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用于检测抗牛泡沫病毒抗体的基于重组蛋白的酶联免疫吸附测定法的开发

Development of a Recombinant Protein-based ELISA for Detection of Antibodies Against Bovine Foamy Virus.

作者信息

Materniak-Kornas Magdalena, Osiński Zbigniew, Rudzki Marcin, Kuźmak Jacek

机构信息

Department of Biochemistry, National Veterinary Research Institute, 24-100 Pulawy, Poland.

Departament of Hygiene of Feeding Stuffs, National Veterinary Research Institute, 24-100 Pulawy, Poland.

出版信息

J Vet Res. 2017 Sep 19;61(3):247-252. doi: 10.1515/jvetres-2017-0034. eCollection 2017 Sep.

Abstract

INTRODUCTION

Infections with bovine foamy virus (BFV) were found in many countries but there is a lack of large-scale surveys on the prevalence of BFV among dairy cattle. The aim of this study was to develop and validate the recombinant Gag protein-based ELISA and to estimate the prevalence of antibodies against BFV.

MATERIAL AND METHODS

Gag coding region from BFV was cloned into expression vector pT7Arg-STOP, which expressed a high level of recombinant Gag protein from . The ELISA was standardised, and the cut-off value and sensitivity and specificity of the test were calculated using a receiver operating characteristic and Bayesian estimation.

RESULTS

A total of 3,051 serum samples were tested by ELISA and 939 (30.8%) sera were recognised as positive. When Bayesian approach was used, the overall true BFV prevalence was 29.7% (95% CI: 25.9-33.4%).

CONCLUSION

Expressed Gag protein of BFV has been used successfully as an antigen for ELISA. Eventually, this study provides basic information about the epidemiological status of infection with BFV in dairy cattle in Poland, which can be used for further studies on dissemination and transmission of BFV infection.

摘要

引言

在许多国家都发现了牛泡沫病毒(BFV)感染,但缺乏关于奶牛中BFV流行情况的大规模调查。本研究的目的是开发并验证基于重组Gag蛋白的酶联免疫吸附测定(ELISA),并估计抗BFV抗体的流行率。

材料与方法

将BFV的Gag编码区克隆到表达载体pT7Arg-STOP中,该载体可高水平表达重组Gag蛋白。对ELISA进行标准化,并使用受试者工作特征曲线和贝叶斯估计计算该检测的临界值、敏感性和特异性。

结果

共通过ELISA检测了3051份血清样本,其中939份(30.8%)血清被判定为阳性。采用贝叶斯方法时,BFV的总体真实流行率为29.7%(95%置信区间:25.9 - 33.4%)。

结论

BFV表达的Gag蛋白已成功用作ELISA的抗原。最终,本研究提供了波兰奶牛中BFV感染流行病学状况的基本信息,可用于进一步研究BFV感染的传播和扩散。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8f05/5894420/f1b1bb8cb6b6/jvetres-61-247-g001.jpg

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