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使用基于重组E2蛋白的免疫层析试纸条快速检测马中的盖塔病毒抗体

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

作者信息

Zhong Dengke, Zheng Jiayang, Ma Zhiyong, Wang Yan, Wei Jianchao

机构信息

Department of Animal Science and Technology, Shanghai Vocational College of Agriculture and Forestry, Shanghai 201699, China.

Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China.

出版信息

Animals (Basel). 2024 Aug 8;14(16):2309. doi: 10.3390/ani14162309.

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 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.

摘要

在中国,盖塔病毒(GETV)的流行情况及其影响是备受关注的问题。GETV可感染多种动物,包括马、猪、羊、牛、鸟类和人类,给畜牧业和农业产业造成重大损失。GETV感染可导致马的口腔和牙龈出现溃疡和炎症,引发疼痛和不适,并导致食欲减退、流口水和咀嚼困难等症状。因此,迫切需要高效快速的疾病诊断方法。然而,目前可用的诊断方法在操作时间、设备以及使用人员的经验方面存在局限性。在本研究中,开发了一种基于胶体金免疫层析试纸条(ICS)的快速、特异且灵敏的检测方法,用于检测马血清中抗GETV的抗体。为制备ICS,使用DNAstar v7.1软件分析后,利用表达系统表达GETV E2糖蛋白的抗原结构域。将重组E2蛋白或SPA分别包被在硝酸纤维素膜上,形成检测线和对照线。优化反应条件后,对试纸条的灵敏度、特异性和重复性进行了验证。结果表明,该试纸条对GETV阳性血清的检测限高达1:320稀释度,与马动脉炎病毒(EAV)、马疱疹病毒1型(EHV-I)、马传染性贫血病毒(EIAV)、马流感病毒(EIV)、非洲马瘟病毒(AHSV)和日本脑炎病毒(JEV)等其他马易感病原体无交叉反应。此外,与病毒中和试验相比,ICS检测182份临床血清样本时的符合率为94.0%。总体而言,这种ICS诊断方法将成为现场快速检测GETV的有效工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0460/11350704/4d95fc7d0b3d/animals-14-02309-g001.jpg

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