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寨卡病毒非结构蛋白1:异源表达、纯化及用于寨卡病毒感染诊断的潜力

Non-structural protein 1 from Zika virus: Heterologous expression, purification, and potential for diagnosis of Zika infections.

作者信息

Magalhães Ilana C L, Marques Lívia E C, Souza Pedro F N, Girão Nicolas M, Herazo Maestre M A, Costa Helen P S, vanTilburg Maurício F, Florean Eridan O P T, Dutra Rosa F, Guedes Maria Izabel F

机构信息

Biotechnology and Molecular Biology Laboratory, State University of Ceara, Fortaleza, Ceara, Brazil.

Biotechnology and Molecular Biology Laboratory, State University of Ceara, Fortaleza, Ceara, Brazil.

出版信息

Int J Biol Macromol. 2021 Sep 1;186:984-993. doi: 10.1016/j.ijbiomac.2021.07.083. Epub 2021 Jul 19.

Abstract

Zika virus (ZIKV) infections were associated with neurological disorders only after the Brazilian outbreak in 2015. The lack of vaccines and precise diagnosis requires a precise method to detect ZIKV infection. This study aimed to evaluate three ZIKV recombinant proteins for the development of ZIKV infections. Here, it was purified stable recombinant ZIKV Capsid (r-ZIKV-c), non-structural proteins NS1 (r-ZIKV-NS1), and NS3 (r-ZIKV-NS3) for detection of the infection by ZIKV in blood sera of patients. A commercial polyclonal antibody recognized the r-ZIKV-NS1. Here, among three proteins, NS1 showed the best result for diagnostic purposes using serum samples, despite the high similarity with NS1 from DENV, and could differentiate the infections. The recombinant NS1 was used to produce a monoclonal antibody to differentiate between DENV and ZIKV NS1. As for recombinant proteins, the result for r-ZIKV-NS1 values showed 77% and 100% sensitivity and specificity, respectively, in the IgM assay. Our data showed the protein could successfully differentiate between sera of ZIKV infected patients from sera of those not infected with the virus and differentiate from sera of DENV infected patients. Thus, the generated recombinant proteins have great potential for serological diagnosis of ZIKV in Brazil, where it is indispensable.

摘要

寨卡病毒(ZIKV)感染仅在2015年巴西疫情爆发后才与神经系统疾病相关联。由于缺乏疫苗和精确的诊断方法,因此需要一种精确的方法来检测ZIKV感染。本研究旨在评估三种ZIKV重组蛋白用于ZIKV感染的诊断。在此,我们纯化了稳定的重组ZIKV衣壳蛋白(r-ZIKV-c)、非结构蛋白NS1(r-ZIKV-NS1)和NS3(r-ZIKV-NS3),用于检测患者血清中的ZIKV感染。一种商业多克隆抗体可识别r-ZIKV-NS1。在此,在这三种蛋白中,尽管NS1与登革病毒(DENV)的NS1高度相似,但在使用血清样本进行诊断时,NS1显示出最佳结果,并且能够区分感染情况。重组NS1被用于制备单克隆抗体,以区分DENV和ZIKV的NS1。至于重组蛋白,在IgM检测中,r-ZIKV-NS1值的结果分别显示出77%的灵敏度和100%的特异性。我们的数据表明,该蛋白能够成功区分ZIKV感染患者的血清与未感染该病毒患者的血清,并且能够与DENV感染患者的血清区分开来。因此,所产生的重组蛋白在巴西对ZIKV进行血清学诊断方面具有巨大潜力,在巴西这种诊断是必不可少的。

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