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Journal of Clinical Microbiology, March 2001, p. 977-982, Vol. 39, No. 3
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.3.977-982.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification of B-Cell Epitope of Dengue Virus Type 1 and Its Application in Diagnosis of Patients

Han-Chung Wu,1,* Yue-Ling Huang,1 Ting-Ting Chao,1 Jia-Tsrong Jan,1 Jau-Ling Huang,1 Hsien-Yuan Chiang,1 Chwan-Chuen King,2 and Men-Fang Shaio1

Institute of Preventive Medicine, National Defense Medical Center,1 and Institute of Epidemiology, National Taiwan University,2 Taipei, Taiwan, Republic of China

Received 8 June 2000/Returned for modification 21 September 2000/Accepted 19 December 2000

Using a serotype-specific monoclonal antibody (MAb) of dengue virus type 1 (DEN-1), 15F3-1, we identified the B-cell epitope of DEN-1 from a random peptide library displayed on phage. Fourteen immunopositive phage clones that bound specifically to MAb 15F3-1 were selected. These phage-borne peptides had a consensus motif of HxYaWb (a = S/T, b = K/H/R) that mimicked the sequence HKYSWK, which corresponded to amino acid residues 111 to 116 of the nonstructural protein 1 (NS1) of DEN-1. Among the four synthetic peptides corresponding to amino acid residues 110 to 117 of the NS1 of DEN-1, -2, -3, and -4, only one peptide, EHKYSWKS (P14M) of DEN-1, was found to bind to 15F3-1 specifically. Furthermore, P14M was shown to inhibit the binding of phage particles to 15F3-1 in a competitive inhibition assay. Histidine111 (His111) was crucial to the binding of P14M to 15F3-1, since its binding activity dramatically reduced when it changed to leucine111 (Leu111). This epitope-based peptide demonstrated its clinical diagnostic potential when it reacted with a high degree of specificity with serum samples obtained from both DEN-1-infected rabbits and patients. Based on these observations, our DEN-1 epitope-based serologic test could be useful in laboratory viral diagnosis and in understanding the pathogenesis of DEN-1.

* Corresponding author. Mailing address: Institute of Preventive Medicine, National Defense Medical Center, P.O. Box 90048-700, San-Hsia, Taiwan. Phone: 886-2-2673-2230. Fax: 886-2-2673-6994. E-mail: ipmc3{at}ms29.hinet.net.

Journal of Clinical Microbiology, March 2001, p. 977-982, Vol. 39, No. 3
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.3.977-982.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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