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Comparative evaluation of three assays for measurement of dengue virus neutralizing antibodies.
Am J Trop Med Hyg. 2008 Jul; 79(1):115-22.AJ

Abstract

Plaque reduction neutralization tests (PRNTs) are commonly used for measuring levels of dengue virus (DENV) neutralizing antibodies. However, these assays lack a standardized format, generally have a low sample throughput, and are labor-intensive. The objective of the present study was to evaluate two alternative DENV neutralizing antibody assays: an enzyme-linked immunosorbent assay-based microneutralization (MN) assay, and a fluorescent antibody cell sorter-based, DC-SIGN expresser dendritic cell (DC) assay. False-positive rates, serotype specificity, reproducibility, sensitivity, and agreement among the assay methods were assessed using well-characterized but limited numbers of coded test sera. Results showed that all three assays had false-positive rates of less than 10% with titers near the cut-off and generally below the estimated limits of detection. All three methods demonstrated a high degree of specificity and good agreement when used to assay sera and serum mixtures from monovalent vaccinees and sera from patients after primary natural infection, with the only notable exception being moderate-to-high neutralizing antibody titers against DENV 2 measured by PRNT in a mixture containing only DENV 3 and DENV 4 sera. The MN and DC assays demonstrated good reproducibility. All three assays were comparable in their sensitivity, except that the PRNT was less sensitive for measuring DENV 4 antibody, and the MN and DC assays were less sensitive for measuring DENV 2 antibody. However, when used to test sera from persons after tetravalent DENV vaccination or secondary DENV infection, there was poor specificity and poor agreement among the different assays.

Authors+Show Affiliations

Division of Viral Diseases, Walter Reed Army Institute of Research, Silver Spring, Maryland 20910, USA. robert.putnak@na.amedd.army.milNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

18606774

Citation

Putnak, J Robert, et al. "Comparative Evaluation of Three Assays for Measurement of Dengue Virus Neutralizing Antibodies." The American Journal of Tropical Medicine and Hygiene, vol. 79, no. 1, 2008, pp. 115-22.
Putnak JR, de la Barrera R, Burgess T, et al. Comparative evaluation of three assays for measurement of dengue virus neutralizing antibodies. Am J Trop Med Hyg. 2008;79(1):115-22.
Putnak, J. R., de la Barrera, R., Burgess, T., Pardo, J., Dessy, F., Gheysen, D., Lobet, Y., Green, S., Endy, T. P., Thomas, S. J., Eckels, K. H., Innis, B. L., & Sun, W. (2008). Comparative evaluation of three assays for measurement of dengue virus neutralizing antibodies. The American Journal of Tropical Medicine and Hygiene, 79(1), 115-22.
Putnak JR, et al. Comparative Evaluation of Three Assays for Measurement of Dengue Virus Neutralizing Antibodies. Am J Trop Med Hyg. 2008;79(1):115-22. PubMed PMID: 18606774.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparative evaluation of three assays for measurement of dengue virus neutralizing antibodies. AU - Putnak,J Robert, AU - de la Barrera,Rafael, AU - Burgess,Timothy, AU - Pardo,Jorge, AU - Dessy,Francis, AU - Gheysen,Dirk, AU - Lobet,Yves, AU - Green,Sharone, AU - Endy,Timothy P, AU - Thomas,Stephen J, AU - Eckels,Kenneth H, AU - Innis,Bruce L, AU - Sun,Wellington, PY - 2008/7/9/pubmed PY - 2008/8/9/medline PY - 2008/7/9/entrez SP - 115 EP - 22 JF - The American journal of tropical medicine and hygiene JO - Am J Trop Med Hyg VL - 79 IS - 1 N2 - Plaque reduction neutralization tests (PRNTs) are commonly used for measuring levels of dengue virus (DENV) neutralizing antibodies. However, these assays lack a standardized format, generally have a low sample throughput, and are labor-intensive. The objective of the present study was to evaluate two alternative DENV neutralizing antibody assays: an enzyme-linked immunosorbent assay-based microneutralization (MN) assay, and a fluorescent antibody cell sorter-based, DC-SIGN expresser dendritic cell (DC) assay. False-positive rates, serotype specificity, reproducibility, sensitivity, and agreement among the assay methods were assessed using well-characterized but limited numbers of coded test sera. Results showed that all three assays had false-positive rates of less than 10% with titers near the cut-off and generally below the estimated limits of detection. All three methods demonstrated a high degree of specificity and good agreement when used to assay sera and serum mixtures from monovalent vaccinees and sera from patients after primary natural infection, with the only notable exception being moderate-to-high neutralizing antibody titers against DENV 2 measured by PRNT in a mixture containing only DENV 3 and DENV 4 sera. The MN and DC assays demonstrated good reproducibility. All three assays were comparable in their sensitivity, except that the PRNT was less sensitive for measuring DENV 4 antibody, and the MN and DC assays were less sensitive for measuring DENV 2 antibody. However, when used to test sera from persons after tetravalent DENV vaccination or secondary DENV infection, there was poor specificity and poor agreement among the different assays. SN - 1476-1645 UR - https://www.unboundmedicine.com/medline/citation/18606774/Comparative_evaluation_of_three_assays_for_measurement_of_dengue_virus_neutralizing_antibodies_ L2 - https://www.lens.org/lens/search/patent/list?q=citation_id:18606774 DB - PRIME DP - Unbound Medicine ER -