猪传染性胃肠炎病毒间接免疫荧光检测方法的建立

发布时间：2018-03-28 22:13

  本文选题：猪传染性胃肠炎病毒　切入点：间接免疫荧光方法　出处：《中国兽医科学》2017年01期

【摘要】：为建立猪传染性胃肠炎病毒(TGEV)直观、特异和快速的检测方法,将TGEV接种PK-15细胞后,以抗TGEV N蛋白的单克隆抗体为一抗,荧光素FITC标记的山羊抗小鼠Ig G为二抗,建立了TGEV的间接免疫荧光(IFA)检测方法。结果显示,TGEV的最佳接种效价为1×102TCID50,培养时间为24 h,细胞最佳固定液为40 m L/L多聚甲醛,最佳封闭条件为10 g/L BSA 37℃封闭30 min,一抗的最佳使用浓度为1 ng/L,二抗的最佳稀释度为1∶100。对常见的猪流行性腹泻病毒、猪轮状病毒和猪圆环病毒2型等猪病病原检测均为阴性。结果表明,建立的检测方法对TGEV具有良好的特异性。本研究建立的方法为TGEV的实验室诊断及TGEV在感染细胞中的定位和动态分布提供了有效的检测手段。
[Abstract]:In order to establish an intuitive, specific and rapid method for detection of porcine infectious gastroenteritis virus, TGEV was inoculated into PK-15 cells. Monoclonal antibody against TGEV N protein was used as the first antibody and goat anti-mouse IgG labeled with fluorescein FITC as the second antibody. An indirect immunofluorescence assay was established for the detection of TGEV. The results showed that the best titer of TGEV was 1 脳 102 TCID 50, the culture time was 24 h, and the best immobilization solution was 40 mL / L polyformaldehyde. The best sealing condition is 10 g / L BSA 37 鈩，

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