Italiya Jignesh, Straková Petra, Pavlačík Lukáš, Váhala Jiří, Hyjánek Jaroslav Haimy, Salát Jiří, Růžek Daniel, Komárková Dominika, Černý Jiří
Centre for Infectious Animal Diseases, Faculty of Tropical AgriSciences, Czech University of Life Sciences Prague, Kamýcká 129, CZ-16500 Prague-Suchdol, Czechia.
Veterinary Research Institute, Hudcova 70, CZ-62100 Brno, Czechia.
J Wildl Dis. 2025 Apr 1;61(2):396-407. doi: 10.7589/JWD-D-24-00028.
Coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), first emerged in Wuhan in 2019 and rapidly spread worldwide. During the course of the COVID-19 pandemic, numerous reports highlighted infections of wild animals by SARS-CoV-2. Nevertheless, further research is required to understand the virus potential to infect various animal species, which is crucial for evaluating its future evolution and the potential reemergence of SARS-CoV-2. The total concentration of immunoglobulin G (IgG) represents a valuable yet underused diagnostic parameter for health assessments in wild animals, primarily due to the absence of effective diagnostic tools. A protein A-based indirect ELISA can serve as an efficient method to identify IgG antibodies against different pathogens in wildlife surveillance programs. To develop a multispecies protein A-ELISA for IgG detection against SARS-CoV-2, we used 44 animal species serum samples to ascertain the protein A binding affinity, and 88 serum samples, chosen for the strong binding affinity to protein A, were used to identify IgG antibodies against SARS-CoV-2. The serum samples were obtained from animals housed in Safari Park Dvůr Králové, Czech Republic. The zoo animals are in proximity to humans, facilitating the exploration of potential reverse transmission events of SARS-CoV-2 from humans to animals. Also, they undergo routine veterinary examinations, providing convenient access to blood samples. Therefore, they can be easily used for the development of protein A-based indirect ELISA for wildlife disease surveillance programs. Based on the ELISA results, antibodies to SARS-CoV-2 were detected in the sera of 16 animals. To further confirm these findings, the ELISA-positive samples were subjected to virus neutralization assays. This additional testing revealed the presence of SARS-CoV-2 neutralizing antibodies in the serum of two white rhinoceros (Ceratotherium simum simum) and one Persian leopard (Panthera pardus tulliana).
2019冠状病毒病（COVID-19）由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起，于2019年首次在武汉出现并迅速在全球传播。在COVID-19大流行期间，大量报告强调了SARS-CoV-2对野生动物的感染情况。然而，仍需要进一步研究以了解该病毒感染各种动物物种的可能性，这对于评估其未来演变以及SARS-CoV-2可能的再次出现至关重要。免疫球蛋白G（IgG）的总浓度是野生动物健康评估中一个有价值但未得到充分利用的诊断参数，主要原因是缺乏有效的诊断工具。基于蛋白A的间接ELISA可作为一种有效的方法，用于在野生动物监测项目中鉴定针对不同病原体的IgG抗体。为了开发一种用于检测针对SARS-CoV-2的IgG的多物种蛋白A-ELISA，我们使用了44种动物的血清样本确定蛋白A的结合亲和力，并使用88份对蛋白A具有强结合亲和力的血清样本鉴定针对SARS-CoV-2的IgG抗体。血清样本取自捷克共和国德沃夏克拉洛韦野生动物园饲养的动物。这些动物园动物与人类接近，便于探索SARS-CoV-2从人类到动物的潜在反向传播事件。此外，它们接受常规兽医检查，便于获取血液样本。因此，它们可轻松用于开发基于蛋白A的间接ELISA，用于野生动物疾病监测项目。根据ELISA结果，在16只动物的血清中检测到了针对SARS-CoV-2的抗体。为了进一步证实这些发现，对ELISA阳性样本进行了病毒中和试验。这项额外检测揭示了两只白犀（Ceratotherium simum simum）和一只波斯豹（Panthera pardus tulliana）的血清中存在SARS-CoV-2中和抗体。
