Rapid Identification of Dengue Virus Isolates by using Monoclonal Antibodies in an Indirect Immunofluorescence Assay

Erik A. HenchalDepartments of Virus Diseases and Biochemistry, Walter Reed Army Institute of Research, Washington, D.C. 20012

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J. M. McCownDepartments of Virus Diseases and Biochemistry, Walter Reed Army Institute of Research, Washington, D.C. 20012

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M. C. SeguinDepartments of Virus Diseases and Biochemistry, Walter Reed Army Institute of Research, Washington, D.C. 20012

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M. K. GentryDepartments of Virus Diseases and Biochemistry, Walter Reed Army Institute of Research, Washington, D.C. 20012

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W. E. BrandtDepartments of Virus Diseases and Biochemistry, Walter Reed Army Institute of Research, Washington, D.C. 20012

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Type-specific monoclonal antibodies prepared against the four dengue (DEN) virus serotypes were evaluated for their ability to identify low-passage human and mosquito isolates from Jamaica and West Africa by an indirect immunofluorescence assay. Serotyped human isolates from Jamaican dengue fever patients included 12 DEN-1, two DEN-2, and five DEN-4 viruses. Viruses from West Africa included 84 DEN-2 mosquito strains as well as two DEN-1 and one DEN-2 from humans. Results obtained using the immunofluorescence assay were consistent with virus identifications obtained using the more classical but costly and time-consuming plaque-reduction neutralization test. More viral isolates and higher virus yields were obtained using the C6/36 clone of Aedes albopictus cells rather than LLC-MK2 (monkey kidney) cells. Dengue type-specific monoclonal antibodies detected prototype viral antigens 24–48 hours postinfection in C6/36 cells. This is the first time that monoclonal antibodies have been used to serotype low-passage flavivirus isolates.

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