Tarasiuk Karolina, Holec-Gąsior Lucyna, Ferra Bartłomiej, Rapak Andrzej
Department of Poultry Diseases, National Veterinary Research Institute, Partyzantów 57 Avenue, 24-100, Puławy, Poland.
Department of Microbiology, Gdańsk University of Technology, Gabriela Narutowicza 11/12, 80-233, Gdańsk, Poland.
BMC Vet Res. 2019 Aug 1;15(1):274. doi: 10.1186/s12917-019-2027-1.
In Poland, the leader in goose production in Europe, goose parovirus infection, or Derzsy's disease (DD), must be reported to the veterinary administration due to the serious economic and epizootic threat to waterfowl production. Prophylactic treatment for DD includes attenuated live or inactivated vaccines. Moreover, the control of DD includes the monitoring of maternal derived antibody (MDA) levels in the offspring and antibody titers in the parent flock after vaccination. The aim of this study was to develop an ELISA for the detection of goose parvovirus (GPV) antibodies.
Two recombinant protein fragments derived from VP3 (viral protein 3) GPV, namely VP3ep6 and VP3ep4-6 with a mass of 20.9 and 32.3 kDa, respectively, were produced using an Escherichia coli expression system. These proteins were purified by one-step nickel-affinity chromatography, which yielded protein preparations with a purity above 95%. These recombinant proteins were useful in the detection of serum anti-GPV antibodies, and this was confirmed by Western blotting. However, recombinant VP3ep4-6 protein showed a greater ability to correctly identify sera from infected geese. In the next stage of the project, a pool of 166 goose sera samples, previously examined by a virus neutralization test (VN), was tested. For further studies, one recombinant protein (VP3ep4-6) was selected for optimization of the test conditions. After optimization, the newly developed ELISA was compared to other serological tests, and demonstrated high sensitivity and specificity.
In conclusion, the VP3ep4-6 ELISA method described here can be used for the detection of antibodies to GPV in serum.
在欧洲鹅生产领域处于领先地位的波兰，鹅细小病毒感染，即德尔泽氏病（DD），因其对水禽生产构成严重经济和 epizootic 威胁，必须向兽医管理部门报告。DD 的预防性治疗包括减毒活疫苗或灭活疫苗。此外，DD 的防控包括监测后代中母源抗体（MDA）水平以及接种疫苗后亲代群体中的抗体滴度。本研究的目的是开发一种用于检测鹅细小病毒（GPV）抗体的酶联免疫吸附测定（ELISA）。
使用大肠杆菌表达系统制备了两个源自 GPV 的 VP3（病毒蛋白 3）的重组蛋白片段，即 VP3ep6 和 VP3ep4 - 6，质量分别为 20.9 和 32.3 kDa。这些蛋白通过一步镍亲和层析进行纯化，得到纯度高于 95%的蛋白制剂。这些重组蛋白可用于检测血清抗 GPV 抗体，蛋白质印迹法证实了这一点。然而，重组 VP3ep4 - 6 蛋白在正确识别感染鹅血清方面表现出更强的能力。在项目的下一阶段，对一组先前通过病毒中和试验（VN）检测的 166 份鹅血清样本进行了测试。为进一步研究，选择了一种重组蛋白（VP3ep4 - 6）来优化测试条件。优化后，将新开发的 ELISA 与其他血清学检测方法进行比较，结果显示其具有高灵敏度和特异性。
总之，本文所述的 VP3ep4 - 6 ELISA 方法可用于检测血清中 GPV 的抗体。
