title: Immunofluorescence technique using HeLa cells expressing recombinant nucleoprotein for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus.
abstract: A HeLa cell line continuously expressing recombinant nucleoprotein (rNP) of the Crimean-Congo hemorrhagic fever virus (CCHFV) was established by transfection with an expression vector containing the cDNA of CCHFV NP (pKS336-CCHFV-NP). These cells were used as antigens for indirect immunofluorescence (IF) to detect immunoglobulin G antibodies to CCHFV. The sensitivity and specificity of this IF technique were examined by using serum samples and were compared to those of the IF technique using CCHFV-infected Vero E6 cells (authentic antigen). Staining of the CCHFV rNP expressed in HeLa cells showed a unique granular pattern similar to that of CCHFV-infected Vero E6 cells. Positive staining could easily be distinguished from a negative result. All 13 serum samples determined to be positive by using the authentic antigen were also determined to be positive by using CCHFV rNP-expressing HeLa cells (recombinant antigen). The 108 serum samples determined to be negative by using the authentic antigen were also determined to be negative by using the recombinant antigen. Thus, both the sensitivity and the specificity of this IF technique were 100% compared to the IF with authentic antigen. The novel IF technique using CCHFV rNP-expressing HeLa cells can be used not only for diagnosis of CCHF but also for epidemiological studies on CCHFV infections.
authors.affiliation: Special Pathogens Laboratory, Department of Virology 1. AIDS Research Center, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Saijo
authors.forename: Tang
authors.initials: T
authors.lastname: Qing
authors.forename: Masahiro
authors.initials: M
authors.lastname: Niikura
authors.forename: Akihiko
authors.initials: A
authors.lastname: Maeda
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Ikegami
authors.forename: Koji
authors.initials: K
authors.lastname: Sakai
authors.forename: Christophe
authors.initials: C
authors.lastname: Prehaud
authors.forename: Ichiro
authors.initials: I
authors.lastname: Kurane
authors.forename: Shigeru
authors.initials: S
authors.lastname: Morikawa
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D002522:Chlorocebus aethiops; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D003225:Hemorrhagic Fever Virus, Crimean-Congo / Q000276:immunology*; D006479:Hemorrhagic Fever, Crimean / Q000175:diagnosis* / Q000453:epidemiology; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D009698:Nucleoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011995:Recombination, Genetic; D012680:Sensitivity and Specificity; D014709:Vero Cells
chemical_list: D000914:Antibodies, Viral; D007074:Immunoglobulin G; D009698:Nucleoproteins