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ABSTRACT

A rapid (<7-min) immunochromatographic test for immunoglobulin M (IgM) and IgG antibodies to dengue viruses was evaluated by using hospital admission and discharge sera from 124 patients. The reference laboratory diagnosis was based on the results of virus isolation, hemagglutination-inhibition assay (HAI), and enzyme immunoassay (EIA). By the standard assays, patients experienced primary dengue virus infection (n = 30), secondary dengue virus infection (n = 48), Japanese encephalitis (JE) virus infection (n = 20), or no flavivirus infection (n = 26). The rapid test demonstrated 100% sensitivity in the diagnosis of dengue virus infection and was able to distinguish between primary and secondary dengue virus infections through the separate determinations of IgM and IgG. For all patients with primary dengue virus infection a positive test for IgM to dengue virus and a negative test for IgG to dengue virus were obtained, whereas for 46 of 48 patients (96%) with secondary dengue virus infection, a positive test for IgG to dengue virus with or without a positive test for IgM to dengue virus was obtained. The remaining two patients with secondary dengue virus infection had positive IgM test results and negative IgG test results. Furthermore, the rapid test was positive for patients confirmed to be infected with different dengue virus serotypes (12 infected with dengue virus serotype 1, 4 infected with dengue virus serotype 2, 3 infected with dengue virus serotype 3, and 2 infected with dengue virus serotype 4). The specificity of the test for nonflavivirus infections was 88% (3 of 26 positive), while for JE virus infections the specificity of the test was only 50% (10 of 20). However, most patients with secondary dengue virus infection were positive for both IgM and IgG antibodies to dengue virus, while no patients with JE virus infection had this profile, so cross-reactivity was only a concern for a small proportion of patients with secondary dengue infections. The rapid test demonstrated a good correlation with the reference EIA and HAI and should be useful for the rapid diagnosis of dengue virus infections.

MATERIALS AND METHODS

 

Case definitions.

In children experiencing a febrile illness consistent with dengue fever or DHF, dengue virus infections were defined as the isolation of a dengue virus, the detection of IgM to dengue virus (as opposed to IgM to Japanese encephalitis [JE] virus), or a sustained elevation (≥1:2,560) or a fourfold rise in dengue virus hemagglutination-inhibition titer. JE virus infection was defined as a febrile illness associated with a decrease in consciousness and the presence of IgM to JE virus in the cerebrospinal fluid. Dengue virus infection was categorized as primary or secondary according to the World Health Organization criteria (29) and the standard operating procedure for the reference EIA (10).

rapid immunochromatographic gmat test peanut
    rapid immunochromatographic gmat test peanut

Serum samples.

Serum samples were collected from patients at the time of hospital admission and the time of discharge at either the Queen Sirikit National Institute of Child Health (Bangkok Children’s Hospital); the Kamphaeng Phet Provincial Hospital, Thailand; or the Centre for Tropical Diseases, Ho Chi Minh City, Vietnam. The serum was frozen at −70°C prior to assay. In this study we used paired serum specimens from 124 patients, representing patients with primary dengue virus infection (n = 30), secondary dengue virus infection (n = 48), JE virus infection (n = 20), or no evidence of flavivirus infection (n = 26).

HAI Acetone-extracted sera were tested for antibodies by HAI as described previously (6), except that the assay was modified to a microtiter plate format. Dengue virus types 1 to 4 and JE virus (8 U each) were used. Antigens were produced by sucrose acetone extraction of the brains of suckling mice infected with the following prototype mouse-adapted virus strains: DEN-1 Hawaii, DEN-2 New Guinea C, DEN-3 H-87, DEN-4 H-241, and JE virus Nakayama. A fourfold increase was considered positive for acute flavivirus infection. The infection was diagnosed as a primary infection if the titers a week or more after the onset of illness were less than or equal to 1:1,280 or as a secondary infection if antibody titers were greater than 1:1,280.

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