Development of a recombinant protein-based ELISA for detection of antibodies against bovine herpesvirus 6 (BoHV6).

IF 1.3 3区农林科学 Q2 VETERINARY SCIENCES

Journal of Veterinary Research Pub Date : 2023-12-19 eCollection Date: 2023-12-01 DOI:10.2478/jvetres-2023-0069

Piotr Kubiś, Jacek Kuźmak

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引用次数: 0

Abstract

Introduction: Bovine herpesvirus 6 (BoHV6) belongs to the Herpesviridae family, Gammaherpesvirinae subfamily and Macavirus genus. It is common in cattle, but was also detected in American bison (Bison bison) and water buffalo (Bubalus bubalis). The aim of the experiment was to develop an ELISA for serological examination of cattle sera for the presence of anti-BoHV6 specific antibodies.

Material and methods: Viral DNA from a BoHV6-positive cow was amplified by qPCR and the resulting fragments of the gB and gH genes encoding glycoproteins B and H (gB and gH) were cloned into the pLATE52 vector to express recombinant gB (rgB) and gH (rgH) in Rosetta (DE3) E. coli. The expressed recombinant proteins were used as antigens in the developed ELISA.

Results: The proteins expressed had the expected molecular weight. A total of 143 sera were examined, and 141 of them were positive, according to the chosen cut-off values of 9% and 10% for the sample-to-positive ratios of the rgB and rgH antigens, respectively.

Conclusion: The rgB and rgH recombinant antigens of BoHV6 were successfully expressed in E. coli and successfully used in a newly developed ELISA.

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开发基于重组蛋白的酶联免疫吸附试验，用于检测牛疱疹病毒 6 (BoHV6) 抗体。

导言：牛疱疹病毒 6（BoHV6）属于疱疹病毒科、γ疱疹病毒亚科和巨细胞病毒属。它常见于牛，但也在美洲野牛（Bison bison）和水牛（Bubalus bubalis）中检测到。实验的目的是开发一种酶联免疫吸附试验（ELISA），用于对牛血清进行血清学检查，以检测是否存在抗 BoHV6 的特异性抗体：通过 qPCR 扩增来自 BoHV6 阳性母牛的病毒 DNA，将得到的编码糖蛋白 B 和 H（gB 和 gH）的 gB 和 gH 基因片段克隆到 pLATE52 载体中，在 Rosetta（DE3）大肠杆菌中表达重组 gB（rgB）和 gH（rgH）。表达的重组蛋白在开发的酶联免疫吸附试验中用作抗原：结果：表达的蛋白质具有预期的分子量。共检测了 143 份血清，根据 rgB 和 rgH 抗原样本与阳性比值分别为 9% 和 10% 的临界值，其中 141 份血清呈阳性：结论：在大肠杆菌中成功表达了 BoHV6 的 rgB 和 rgH 重组抗原，并成功用于新开发的 ELISA。

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

Journal of Veterinary Research Veterinary-General Veterinary

CiteScore

0.90

自引率

5.60%

发文量

审稿时长

18 weeks

期刊介绍： Journal of Veterinary Research (formerly Bulletin of the Veterinary Institute in Pulawy) is a quarterly that publishes original papers, review articles and short communications on bacteriology, virology, parasitology, immunology, molecular biology, pathology, toxicology, pharmacology, and biochemistry. The main emphasis is, however, on infectious diseases of animals, food safety and public health, and clinical sciences.