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IgG (IgG-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.%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 IgG (IgG-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 IgG-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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