【摘要】：為了解北京地區(qū)奶牛IBR流行情況,用IDEXX IBRgE抗體檢測試劑盒檢測來自北京不同地區(qū)的2582份牛血樣。結果顯示,調查的地區(qū)均有IBRV抗體陽性牛存在,其中昌平23.6%(245/1037),房山100%(50/50),延慶50.63%(202/399),順義60%(9/15),豐臺30.03%(164/546),密云43.46(232/535),總陽性率為34.93%(902/2582)。血清學調查結果表明,目前北京地區(qū)IBR流行情況嚴重,應盡快采取相應的措施。為建立一種檢測IBRV的方法,試驗將IBRV接種MDBK細胞,以濃縮純化的IBRV作為包被抗原,建立了間接ELISA檢測方法。通過棋盤法確定了抗原的最佳包被濃度和血清的最佳稀釋倍數(shù)�？乖粷舛�20μg/mL,4℃過夜;PBST洗滌;3%BSA的PBST作為封閉液,37℃封閉2h;血清用PBST作1:80稀釋,100μL/孔,37℃度作用1h;HRP-兔抗牛IgG用3%BSA的PBST作1:20000倍稀釋,37℃作用1h;TMB底物室溫避光顯色10min。陰、陽性血清臨界值為0.240。特異性試驗結果表明,IBRV陽性血清被檢為陽性,BVDV、牛白血病病毒、副結核病病毒陽性血清均被檢為陰性。敏感性試驗結果表明,IDEXX試劑盒中的標準陽性血清1:640倍稀釋時仍能檢出陽性抗體。與中和試驗相比,特異性為100%(23/23),敏感性為82.76%(24/29)。批內和批間重復性試驗的變異系數(shù)均小于10%,表明該方法重復性好。與IDEXX試劑盒相比,陽性符合率為87.7%(228/260),陰性符合率為98%(196/200)。以上結果表明所建立的間接ELISA方法可用于IBRV感染的監(jiān)測,并替代目前市場上昂貴的IBR進口檢測試劑盒。
[Abstract]:In order to understand the prevalence of IBR in dairy cows in Beijing, 2582 bovine blood samples from different areas of Beijing were detected with IDEXX IBRgE antibody test kit. The results showed that there were 23.6% (245r1037) in Changping, 100% (50 / 50) in Fangshan, 50.63% (2022 / 399) in Yanqing, 60% (9 / 15) in Shunyi, 30.03% (164 / 546) in Fengtai, 43.46 (232r535) in Miyun, the total positive rate was 34.93% (902 / 2582). The results of serological investigation showed that the epidemic situation of IBR in Beijing area was serious at present, and corresponding measures should be taken as soon as possible. In order to establish a method for detecting IBRV, an indirect Elisa method was established by inoculating IBRV into MDBK cells and using concentrated and purified IBRV as encapsulated antigen. The best coating concentration of antigen and the best dilution multiple of serum were determined by chessboard method. The concentration of antigenic coating was 20 渭 g / mL ~ (4 鈩，

本文編號：2119839