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Sensors 2018, 18(3), 686; https://doi.org/10.3390/s18030686

An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme System

1
School of Chemical Sciences and Food Technology, Faculty of Science & Technology, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor DE, Malaysia
2
Department of Emergency Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia
3
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Syiah Kuala University (USK), Darussalam-Banda Acheh 23111, Indonesia
4
The Department of Chemistry Education, Faculty of Education, Universitas Riau, Kampus Binawidya KM 12.5, Pekanbaru 28293, Riau, Indonesia
5
Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia
6
Department of Biochemistry Education, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, Malaysia
*
Author to whom correspondence should be addressed.
Received: 28 December 2017 / Revised: 1 February 2018 / Accepted: 17 February 2018 / Published: 26 February 2018
(This article belongs to the Special Issue Development of Enzymatic Electrochemical Biosensors and Applications)
Full-Text   |   PDF [3742 KB, uploaded 26 February 2018]   |  

Abstract

A tri-enzyme system consisting of choline kinase/choline oxidase/horseradish peroxidase was used in the rapid and specific determination of the biomarker for bacterial sepsis infection, secretory phospholipase Group 2-IIA (sPLA2-IIA). These enzymes were individually immobilized onto the acrylic microspheres via succinimide groups for the preparation of an electrochemical biosensor. The reaction of sPLA2-IIA with its substrate initiated a cascading enzymatic reaction in the tri-enzyme system that led to the final production of hydrogen peroxide, which presence was indicated by the redox characteristics of potassium ferricyanide, K3Fe(CN)6. An amperometric biosensor based on enzyme conjugated acrylic microspheres and gold nanoparticles composite coated onto a carbon-paste screen printed electrode (SPE) was fabricated and the current measurement was performed at a low potential of 0.20 V. This enzymatic biosensor gave a linear range 0.01–100 ng/mL (R2 = 0.98304) with a detection limit recorded at 5 × 10−3 ng/mL towards sPLA2-IIA. Moreover, the biosensor showed good reproducibility (relative standard deviation (RSD) of 3.04% (n = 5). The biosensor response was reliable up to 25 days of storage at 4 °C. Analysis of human serum samples for sPLA2-IIA indicated that the biosensor has potential for rapid bacterial sepsis diagnosis in hospital emergency department. View Full-Text
Keywords: amperometric biosensor; choline kinase; choline oxidase; horseradish peroxidase; sepsis; bacterial infection; sPLA2-IIA amperometric biosensor; choline kinase; choline oxidase; horseradish peroxidase; sepsis; bacterial infection; sPLA2-IIA
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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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Nik Mansor, N.N.; Leong, T.T.; Safitri, E.; Futra, D.; Ahmad, N.S.; Nasuruddin, D.N.; Itnin, A.; Zaini, I.Z.; Arifin, K.T.; Heng, L.Y.; Hassan, N.I. An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme System. Sensors 2018, 18, 686.

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