[Virus-like particles based on rotavarus A recombinant VP2/VP6 proteins for assessment the antibody immune response by ELISA].

Q3 Medicine Voprosy virusologii Pub Date : 2023-05-18 DOI:10.36233/0507-4088-169

I E Filatov, V V Tsibezov, M V Balandina, S N Norkina, O E Latyshev, O V Eliseeva, S A Cherepushkin, O A Verkhovsky, T V Grebennikova

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引用次数: 1

Abstract

Introduction: Rotavirus infection is one of the main concerns in infectious pathology in humans, mammals and birds. Newborn piglets or rodents are usually being used as a laboratory model for the evaluation of immunogenicity and efficacy for all types of vaccines against rotavirus A (RVA), and the use of ELISA for the detection of virus-specific antibodies of specific isotype is an essential step of this evaluation.

Objective: Development of indirect solid-phase ELISA with VP2/VP6 rotavirus VLP as an antigen to detect and assess the distribution of RVA-specific IgG, IgM and IgA in the immune response to rotavirus A.

Materials and methods: VP2/VP6 rotavirus VLP production and purification, electron microscopy, PAGE, immunoblotting, ELISA, virus neutralization assay.

Results: The study presents the results of development of a recombinant baculovirus with RVA genes VP2-eGFP/VP6, assessment of its infectious activity and using it for VLP production. The morphology of the VP2/VP6 rotavirus VLPs was assessed, the structural composition was determined, and the high antigenic activity of the VLP was established. VLP-based ELISA assay was developed and here we report results for RVA-specific antibody detection in sera of different animals.

Conclusion: The developed ELISA based on VP2/VP6 rotavirus VLP as a universal antigen makes it possible to detect separately IgG, IgM and IgA antibodies to rotavirus A, outlining its scientific and practical importance for the evaluation of immunogenicity and efficacy of traditional vaccines against rotavirus A and those under development.

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[基于轮状虫A重组VP2/VP6蛋白的病毒样颗粒ELISA检测抗体免疫应答]。

导论:轮状病毒感染是人类、哺乳动物和鸟类感染病理学的主要问题之一。新生仔猪或啮齿动物通常被用作实验室模型，用于评估所有类型的轮状病毒a (RVA)疫苗的免疫原性和有效性，而使用ELISA检测特定同型的病毒特异性抗体是这一评估的重要步骤。目的:建立以VP2/VP6轮状病毒VLP为抗原的间接固相ELISA法，检测和评价rva特异性IgG、IgM和IgA在轮状病毒a免疫应答中的分布。材料和方法:VP2/VP6轮状病毒VLP的制备和纯化、电镜、PAGE、免疫印迹、ELISA、病毒中和试验。结果:本研究获得了RVA基因VP2-eGFP/VP6重组杆状病毒，并对其感染活性进行了评价，并将其用于VLP的生产。鉴定了VP2/VP6轮状病毒VLP的形态，确定了VLP的结构组成，证实了VLP具有较高的抗原性。我们建立了基于vlp的ELISA检测方法，报告了不同动物血清中rva特异性抗体的检测结果。结论:所建立的以VP2/VP6轮状病毒VLP为通用抗原的酶联免疫吸附测定法能够分别检测轮状病毒a的IgG、IgM和IgA抗体，对评价传统和正在开发的轮状病毒a疫苗的免疫原性和疗效具有重要的科学意义和实用价值。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Voprosy virusologii Medicine-Infectious Diseases

CiteScore

2.00

自引率

0.00%

发文量

期刊介绍： The journal deals with advances in virology in Russia and abroad. It publishes papers dealing with investigations of viral diseases of man, animals and plants, the results of experimental research on different problems of general and special virology. The journal publishes materials are which promote introduction into practice of the achievements of the virological science in the eradication and incidence reduction of infectious diseases, as well as their diagnosis, treatment and prevention. The reader will find a description of new methods of investigation, new apparatus and devices.