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

• Published in: Applied microbiology and biotechnology 2016-06, Vol.100 (11), p.4935-4946
• Main Authors: Tamošiūnas, Paulius Lukas, Petraitytė-Burneikienė, Rasa, Bulavaitė, Aistė, Marcinkevičiūtė, Kornelija, Simutis, Karolis, Lasickienė, Rita, Firantienė, Regina, Ėmužytė, Regina, Žvirblienė, Aurelija, Sasnauskas, Kęstutis
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Aged; Analysis; Animals; Antibodies; Antibodies, Viral - blood; Antigen-antibody reactions; Antigens; Antigens, Viral - immunology; Biomedical and Life Sciences; Biotechnologically Relevant Enzymes and Proteins; Biotechnology; Capsid Proteins - immunology; Child; Child, Preschool; Cloning; Cloning, Molecular; Cross Reactions; Electrophoresis, Polyacrylamide Gel; Enzymes; Epidemiology; Escherichia coli - genetics; Escherichia coli - metabolism; Female; Health aspects; Host-virus relationships; Human bocavirus - isolation & purification; Humans; Immunoglobulin G - blood; Infant; Life Sciences; Lithuania; Male; Mice; Mice, Inbred BALB C; Microbial Genetics and Genomics; Microbiology; Middle Aged; Mutagenesis; Observations; Parvoviridae; Parvoviridae Infections - blood; Parvoviridae Infections - diagnosis; Parvoviridae Infections - immunology; Parvoviruses; Plasmids; Proteins; Recombinant Proteins - immunology; Recombinant Proteins - metabolism; Respiratory tract; Saccharomyces cerevisiae; Saccharomyces cerevisiae - metabolism; Serology; Specimen Handling; Studies; Viruses; Yeast; Yeasts; Young Adult
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-016-7336-8

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.