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Title
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Proof of principle: an HIV p24 microsphere immunoassay with potential application to HIV clinical diagnosis
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Author
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Abstract
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| The measurement of CD4 counts and viral loads on a single instrument such as an affordable flow cytometer could considerably reduce the cost related to the follow-up of antiretroviral therapy in resource-poor settings. The aim of this study was to assess whether the HIV-1 p24 antigen could be measured using a microsphere-based flow cytometric (FC) assay and the experimental conditions necessary for processing plasma samples. A commercial anti-p24 antibody pair from Biomaric was used to develop a p24 microsphere immunoassay (MIA) using HIV culture supernatant as the source of antigen. The ultrasensitive Perkin Elmer enzyme immunoassay (EIA) served as a reference assay. Quantification of HIV p24 using the heat-mediated immune complex disruption format described for plasma samples was feasible using the Biomaric MIA and applicable to a broad range of HIV-1 Group M subtypes. The inclusion of a tyramide amplification step was successful and increased the fluorescence signal up to 3 logs as compared with the MIA without amplification. The analytical sensitivity of this ultrasensitive Biomaric assay reached 1 pg/mL, whereas the ultrasensitive Perkin Elmer EIA was sensitive to less than 0.17 pg/mL. Our data indicate, for the first time, that the principle of p24 detection using the heat-denatured ultrasensitive format can be applied to FC. |
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Language
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English
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Source (journal)
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Cytometry: part B: clinical cytometry. - New York, 2003, currens
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Publication
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New York
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2009
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ISSN
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1552-4949
[print]
1552-4957
[online]
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DOI
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10.1002/CYTO.B.20466
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Volume/pages
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76
:3
(2009)
, p. 231-236
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ISI
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000265371500011
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Full text (Publisher's DOI)
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