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Anti-dengue IgG detection by an indirect ELISA.

Authors: P H, Tio; P, Malasit;

Anti-dengue IgG detection by an indirect ELISA.

Abstract

Protein-free culture media were originally developed for hybridomas to simplify downstream processing and purification. For the same reasons, we have used these protein-free media for passaging dengue 2 virus in C6/36 cells. This provided us with an infected supernatant (DenPF) which could then be used as coating antigens for an indirect enzyme-linked immunosorbent assay (ELISA) to determine dengue IgG levels. Using this preparation, the main immunogenic band as seen by immunoblot appeared to be viral envelope protein (E). Without the high concentrations of "competing" proteins from fetal calf serum (FCS), the Den2PF could be directly coated onto 96-well ELISA plates. The amount of viral proteins in Den2PF appeared to be sufficient so that there was no need for further purification steps, eg polyethylene glycol (PEG) precipitation, which made this preparation cost effective. It compared favorably with the dengue dot enzyme immunoassay (DEIA; sensitivity of 95.7% and specificity of 95.2%).

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Keywords

Cost-Benefit Analysis, Reproducibility of Results, Enzyme-Linked Immunosorbent Assay, Dengue Virus, Antibodies, Viral, Sensitivity and Specificity, Culture Media, Serum-Free, Dengue, Immunoenzyme Techniques, Case-Control Studies, Immunoglobulin G, Humans

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
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