Development of an Immunosensor for PfHRP 2 as a Biomarker for Malaria Detection
| Autor: | Jon Ashley, Aver Hemben, Ibtisam E. Tothill |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
lcsh:Biotechnology
Plasmodium falciparum 030231 tropical medicine Clinical Biochemistry Protozoan Proteins Antigens Protozoan Biosensing Techniques Immunosensor parasites biosensor 01 natural sciences Article 03 medical and health sciences 0302 clinical medicine SDG 3 - Good Health and Well-being lcsh:TP248.13-248.65 medicine Humans Parasites Malaria Falciparum immunosensor Detection limit biology Chemistry 010401 analytical chemistry Proteins PfHRP 2 General Medicine Elisa assay Serum samples biology.organism_classification medicine.disease Molecular biology Malaria 0104 chemical sciences nanoparticles Colloidal gold Nanoparticles Biomarker (medicine) Gold Biosensor Biomarkers |
| Zdroj: | Biosensors, Vol 7, Iss 3, p 28 (2017) Biosensors Biosensors; Volume 7; Issue 3; Pages: 28 Hemben, A, Ashley, J & Tothill, I E 2017, ' Development of an Immunosensor for P f HRP 2 as a Biomarker for Malaria Detection ', Biosensors, vol. 7, no. 3 . https://doi.org/10.3390/bios7030028 |
| ISSN: | 2079-6374 |
| DOI: | 10.3390/bios7030028 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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