Development and evaluation of a gp85 protein-based subgroup-specific indirect enzyme-linked immunosorbent assay for the detection of anti-subgroup J avian leukosis virus antibodies
Development and evaluation of a gp85 protein-based subgroup-specific indirect enzyme-linked immunosorbent assay for the detection of anti-subgroup J avian leukosis virus antibodies
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开发和评估基于 gp85 蛋白的亚组特异性间接酶联免疫吸附测定法，用于检测抗亚组 J 禽白血病病毒抗体
DOI：
10.1007/s00253-019-10320-4
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发表时间：
2020-02-01
影响因子：
5
通讯作者：
Gao, Yulong
中科院分区：
文献类型：
--
作者：
Chang, Fangfang;Xing, Lixiao;Gao, Yulong
J亚群禽白血病病毒（ALV-J）是一种重要的禽类肿瘤病原，给养禽业造成重大的经济损失。血清学检测抗ALV-J感染的特异性抗体对临床成功诊断具有重要意义。在此，建立了稳定表达gp 85蛋白的293 F稳定细胞系。在该细胞系中，gp 85蛋白的表达量约为30 mg/L。以ALV-J gp 85蛋白为包被抗原，建立了检测ALV-J抗体的亚群特异性间接酶联免疫吸附试验（iELISA），其灵敏度（血清稀释度1：51200）是间接免疫荧光试验（IFA;血清稀释度1：3200）的16倍。此外，与其他常见禽病毒和其他禽白血病病毒亚群（如A和B亚群）的抗体无交叉反应性。通过实验感染和临床标本进一步验证了该方法的实用性。实验感染结果表明，免疫ELISA法在感染后4周即可检测到抗ALV-J抗体，至感染后20周抗体仍呈阳性，抗体阳性率为11.1%~ 33.3%。对临床标本的检测结果表明，抗ALV-J抗体阳性率为9.49%，与IFA的符合率为99.24%。总体而言，这些结果表明，在本研究中开发的亚组特异性iELISA具有良好的灵敏度，特异性和可行性。该方法可用于鸡场ALV-J的流行病学监测、诊断和根除。
Avian leukosis virus subgroup J (ALV-J) is an important pathogen for various neoplasms and causes significant economic losses in the poultry industry. Serological detection of specific antibodies against ALV-J infection is important for successful clinical diagnosis. Here, a 293F stable cell line was established to stably express gp85 protein. In this cell line, gp85 protein was expressed at approximately 30 mg/L. A subgroup-specific indirect enzyme-linked immunosorbent assay (iELISA) was developed using ALV-J gp85 protein as coated antigen to detect antibodies against ALV-J. The sensitivity of the iELISA (1:51200 diluted in serum) was 16 times more than that of indirect immunofluorescence assay (IFA; 1:3200 diluted in serum). Moreover, there was no crossreactivity with antibodies against other common avian viruses and other avian leukosis virus subgroups, such as subgroups A and B. The practicality of the iELISA was further evaluated by experimental infection and clinical samples. The results from experimental infection indicated that anti-ALV-J antibodies were readily detected by iELISA as early as 4 weeks after ALV-J infection, and positive antibodies were detected until 20 weeks, with an antibody-positive rate of 11.1% to 33.3%. Moreover, analysis of clinical samples showed that 9.49% of samples were positive for anti-ALV-J antibodies, and the concordance rate of iELISA and IFA was 99.24%. Overall, these results suggested that the subgroup-specific iELISA developed in this study had good sensitivity, specificity, and feasibility. This iELISA will be very useful for epidemiological surveillance, diagnosis, and eradication of ALV-J in poultry farms.