Institute of Biotechnology, Vilnius University, V. A. Graičiūno 8, 02241, Vilnius, Lithuania.
Clinic of Children's Diseases, Faculty of Medicine, Vilnius University, Vilnius, Lithuania.
Appl Microbiol Biotechnol. 2016 Jun;100(11):4935-46. doi: 10.1007/s00253-016-7336-8. Epub 2016 Feb 4.
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.
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。