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

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.
PMID
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Authors

Mayor MeshTerms

Biosensing Techniques

Keywords

Malaria

PfHRP 2

biosensor

immunosensor

nanoparticles

parasites

Journal Title biosensors
Publication Year Start




PMID- 28718841
OWN - NLM
STAT- MEDLINE
DA  - 20170718
DCOM- 20170731
LR  - 20170731
IS  - 2079-6374 (Electronic)
IS  - 2079-6374 (Linking)
VI  - 7
IP  - 3
DP  - 2017 Jul 18
TI  - Development of an Immunosensor for PfHRP 2 as a Biomarker for Malaria Detection.
LID - E28 [pii]
LID - 10.3390/bios7030028 [doi]
AB  - 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)/H(2)O(2) 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 ngmL-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 pgmL-1 was achieved in buffer samples and 40 pgmL-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.
FAU - Hemben, Aver
AU  - Hemben A
AD  - Surface Engineering and Nanotechnology Institute, Cranfield University,
      Cranfield, Bedfordshire MK43 0AL, UK. [email protected]
FAU - Ashley, Jon
AU  - Ashley J
AD  - Surface Engineering and Nanotechnology Institute, Cranfield University,
      Cranfield, Bedfordshire MK43 0AL, UK. [email protected]
AD  - Department of Micro- and Nanotechnology, Technical University of Denmark,
      Produktionstorvet, 2800 Kgs. Lyngby, Denmark. [email protected]
FAU - Tothill, Ibtisam E
AU  - Tothill IE
AD  - Surface Engineering and Nanotechnology Institute, Cranfield University,
      Cranfield, Bedfordshire MK43 0AL, UK. [email protected]
LA  - eng
PT  - Journal Article
DEP - 20170718
PL  - Switzerland
TA  - Biosensors (Basel)
JT  - Biosensors
JID - 101609191
RN  - 0 (Antigens, Protozoan)
RN  - 0 (Biomarkers)
RN  - 0 (HRP-2 antigen, Plasmodium falciparum)
RN  - 0 (Proteins)
RN  - 0 (Protozoan Proteins)
RN  - 0 (histidine-rich proteins)
RN  - 7440-57-5 (Gold)
SB  - IM
MH  - Antigens, Protozoan/*blood
MH  - Biomarkers/*blood
MH  - *Biosensing Techniques
MH  - Gold/chemistry
MH  - Humans
MH  - Malaria, Falciparum/*blood/parasitology
MH  - Plasmodium falciparum/isolation & purification/pathogenicity
MH  - Proteins/*isolation & purification
MH  - Protozoan Proteins/*blood
OTO - NOTNLM
OT  - Malaria
OT  - PfHRP 2
OT  - biosensor
OT  - immunosensor
OT  - nanoparticles
OT  - parasites
COI - The authors declare no conflict of interest.
EDAT- 2017/07/19 06:00
MHDA- 2017/08/02 06:00
CRDT- 2017/07/19 06:00
PHST- 2017/06/13 [received]
PHST- 2017/07/10 [revised]
PHST- 2017/07/12 [accepted]
AID - bios7030028 [pii]
AID - 10.3390/bios7030028 [doi]
PST - epublish
SO  - Biosensors (Basel). 2017 Jul 18;7(3). pii: E28. doi: 10.3390/bios7030028.