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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Gold nanoparticle-based fluorescence immunoassay for malaria antigen detection
Analytical and Bioanalytical Chemistry, Volume 402, No. 3, Year 2012
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Description
The development of rapid detection assays for malaria diagnostics is an area of intensive research, as the traditional microscopic analysis of blood smears is cumbersome and requires skilled personnel. Here, we describe a simple and sensitive immunoassay that successfully detects malaria antigens in infected blood cultures. This homogeneous assay is based on the fluorescence quenching of cyanine 3B (Cy3B)-labeled recombinant Plasmodium falciparum heat shock protein 70 (PfHsp70) upon binding to gold nanoparticles (AuNPs) functionalized with an anti-Hsp70 monoclonal antibody. Upon competition with the free antigen, the Cy3B-labeled recombinant PfHsp70 is released to solution resulting in an increase of fluorescence intensity. Two types of AuNP-antibody conjugates were used as probes, one obtained by electrostatic adsorption of the antibody on AuNPs surface and the other by covalent bonding using protein cross-linking agents. In comparison with cross-linked antibodies, electrostatic adsorption of the antibodies to the AuNPs surfaces generated conjugates with increased activity and linearity of response, within a range of antigen concentration from 8.2 to 23.8 μg.mL -1. The estimated LOD for the assay is 2.4 μg.mL -1 and the LOQ is 7.3 μg.mL -1. The fluorescence immunoassay was successfully applied to the detection of antigen in malaria-infected human blood cultures at a 3% parasitemia level, and is assumed to detect parasite densities as low as 1,000 parasites.μL -1. © 2011 Springer-Verlag.
Authors & Co-Authors
Guirgis, Bassem Shenoda
Portugal, Caparica
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa
Egypt, New Cairo
School of Sciences and Engineering
Sá E Cunha, Cláudia
Portugal, Lisbon
Faculdade de Medicina, Universidade de Lisboa
Gomes, Inês
Portugal, Caparica
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa
Portugal, Lisbon
Faculdade de Medicina, Universidade de Lisboa
Cavadas, Miguel
Portugal, Caparica
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa
Silva, Isabel
Portugal, Caparica
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa
Dória, Gonçalo
Portugal, Caparica
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa
Blatch, Gregory Lloyd
South Africa, Grahamstown
Rhodes University
Australia, Melbourne
Victoria University
Baptista, Pedro V.
Portugal, Caparica
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa
Pereira, Eulália
Portugal, Porto
Universidade do Porto
Azzazy, Hassan Mohamed El Said
Egypt, New Cairo
School of Sciences and Engineering
Mota, Maria M.
Portugal, Lisbon
Faculdade de Medicina, Universidade de Lisboa
Prudêncio, Miguel
Portugal, Lisbon
Faculdade de Medicina, Universidade de Lisboa
Franco, Ricardo
Portugal, Caparica
Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa
Statistics
Citations: 79
Authors: 13
Affiliations: 6
Identifiers
Doi:
10.1007/s00216-011-5489-y
ISSN:
16182642
Research Areas
Infectious Diseases