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当前位置: 首页 > 文章 > 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 中国预防兽医学报 2017 (4) 325-328
Position: Home > Articles > Establishment of indirect immunofluorescent assay of duck hepatitis A virus genotype C Chinese Journal of Preventive Veterinary Medicine 2017 (4) 325-328

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

作  者：
张焕容;何美琳;徐凤;岳华
单  位：
西南民族大学生命科学与技术学院
关键词：
基因C型鸭甲肝病毒;间接免疫荧光技术
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。
译  名：
Establishment of indirect immunofluorescent assay of duck hepatitis A virus genotype C
作  者：
ZHANG Huan-rong;HE Mei-lin;XU Feng;YUE Hua;College of Life Science and Technology,Southwest University for Nationalities;
关键词：
duck hepatitis A virus genotype C;;indirect inmmunofluorecent assay
摘  要：
The objective of this study is to establish the indirect immunofluorescent assay(IFA)to detect duck hepatitis A virus genotype C(DHAV-C). The indirect IFA was developed with the purified DHAV-C MAb as the first antibody and the commercial FITC labeled rabbit anti-mouse Ig G(Ig G-FITC) as the second antibody. Under the opitimized conditions for the assay, the specific test showed that when the liver tissue wafer sections prepared from DHAV-C, DHAV-A, NDV, DPV, DUCV, duck Staphylococcus aureus, duck Escherichia coli, duck Pasteurella and duck Salmonella respectively infected SPF ducklings were detected with the IFA, the specific fluorescences were seen only on DHAV-C infected tissue sections. The DHAV-C IFA showed good intra-and inner-batch repeatability. The first antibody DHAV-C MAb and the secondary Ig G-FITC was valid for at least 7 months stored at -20 ℃. The experimental infected SPF duckling livers were detected with both IFA and RT-PCR with 100% coincidence rate. This method can be used to detect DHAV-C infected liver tissue rapidly.

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