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Quantitative and sensitive detection of immune-complexed and free HIV antigen after boiling of serum.


Schüpbach, J; Böni, J (1993). Quantitative and sensitive detection of immune-complexed and free HIV antigen after boiling of serum. Journal of Virological Methods, 43(2):247-256.

Abstract

5-min boiling of 3-fold diluted serum leads to the quantitative release of immune-complexed HIV antigen (Ag) that can be readily measured. The method is capable of detecting as little as 4 to 5 pg/ml Ag p24 depending on the Ag test used. Of a panel of 50 anti-HIV-positive sera, 30 (60%) were Ag-positive after heat denaturation, 17 (34%; P < or = 0.01) after acidification, and 13 (26%; P < or = 0.001) when tested undenatured. The superiority of heat denaturation was greatest in sera with high-titered antibodies to p24. Occurrence and mean concentrations of heat-denatured Ag were reversely correlated with p24-specific antibody. It is suggested that heat-denatured proteins be used for raising antibodies that will serve as capture or trace antibodies in Ag assays. This simple change in the manufacturing of test reagents in combination with a short boil before testing may solve the problem of incomplete Ag detection due to immune complex formation.

Abstract

5-min boiling of 3-fold diluted serum leads to the quantitative release of immune-complexed HIV antigen (Ag) that can be readily measured. The method is capable of detecting as little as 4 to 5 pg/ml Ag p24 depending on the Ag test used. Of a panel of 50 anti-HIV-positive sera, 30 (60%) were Ag-positive after heat denaturation, 17 (34%; P < or = 0.01) after acidification, and 13 (26%; P < or = 0.001) when tested undenatured. The superiority of heat denaturation was greatest in sera with high-titered antibodies to p24. Occurrence and mean concentrations of heat-denatured Ag were reversely correlated with p24-specific antibody. It is suggested that heat-denatured proteins be used for raising antibodies that will serve as capture or trace antibodies in Ag assays. This simple change in the manufacturing of test reagents in combination with a short boil before testing may solve the problem of incomplete Ag detection due to immune complex formation.

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Additional indexing

Item Type:Journal Article, refereed
Communities & Collections:04 Faculty of Medicine > Institute of Medical Virology
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:English
Date:1 July 1993
Deposited On:11 Feb 2008 12:28
Last Modified:05 Apr 2016 12:22
Publisher:Elsevier
ISSN:0166-0934
Publisher DOI:https://doi.org/10.1016/0166-0934(93)90080-B
PubMed ID:8366170

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