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Clinical and Vaccine Immunology, November 2007, p. 1505-1514, Vol. 14, No. 11
1071-412X/07/$08.00+0     doi:10.1128/CVI.00145-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Single Antigen Detects both Immunoglobulin M (IgM) and IgG Antibodies Elicited by All Four Dengue Virus Serotypes ^,

Menaka D. Hapugoda,¹ Gaurav Batra,¹ W. Abeyewickreme,² S. Swaminathan,¹ and N. Khanna^1*

Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, India,¹ Department of Parasitology, Faculty of Medicine, University of Kelaniya, Ragama, Sri Lanka²

Received 4 April 2007/ Returned for modification 25 June 2007/ Accepted 14 September 2007

The resurgence of dengue (DEN) virus infections in the last few decades coupled with the lack of a preventive vaccine and specific antiviral drugs has jointly contributed to making this a significant global public health problem. Currently, symptomatic supportive treatment and fluid replacement therapy are the only means available to minimize DEN-induced mortality. As the clinical symptoms associated with DEN virus infections are indistinguishable from those of many other viral, bacterial, and parasitic infections, specific diagnostic tests assume critical importance in the unequivocal identification of DEN virus infections. We have designed a novel chimeric antigen based on envelope domain III (EDIII), a critical antigenic region of the major structural protein of DEN viruses. We fused EDIIIs corresponding to each of the four DEN virus serotypes using pentaglycyl linkers, overexpressed the resultant tetravalent chimeric protein in Escherichia coli, and affinity purified it in high yields, obtaining 30 mg protein of >95% purity per liter of culture. We show that this tetravalent antigen could specifically recognize anti-DEN virus antibodies of both the immunoglobulin M (IgM) and IgG classes. Using a large panel of IgM antibody capture-enzyme-linked immunosorbent assay- and hemagglutination inhibition-confirmed DEN virus-infected and uninfected patient sera (n = 289), we demonstrate that this tetravalent antigen can function as a diagnostic tool of high sensitivity and specificity.

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* Corresponding author. Mailing address: Recombinant Gene Products Group, International Centre for Genetic Engineering and Biotechnology, P.O. Box 10504, Aruna Asaf Ali Marg, New Delhi 110067, India. Phone: 91-11-26742357, ext. 272. Fax: 91-11-26742316. E-mail: navin{at}icgeb.res.in

Published ahead of print on 26 September 2007.

Supplemental material for this article may be found at http://cvi.asm.org/.

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Clinical and Vaccine Immunology, November 2007, p. 1505-1514, Vol. 14, No. 11
1071-412X/07/$08.00+0     doi:10.1128/CVI.00145-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

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