Nik Nurhanan Nik Mansor, Tan Toh Leong, Eka Safitri, Dedi Futra, Nurul Saadah Ahmad, Dian Nasriana Nasuruddin, Azlin Itnin, Ida Zarina Zaini, Khaizurin Tajul Arifin, Lee Yook Heng, Nurul Izzaty Hassan
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. © 2018 by the authors. Licensee MDPI, Basel, Switzerland.
School of Chemical Sciences and Food Technology, Faculty of Science & Technology, Universiti Kebangsaan Malaysia, Bangi, 43600, Selangor DE, Malaysia; Department of Emergency Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras, 56000, Kuala Lumpur, Malaysia; Department of Chemistry, Faculty of Mathematics and Natural Sciences, Syiah Kuala University (USK), Darussalam-Banda Acheh, 23111, Indonesia; The Department of Chemistry Education, Faculty of Education, Universitas Riau, Kampus Binawidya KM 12.5, Pekanbaru, 28293, Riau, Indonesia; Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras, 56000, Kuala Lumpur, Malaysia; Department of Biochemistry Education, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras, 56000, Kuala Lumpur, Malaysia
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