.parasuis陰性血清進(jìn)行檢測,商品化試劑盒陽性符合率為25%,陰性血清符合率為100%,總符合率為62%,其中rCdtB-ELISA方法對陽性血清的檢出率高于商品化試劑盒。應(yīng)用rCdt B-ELISA方法檢測330份臨床豬血清樣品,結(jié)果顯示,副豬嗜血桿菌抗體陽性檢出率為11.8%。應(yīng)用該方法對H.parasuis 7份陽性血清及4份陰性血清進(jìn)行保質(zhì)期試驗,結(jié)果發(fā)現(xiàn)6個月后S/P值均大于0.2417,陰性血清的S/P值均小于0.2417,證明rCdtB-ELISA試劑盒至少可以保存6個月。
[Abstract]:In order to establish a rapid serological method for the detection of Haemophilus parapleus, the recombinant H. parasuis cell line lethal expansion toxin B subunit protein (rCdtB) was used as the coating antigen. The indirect ELISA method for detection of H. parasuis serum antibody was established. The method was used to detect porcine infectious gastroenteritis virus and porcine pseudorabies virus. Circovirus type 2, swine fever virus, porcine viral diarrhea virus, porcine blue ear disease virus, Escherichia coli, Streptococcus suis type 2, Pasteurella multocida. The positive sera of Actinobacillus pleuropneumoniae and H. parasuis were negative except H. parasuis positive serum. The results showed that the specificity of the method was good. The coefficients of variation of intra-and inter-assay repeats were less than 7% and 8, respectively. RCdt B-ELISA and Commercial Kit BioChek. HPS-OppA indirect ELISA antibody assay was used to detect 116 H. parasuis positive sera and 112 H. parasuis negative sera. The positive coincidence rate of commercial kit and negative serum were 25 and 100 respectively, and the total coincidence rate was 62%. The detection rate of positive serum by rCdtB-ELISA method was higher than that by commercial kit. RCdtB-ELISA method was used to detect 330 clinical pig serum samples. The positive rate of Haemophilus parasuis antibody was 11.8%. This method was used to test the shelf life of 7 positive sera and 4 negative sera of H. parasuis. The results showed that after 6 months, the values of S / P were more than 0.2417, and those of negative serum were less than 0.2417. It was proved that the rCdtB-ELISA kit could be kept for at least 6 months.
【學(xué)位授予單位】：黑龍江八一農(nóng)墾大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S855.12

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