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Biosensors 2017, 7(3), 28; doi:10.3390/bios7030028

Development of an Immunosensor for PfHRP 2 as a Biomarker for Malaria Detection

1
Surface Engineering and Nanotechnology Institute, Cranfield University, Cranfield, Bedfordshire MK43 0AL, UK
2
Department of Micro- and Nanotechnology, Technical University of Denmark, Produktionstorvet, 2800 Kgs. Lyngby, Denmark
*
Author to whom correspondence should be addressed.
Received: 13 June 2017 / Revised: 10 July 2017 / Accepted: 12 July 2017 / Published: 18 July 2017
(This article belongs to the Special Issue Biosensors for the Detection of Biomarkers)
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Abstract

Plasmodium falciparum histidine-rich protein 2 (PfHRP 2) was selected in this work as the biomarker for the detection and diagnosis of malaria. An enzyme-linked immunosorbent assay (ELISA) was first developed to evaluate the immunoreagent’s suitability for the sensor’s development. A gold-based sensor with an integrated counter and an Ag/AgCl reference electrode was first selected and characterised and then used to develop the immunosensor for PfHRP 2, which enables a low cost, easy to use, and sensitive biosensor for malaria diagnosis. The sensor was applied to immobilise the anti-PfHRP 2 monoclonal antibody as the capture receptor. A sandwich ELISA assay format was constructed using horseradish peroxidase (HRP) as the enzyme label, and the electrochemical signal was generated using a 3, 3′, 5, 5′tetramethyl-benzidine dihydrochloride (TMB)/H2O2 system. The performance of the assay and the sensor were optimised and characterised, achieving a PfHRP 2 limit of detection (LOD) of 2.14 ng·mL−1 in buffer samples and 2.95 ng∙mL−1 in 100% spiked serum samples. The assay signal was then amplified using gold nanoparticles conjugated detection antibody-enzyme and a detection limit of 36 pg∙mL−1 was achieved in buffer samples and 40 pg∙mL−1 in serum samples. This sensor format is ideal for malaria detection and on-site analysis as a point-of-care device (POC) in resource-limited settings where the implementation of malaria diagnostics is essential in control and elimination efforts. View Full-Text
Keywords: Malaria; PfHRP 2; parasites; immunosensor; biosensor; nanoparticles Malaria; PfHRP 2; parasites; immunosensor; biosensor; nanoparticles
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Hemben, A.; Ashley, J.; Tothill, I.E. Development of an Immunosensor for PfHRP 2 as a Biomarker for Malaria Detection. Biosensors 2017, 7, 28.

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