Rapid Detection of Getah Virus Antibodies in Horses Using a Recombinant E2 Protein-Based Immunochromatographic Strip

文献类型： 外文期刊

第一作者： Zhong, Dengke

作者： Zhong, Dengke;Zheng, Jiayang;Ma, Zhiyong;Wei, Jianchao;Wang, Yan

作者机构：

关键词： Getah virus; colloidal immunochromatographic strip; horse; rapid detection

期刊名称：ANIMALS （ 影响因子：2.7； 五年影响因子：3.2 ）

ISSN： 2076-2615

年卷期： 2024 年 14 卷 16 期

页码：

收录情况： SCI

摘要： Simple Summary Getah virus (GETV) is an important zoonotic pathogen with an extensive host range, including horses, pigs, cattle, sheep, birds, and humans, resulting in substantial losses in the livestock and agricultural industries; as such, the prevalence and impact of GETV are significant concerns in China. In this study, the antigen domain of the E2 glycoprotein of GETV was expressed and purified in the Escherichia coli expression system, and an on-site immunochromatographic strip (ICS) was successfully developed to detect GETV antibodies in horses. The ICS has the advantages of being rapid, sensitive, specific, and is an effective method for the detection of GETV in horses that does not rely on special equipment or skilled personnel and can be used for the field diagnosis of GETV in horses.Abstract The prevalence and impact of Getah virus (GETV) are significant concerns in China. GETV can infect a wide range of animals, including horses, pigs, sheep, cattle, birds, and humans, resulting in substantial losses in the livestock and agricultural industries. GETV infection can cause the development of ulcers and inflammation in the mouth and gums of horses, which result in pain and discomfort and lead to symptoms such as reduced appetite, drooling, and difficulty chewing. As a result, there is a pressing need for efficient and rapid disease diagnosis methods. However, the currently available diagnostic methods have limitations in terms of operational time, equipment, and the experience of the individuals using them. In this study, a rapid, specific, and sensitive detection method was developed using a colloidal gold-based immunochromatographic strip (ICS) for the detection of antibodies against GETV in horses. To prepare the ICS, the antigen domain of the E2 glycoprotein of GETV was expressed using the Escherichia coli expression system after analysis with DNAstar v7.1 software. The nitrocellulose membrane was coated with rE2 protein or SPA to form the test line and control line, respectively. After optimizing the reaction conditions, the sensitivity, specificity, and repeatability of the strip were verified. The results showed that the test strip had a detection limit of up to 1:320 dilutions for GETV-positive serum, with no cross-reactivity observed with other equine-susceptible pathogens such as equine arteritis virus (EAV), equine herpesvirus-1 (EHV-I), equine infectious anemia virus (EIAV), equine influenza virus (EIV), African horse sickness virus (AHSV), and Japanese encephalitis virus (JEV). Furthermore, the ICS exhibited a concordance rate of 94.0% when testing 182 clinical serum samples compared to the virus neutralization test. Overall, this ICS diagnosis method will be an effective tool for the rapid detection of GETV in the field.

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