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

Abstract

BackgroundThe reproducibilty of dengue IgM and IgG ELISA was studied in serum and filter paper blood spots from Vietnamese febrile patients.Methods781 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.ResultsCases 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.ConclusionDengue 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.

Trends in the patterns of IgM and IgG antibodies in febrile persons with suspected dengue in Barbados, an English-speaking Caribbean country, 2006–2013

“…The pattern of seroconversion seen among a select population of children attending the medical facility for gastrointestinal symptoms in the Dominican Republic [12] and among febrile persons who were screened for dengue in Jamaica [14] was similar to the pattern seen in this study. Interestingly, the pattern of IgG seroprevalence seen in our study was also similar to the patterns of high IgG seroprevalence that were reported in population-based prospective studies from some of the other highly endemic countries in the Americas [11,[22][23][24][25] and those of other highly endemic countries in southeast Asia [26][27][28].…”

Evaluation of immunological markers in serum, filter-paper blood spots, and saliva for dengue diagnosis and epidemiological studies

“…This is important because filter-paper blood spots are easier to obtain than venous blood in infants and young children. Although filter-paper specimens have been used for detecting anti-DENV IgM, 22,23 no reports describe measurement of anti-DENV IgA in this sample type. Even though the sensitivity of anti-DENV IgM detection in saliva was lower than that of IgM and IgA in serum and filter-paper blood spots, it could potentially be useful for epidemiological surveillance and in confirmation of dengue in an outbreak situation in a particular community.…”

Maternal SerumChlamydia PneumoniaeAntibodies and CRP Levels in Women with Preeclampsia and Gestational Hypertension

“…There was no difference in the storage length between the study groups. However, even the long storage period does not have any remarkable effect on the IgG antibody (53) or hsCRP levels (54).…”