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Journal of Clinical Microbiology, February 2002, p. 372-375, Vol. 40, No. 2
0095-1137/01/$04.00+0     DOI: 10.1128/JCM.40.2.372-375.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Immunofluorescence Technique Using HeLa Cells Expressing Recombinant Nucleoprotein for Detection of Immunoglobulin G Antibodies to Crimean-Congo Hemorrhagic Fever Virus

Masayuki Saijo,1 Tang Qing,2 Masahiro Niikura,1 Akihiko Maeda,1 Tetsuro Ikegami,1 Koji Sakai,3 Christophe Prehaud,4 Ichiro Kurane,1 and Shigeru Morikawa1*

Special Pathogens Laboratory, Department of Virology 1,1 AIDS Research Center, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan,3 Second Division of Viral Hemorrhagic Fever, Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Changping, Beijing 102206, People's Republic of China,2 Unité de Neuroimmunologie Virale, Institut Pasteur, 75724 Paris Cedex 15, France4

Received 6 August 2001/ Returned for modification 27 September 2001/ Accepted 24 November 2001

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.

* Corresponding author. Mailing address: Special Pathogens Laboratory, Department of Virology 1, National Institute of Infectious Diseases, Gakuen 4-7-1, Musashimurayama, Tokyo 208-0011, Japan. Phone: 81-42-561-0771, ext. 791. Fax: 81-42-564-4881. E-mail: morikawa{at}nih.go.jp.

Journal of Clinical Microbiology, February 2002, p. 372-375, Vol. 40, No. 2
0095-1137/01/$04.00+0     DOI: 10.1128/JCM.40.2.372-375.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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