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Journal of Clinical Microbiology, August 2006, p. 2872-2878, Vol. 44, No. 8
0095-1137/06/$08.00+0     doi:10.1128/JCM.00777-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Serotype 1-Specific Monoclonal Antibody-Based Antigen Capture Immunoassay for Detection of Circulating Nonstructural Protein NS1: Implications for Early Diagnosis and Serotyping of Dengue Virus Infections

Hua Xu,^1, Biao Di,^2, Yu-xian Pan,¹ Li-wen Qiu,¹ Ya-di Wang,¹ Wei Hao,¹ Li-juan He,² Kwok-yung Yuen,³ and Xiao-yan Che^1*

Center of Laboratory, Zhujiang Hospital, Southern Medical University,¹ Center for Disease Control and Prevention of Guangzhou, Guangzhou,² Department of Microbiology, The University of Hong Kong, Hong Kong Special Administrative Region, People's Republic of China³

Received 12 April 2006/ Returned for modification 16 May 2006/ Accepted 24 May 2006

Rapid diagnosis and serotyping of dengue virus (DV) infections are important for timely clinical management and epidemiological control in areas where multiple flaviviruses are endemic. However, the speed and accuracy of diagnosis must be balanced against test cost and availability, especially in developing countries. We developed a specific antigen capture enzyme-linked immunosorbent assay (ELISA) for early detection and serotyping of DV serotype 1 (DV1) by using well-characterized monoclonal antibodies (MAbs) specific to nonstructural protein 1 (NS1) of DV1. With this assay, a total of 462 serum specimens from clinically probable DV1-infected patients during the DV1 epidemic in Guangdong, China, in 2002 and 2003 were analyzed. DV1 NS1 was detectable in blood circulation from the first day up to day 18 after onset of symptoms, with a peak at days 6 to 10. The sensitivity of DV1 NS1 detection in serum specimens with reference to results from reverse transcriptase PCR was 82%, and the specificity was 98.9% with reference to 469 healthy blood donors. No cross-reactions with any of the other three DV serotypes or other closely related members of the genus Flavivirus (Japanese encephalitis virus and Yellow fever virus) were observed when tested with the clinical specimens or virus cultures. These findings suggest that the serotype-specific MAb-based NS1 antigen capture ELISA may be a valuable tool for early diagnosis and serotyping of DV infections, while also providing a standardized assay for the analysis of a great number of clinical samples with convenience and cost-effectiveness.

Hua Xu and Biao Di contributed equally to this work.

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