Immunofluorescence technique using HeLa cells expressing recombinant nucleoprotein for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus

Masayuki Saijo, Tang Qing, Masahiro Niikura, Akihiko Maeda, Tetsuro Ikegami, Koji Sakai, Christophe Prehaud, Ichiro Kurane, Shigeru Morikawa

Research output: Contribution to journalArticle

67 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)372-375
Number of pages4
JournalJournal of Clinical Microbiology
Volume40
Issue number2
DOIs
StatePublished - 2002
Externally publishedYes

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Crimean-Congo hemorrhagic fever virus
Nucleoproteins
HeLa Cells
Fluorescent Antibody Technique
Immunoglobulin G
Antibodies
Antigens
Vero Cells
Serum
Staining and Labeling
Sensitivity and Specificity
Virus Diseases
Indirect Fluorescent Antibody Technique
Transfection
Epidemiologic Studies

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Immunofluorescence technique using HeLa cells expressing recombinant nucleoprotein for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus. / Saijo, Masayuki; Qing, Tang; Niikura, Masahiro; Maeda, Akihiko; Ikegami, Tetsuro; Sakai, Koji; Prehaud, Christophe; Kurane, Ichiro; Morikawa, Shigeru.

In: Journal of Clinical Microbiology, Vol. 40, No. 2, 2002, p. 372-375.

Research output: Contribution to journalArticle

Saijo, Masayuki ; Qing, Tang ; Niikura, Masahiro ; Maeda, Akihiko ; Ikegami, Tetsuro ; Sakai, Koji ; Prehaud, Christophe ; Kurane, Ichiro ; Morikawa, Shigeru. / Immunofluorescence technique using HeLa cells expressing recombinant nucleoprotein for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus. In: Journal of Clinical Microbiology. 2002 ; Vol. 40, No. 2. pp. 372-375.
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