基因C型鸭甲肝病毒间接免疫荧光检测方法的建立与初步应用

发布时间：2018-06-12 12:43

  本文选题：基因C型鸭甲肝病毒 + 间接免疫荧光技术　； 参考：《中国预防兽医学报》2017年04期

【摘要】：为建立基因C型鸭甲肝病毒(DHAV-C)间接免疫荧光检测技术(IFA),本研究以纯化的DHAV-C单克隆抗体(MAb)为一抗,FITC标记的兔抗鼠Ig G(Ig G-FITC)为二抗,建立了DHAV-C IFA检测方法。该方法仅对感染DHAV-C的肝组织触片检测为阳性,而对分别感染基因A型鸭甲肝病毒、新城疫病毒、鸭瘟病毒、鸭圆环病毒、鸭源金黄色葡萄球菌、鸭源大肠杆菌、鸭源多杀性巴氏杆菌、鸭源沙门氏菌的SPF鸭肝组织触片检测为阴性,表明该方法特异性好;重复性试验表明,该方法批内和批间重复性好。一抗和二抗在-20℃至少能保存7个月。实验感染SPF雏鸭IFA和RT-PCR检测符合率100%。本研究建立的DHAV-C IFA方法特异性好,可以用于该病毒感染肝组织的快速检测。
[Abstract]:In order to establish an indirect immunofluorescence assay for detection of DHAV-C gene type C duck hepatitis A virus (DHAV-C), a new method for detection of DHAV-C IFA was developed, using purified DHAV-C monoclonal antibody (MAb) as a second antibody against rabbit anti-FITC labeled rabbit anti-mouse Ig G G FITC. The method was only positive for DHAV-C infected liver tissue, but positive for duck hepatitis A virus, Newcastle disease virus, duck plague virus, duck circovirus, duck Staphylococcus aureus, duck Escherichia coli. The detection of Pasteurella multocida from duck and salmonella from duck was negative, which indicated that the method was specific and reproducible, and the repeatability test showed that the method had good reproducibility within and between batches. The first and second antibodies can be preserved at-20 鈩，

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