摘要
用胶体金标记提纯后的免抗鸡IgG,建立了一件以微孔滤膜为固相载体,以红色胶体金作为标记物的检测EDS-76病毒抗体的斑点免疫金测定法(DIGFA).该法具有简便、灵敏、快速等优点,全过程于3~5分钟内完成,阳性结果在膜上呈现红色斑点;对鸡IsG的最小检测量(灵敏度)为63×10-9g;与其它病毒阳性血清无交叉反应。DIGFA检测EDS-76抗体既可定性,也可定量。检测80份待检血清,在HI检出的56份阳性标本中(效价≥4log2)用DIGFA检出阳性标本55份(效价≥61og2)二者阳性符合率为98.2%.与HI比较,DIGFA平均高2.5(log2)。本技术为EDS-76和其它禽病的快速诊断和免疫监测提供了一个新的方法。
Abstract DIGPA with a microfilter paper as carrier and the red coiloidal gold as label was developed for detection of EDS-76 antibody in chicken sera by labelling the pruified rabbit anti-chick IgG with the prepared colloidal gold. The DIGPA had the advantages of simplity, high sensitivity. specificity and rapidity. The procedure was accomplished in 3-5 minutes. A positive result displayed a red dot on the paper. The minimum amount of purified chick IgG giving a positive reaction (1+. Reddot) was 63×10-9g/disc. The EDS-76 virus did not react with other viruses antisera by DIGFA. Antigen discsstored at room temperature were of normal reaction with EDS-76 anti-sera over a month. Detection for EDS-76 antibody by DIGFA combined the advantaged of qualitative and.quantitative analysis. 55 positive samples were detected by DIGFA(titer≥61og2) in the β-HI 56 positive sample(titer≥41og2).The positive coimcident rate was 98. 2 %. While compered with β-HI, the average titer of the DIGFA.was 2-3(log2) higher. The technique provided a new method for rapid disgnosis and immunodetection of EDS-76 and other avian diseases.