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. 2006 Jan 25:6:13.
doi: 10.1186/1471-2334-6-13.

Enzyme-linked immunoassay for dengue virus IgM and IgG antibodies in serum and filter paper blood

Thanh Nga T Tran  1 , Peter J de Vries, Lan Phuong Hoang, Giao T Phan, Hung Q Le, Binh Q Tran, Chi Mai T Vo, Nam V Nguyen, Piet A Kager, Nico Nagelkerke, Jan Groen
Affiliations

Enzyme-linked immunoassay for dengue virus IgM and IgG antibodies in serum and filter paper blood

Thanh Nga T Tran et al. BMC Infect Dis. .

Abstract

Background: The reproducibilty of dengue IgM and IgG ELISA was studied in serum and filter paper blood spots from Vietnamese febrile patients.

Methods: 781 pairs of acute (t0) and convalescent sera, obtained after three weeks (t3) and 161 corresponding pairs of filter paper blood spots were tested with ELISA for dengue IgG and IgM. 74 serum pairs were tested again in another laboratory with similar methods, after a mean of 252 days.

Results: Cases were classified as no dengue (10 %), past dengue (55%) acute primary (7%) or secondary (28%) dengue. Significant differences between the two laboratories' results were found leading to different diagnostic classification (kappa 0.46, p < 0.001). Filter paper results correlated poorly to serum values, being more variable and lower with a mean (95% CI) difference of 0.82 (0.36 to 1.28) for IgMt3, 0.94 (0.51 to 1.37) for IgGt0 and 0.26 (-0.20 to 0.71) for IgGt3. This also led to differences in diagnostic classification (kappa value 0.44, p < 0.001) The duration of storage of frozen serum and dried filter papers, sealed in nylon bags in an air-conditioned room, had no significant effect on the ELISA results.

Conclusion: Dengue virus IgG antibodies in serum and filter papers was not affected by duration of storage, but was subject to inter-laboratory variability. Dengue virus IgM antibodies measured in serum reconstituted from blood spots on filter papers were lower than in serum, in particular in the acute phase of disease. Therefore this method limits its value for diagnostic confirmation of individual patients with dengue virus infections. However the detection of dengue virus IgG antibodies eluted from filter paper can be used for sero-prevalence cross sectional studies.

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Figures

Figure 1
Figure 1
Comparison of dengue ELISA in serum and reconstituted filter paper blood. Serum and blood spots on filter papers were collected simultaneously from the same patient and tested at the same time with the same ELISA. The reference lines indicate the cut off value for the index value (< 1: negative; ≥ 1: positive). The sloped lines indicate the regression of y to x (_____) and x to y (-----).
See this image and copyright information in PMC

References

    1. Guzman MG, Kouri G. Dengue diagnosis, advances and challenges. Int J Infect Dis. 2004;8:69–80. doi: 10.1016/j.ijid.2003.03.003. - DOI - PubMed
    1. Vaughn DW, Nisalak A, Solomon T, Kalayanarooj S, Nguyen MD, Kneen R, Cuzzubbo A, Devine PL. Rapid serologic diagnosis of dengue virus infection using a commercial capture ELISA that distinguishes primary and secondary infections. Am J Trop Med Hyg. 1999;60:693–698. - PubMed
    1. Groen J, Koraka P, Velzing J, Copra C, Osterhaus AD. Evaluation of six immunoassays for detection of dengue virus-specific immunoglobulin M and G antibodies. Clin Diagn Lab Immunol. 2000;7:867–871. doi: 10.1128/CDLI.7.6.867-871.2000. - DOI - PMC - PubMed
    1. Koraka P, Zeller H, Niedrig M, Osterhaus AD, Groen J. Reactivity of serum samples from patients with a flavivirus infection measured by immunofluorescence assay and ELISA. Microbes Infect. 2002;4:1209–1215. doi: 10.1016/S1286-4579(02)01647-7. - DOI - PubMed
    1. Innis BL, Nisalak A, Nimmannitya S, Kusalerdchariya S, Chongswasdi V, Suntayakorn S, Puttisri P, Hoke CH. An enzyme-linked immunosorbent assay to characterize dengue infections where dengue and Japanese encephalitis co-circulate. Am J Trop Med Hyg. 1989;40:418–427. - PubMed

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