雙抗體夾心ELISA檢測小反芻獸疫病毒抗原方法的建立及病原流行病學調(diào)查
發(fā)布時間:2018-07-07 20:22
本文選題:PPRV + 小反芻獸疫 ; 參考:《中國獸醫(yī)學報》2017年05期
【摘要】:應(yīng)用表達純化的小反芻獸疫病毒(peste des petits ruminants virus,PPRV)N蛋白制備的多克隆抗體,建立了檢測PPRV抗原的雙抗體夾心ELISA方法,并對吉林省和內(nèi)蒙古地區(qū)羊群感染PPRV進行了調(diào)查。方陣法確定了抗PPRV兔源IgG作為捕獲抗體的包被量為0.2μg,酶標抗體的最佳稀釋度為1∶1 000。對大量小反芻獸疫陰性糞便樣品進行檢測及統(tǒng)計學處理,確定了雙抗體夾心ELISA檢測PPRV的判定標準,即被檢糞便樣品D490≥0.221,判定為陽性。特異性、敏感性等試驗結(jié)果表明,建立的檢測PPRV抗原方法具有特異、敏感和快速等優(yōu)點。與RT-PCR方法相比,該方法省時省力、簡單快速。應(yīng)用建立的檢測PPRV抗原的雙抗體夾心ELISA對吉林省和內(nèi)蒙古不同地區(qū)的羊糞樣進行檢測,發(fā)現(xiàn)羊群均存在程度不同的PPRV隱性感染。本研究在國內(nèi)首次揭示出臨床健康羊群攜帶PPRV,為今后小反芻獸疫的診斷與防控提供了新的流行病學理論依據(jù)。
[Abstract]:A double antibody sandwich Elisa method was developed to detect (peste des petits ruminants virus antigen by using polyclonal antibody prepared from purified protein of small ruminant virus (peste des petits ruminants virus) N. The infection of sheep in Jilin province and Inner Mongolia was investigated. The anti-PPRV rabbit IgG was determined by square matrix method as the encapsulation amount of the capture antibody was 0.2 渭 g, and the best dilution of the enzyme labeled antibody was 1:1 000. A large number of small ruminant negative fecal samples were detected and statistically analyzed, and the standard of double antibody sandwich Elisa for PPRV detection was determined, that is, D490 鈮,
本文編號:2106162
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