title: In Vitro Replication Assay for Merkel Cell Polyomavirus (MCPyV).
abstract: Merkel cell polyomavirus (MCPyV) genomes are clonally integrated in tumor cells of ∼95% of all Merkel cell carcinoma (MCC) cases. The virus is highly prevalent; however, where the virus persists and which cell types are permissive for MCPyV replication is still unknown. As a consequence, very little information is available about the life cycle and no fully permissive in vitro replication system has been established. Recently, semi-permissive replication systems based on wild-type MCPyV genomes recovered from the skin of healthy donors or synthetic MCPyV genomes constructed from consensus sequences have been established. The transfection of this intramolecular re-circularized MCPyV DNA into some human cell lines recapitulates efficient DNA replication of the viral genome, viral gene expression as well as moderate levels of virus particle formation. However, serial transmission of infectious virus is still restricted in these cells.
authors.affiliation: Institute for Medical Microbiology, Virology and Hygiene; University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Friederike
authors.initials: F
authors.lastname: Neumann
authors.affiliation: Institute for Medical Microbiology, Virology and Hygiene; University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Manja
authors.initials: M
authors.lastname: Czech-Sioli
authors.affiliation: Heinrich-Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.
authors.forename: Adam
authors.initials: A
authors.lastname: Grundhoff
authors.affiliation: Institute for Medical Microbiology, Virology and Hygiene; University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Fischer
mesh_terms: D002460:Cell Line; D004261:DNA Replication; D006801:Humans; D059965:Merkel cell polyomavirus / Q000235:genetics / Q000502:physiology*; D008967:Molecular Biology / Q000379:methods; D014773:Virology / Q000379:methods; D014779:Virus Replication
keywords: HIRT extract; human polyomavirus; in vitro replication assay; real-time PCR; synthetic MCPyV genome; viral DNA replication