發(fā)布時間：2018-11-12 13:46

【摘要】：【目的】布魯氏菌病(Brucellosis)簡稱布病,是由布魯氏菌引起的以感染家畜為主的人畜共患傳染病,造成嚴(yán)重的公共衛(wèi)生問題。目前全世界范圍內(nèi)消除該病的主要方法是撲殺與免疫相結(jié)合,所以建立快速準(zhǔn)確的診斷方法對防治和清除布病非常必要。本文建立布魯氏菌病熒光偏振(FPA)抗體檢測方法,為布魯氏菌病(布病)的快速高效診斷提供技術(shù)手段。【方法】提純豬種布魯氏菌S2株脂多糖O鏈(OPS),經(jīng)異硫氰酸熒光素(FITC)標(biāo)記后作為診斷抗原。通過對樣品稀釋液、抗原稀釋度、反應(yīng)時間等條件的優(yōu)化,初步建立了布魯氏菌熒光偏振診斷方法。用該方法對148份布病陽性血清(其中牛血清70份,羊血清78份)和155份布病陰性血清(其中牛血清82份,羊血清73份)進(jìn)行檢測,確定其敏感性和特異性。按確定的技術(shù)參數(shù),制備3批布魯氏菌FPA抗體檢測試劑盒,使用質(zhì)控陰、陽性血清分別評價試劑盒的批內(nèi)和批間重復(fù)性。用400份臨床樣本比較本研究開發(fā)試劑盒與商品化進(jìn)口FPA試劑盒的符合率。【結(jié)果】使用0.5%蔗糖磷酸緩沖液作為血清樣品稀釋液;標(biāo)記抗原的使用濃度為90μg/m L;最佳反應(yīng)時間為3-5 min。本檢測方法的判定標(biāo)準(zhǔn)為:δm P值20時為陰性,δm P值≥20時為陽性。按上述條件建立的FPA檢測148份布病陽性血清和155份布病陰性血清,結(jié)果敏感性為98.6%,特異性為98.7%。對400份臨床樣本的比對檢測顯示,研究建立的FPA方法與進(jìn)口商品化試劑盒的總符合率為94.0%�！窘Y(jié)論】研究建立的布魯氏菌PFA抗體檢測方法具有良好的特異性和敏感性,可作為一種重要的布病診斷快速診斷方法。
[Abstract]:[objective] brucellosis (Brucellosis) is a zoonotic infectious disease caused by brucellosis, which causes serious public health problems. At present, the main method to eliminate the disease worldwide is the combination of culling and immunity, so it is necessary to establish a rapid and accurate diagnostic method for preventing and eliminating brucellosis. In this paper, a fluorescence polarization (FPA) antibody detection method for brucellosis was established, which provides a technical means for rapid and efficient diagnosis of brucellosis (brucellosis). [methods] purification of lipopolysaccharide O chain (OPS), from Brucella spp S2 strain Fluorescein isothiocyanate (FITC) was used as diagnostic antigen. Based on the optimization of sample dilution, antigen dilution and reaction time, a fluorescence polarization diagnostic method for brucella was established. This method was used to detect 148 brucellosis positive serum (70 bovine serum, 78 sheep serum) and 155 brucellosis negative serum (82 bovine serum and 73 sheep serum). The sensitivity and specificity of the method were determined. According to the determined technical parameters, three batches of brucella FPA antibody detection kits were prepared, and the intra- and inter-batch repeatability of the kits were evaluated by using quality control negative and positive serum respectively. The coincidence rate between the developed kit and the commercial FPA kit was compared with 400 clinical samples. [results] 0.5% sucrose phosphate buffer solution was used as the diluent of serum sample, and the concentration of labeled antigen was 90 渭 g / mL. The optimum reaction time is 3-5 min.. The criterion of this method is that 未 MP is negative at 20:00 and 未 MP 鈮，

本文編號：2327286