禽白血病A亚群病毒gp85的单因子血清制备及其特异性鉴定

Mono-specific serum preparation and specificity of gp85 gene of subgroup A Avian Leukosis Virus

摘要:

摘要：【目的】为了研究出一种能够针对A亚群禽白血病的快速特异性诊断试剂。【方法】将A亚群禽白血病病毒（ALV-A）SDAU09E1株接种于DF1细胞上，以感染细胞DNA为模板，通过PCR方法扩增出1023bp的ALV-A-gp85基因。将其正确阅读框架插入表达载体PET-32a（+）中，实现在BL21（Rosetta）宿主菌中表达。将纯化的融合蛋白常规免疫小鼠，制备得抗血清。【结果】实验成功获得52.8kDa的重组融合蛋白，且具有良好的免疫原性。间接免疫荧光试验（IFA）表明该血清可与ALV-A和ALV-

Abstract:

Abstract: 【Objective】 In order to get a rapid specific diagnostic reagent for subgroup A Avian Leukosis Virus detection.【Methods】The Avian Leukosis Virus Subgroup A(ALV-A) SDAU09E1 strain was inoculated into DF1 cells, an ALV-A-gp85 DNA fragment of 1023bp was amplified from infected cells and inserted into PET-32a(+) plasmid at the location between restriction endonucleases BamHⅠand NotⅠsites. The recombinant plasmid PET-SDAU09E1-gp85 was transformed into E coli.BL21 (Rosetta) for gp85 gene expression. Then we used the purified recombinant fusion protein to immunize 6 weeks old Kunming white mice, and the antiserum were prepared. 【Results】The recombinant ALV-A gp85 fusion protein with a molecular weight of 52.8kDa demonstrated a good antigenecity. Mon-specific serum produced by vaccinated mice came out reactive with subgroups A and B ALV(ALV-A and ALV-B but not subgroup J ALV) by the indirect immunofluorescence (IFA) method. 【Conclusion】This was the first time to demonstrate a mono-specific antiserum specific to ALV-A and ALV-B,it could be used for differential diagnosis of exogenous ALV infections in CEF cultures when in complement with ALV-J specific monoclonal antibodies. Chickens in our country are now distressed by both classic ALV-A/B and emerging ALV-J, making differential diagnosis necessary, so studying this reagent has high practical value.