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Linares, E.M. ; Pannuti, C.S.* ; Kubota, L.T.* ; Thalhammer, S.

Immunospot assay based on fluorescent nanoparticles for dengue fever detection.

Biosens. Bioelectron. 41, 180-185 (2013)
DOI PMC
Open Access Green as soon as Postprint is submitted to ZB.
Dengue fever is one of the most neglected tropical diseases and of highest international public health importance, with 50 million cases worldwide every year. Early detection can decrease mortality rates from more than 20% to less than 1% and the relevant early diagnosis analyte is the viral non-structural glycoprotein, NS1. Currently, enzyme linked immunosorbent assay (ELISA) is the method of choice to detect NS1. However, this is a time consuming method, requiring 3-5h, and it is the bottleneck for routine of clinical analysis laboratory in epidemic periods, when hundreds of samples should be tested. Here we describe an easy method combining principles of fluorophore linked immunosorbent assay (FLISA) and enzyme linked immunospotting (ELISPOT). For detection, we used mouse anti-NS1 IgG labeled with fluorescent nanoparticles. The presented procedure needs only 4μL of serum samples and requires 45-60min. The detection limit, 5.2ng/mL, is comparable to ELISA tests. The comparison of 83 samples with a commercial ELISA revealed a sensitivity of 81% and specificity of 88%. The use of fluorescent nanoparticles provides a higher sensitivity than an assay using usual fluorescent dye molecules, besides avoiding bleaching effects. Based on the results, the proposed method provides fast, specific and sensitive results, and proves to be a suitable method for Dengue NS1 detection in impoverished regions or epidemic areas.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Immunospot Assay ; Dengue Fever ; Fluorescent Nanoparticles ; Elisa; Linked-immunosorbent-assay ; Dot Enzyme-immunoassay ; Protein Ns1 ; Hemorrhagic-fever ; Virus ; Diagnosis ; Antigen ; Antibodies ; Infections ; Illness
ISSN (print) / ISBN 0956-5663
e-ISSN 1873-4235
Quellenangaben Volume: 41, Issue: 1, Pages: 180-185 Article Number: , Supplement: ,
Publisher Elsevier
Non-patent literature Publications
Reviewing status Peer reviewed