Publication: Anti-dengue IgG detection by an indirect ELISA.
Issued Date
1995-12-01
Resource Type
ISSN
01251562
Other identifier(s)
2-s2.0-0029422416
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Mahidol University
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SCOPUS
Bibliographic Citation
The Southeast Asian journal of tropical medicine and public health. Vol.26, No.4 (1995), 673-676
Suggested Citation
P. H. Tio, P. Malasit Anti-dengue IgG detection by an indirect ELISA.. The Southeast Asian journal of tropical medicine and public health. Vol.26, No.4 (1995), 673-676. Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/17378
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Title
Anti-dengue IgG detection by an indirect ELISA.
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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%).