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酵母表达的病毒样颗粒作为抗原检测人血清中人类博卡病毒 1-4 型特异性抗体。

Yeast-generated virus-like particles as antigens for detection of human bocavirus 1-4 specific antibodies in human serum.

机构信息

Institute of Biotechnology, Vilnius University, V. A. Graičiūno 8, 02241, Vilnius, Lithuania.

Clinic of Children's Diseases, Faculty of Medicine, Vilnius University, Vilnius, Lithuania.

出版信息

Appl Microbiol Biotechnol. 2016 Jun;100(11):4935-46. doi: 10.1007/s00253-016-7336-8. Epub 2016 Feb 4.

Abstract

Human bocaviruses (HBoV) are non-enveloped, single-stranded DNA viruses, classified into the genus Bocavirus in the family Parvoviridae. Self-assembled virus-like particles (VLPs) composed of the major capsid protein VP2 of HBoV1-4 and mosaic VLPs composed of both VP2 and VP1 capsid proteins of HBoV1 were generated in yeast Saccharomyces cerevisiae and used to detect HBoV-specific IgG in human serum. Recombinant HBoV VLPs were similar to native HBoV particles in size and morphology. The prevalence of HBoV infection in a group of Lithuanian patients with clinical symptoms of respiratory tract infection was studied using purified yeast-generated VLPs as antigens in a competitive enzyme immunoassay (EIA). After depletion of cross-reactive antibodies, the seroprevalence of HBoV1 was 44.2 % and the seroprevalence of HBoV2-4 was 35.7 %. Mosaic VLPs consisting of HBoV1 VP1 and VP2 proteins showed a stronger reactivity with HBoV1 IgG-positive human serum specimens, and two equivocal serum specimens were reinterpreted as positive. Thus, mosaic VLPs offer a more sensitive tool for HBoV1 serology than currently available serodiagnostics tests based on VP2 VLPs. In conclusion, yeast S. cerevisiae represents an efficient expression system for generating recombinant HBoV1-4 VLPs of diagnostic relevance.

摘要

人博卡病毒(HBoV)是非包膜的单链 DNA 病毒,分类为细小病毒科细小病毒属。自组装的病毒样颗粒(VLPs)由 HBoV1-4 的主要衣壳蛋白 VP2 组成,镶嵌的 VLPs 由 HBoV1 的 VP2 和 VP1 衣壳蛋白组成,在酵母酿酒酵母中生成,并用于检测人血清中的 HBoV 特异性 IgG。重组 HBoV VLP 与天然 HBoV 颗粒在大小和形态上相似。使用纯化的酵母生成的 VLPs 作为抗原,在竞争酶免疫分析(EIA)中研究了一组具有呼吸道感染临床症状的立陶宛患者中 HBoV 感染的流行情况。在耗尽交叉反应性抗体后,HBoV1 的血清流行率为 44.2%,HBoV2-4 的血清流行率为 35.7%。由 HBoV1 VP1 和 VP2 蛋白组成的镶嵌 VLP 与 HBoV1 IgG 阳性的人血清标本显示出更强的反应性,并且两个不确定的血清标本被重新解释为阳性。因此,与目前基于 VP2 VLP 的血清学诊断测试相比,镶嵌 VLP 为 HBoV1 血清学提供了更敏感的工具。总之,酵母酿酒酵母是生成具有诊断相关性的重组 HBoV1-4 VLP 的有效表达系统。

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