An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme System
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 micr...
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| Format: | Article |
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MDPI AG
2018-02-01
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| Series: | Sensors |
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| Online Access: | http://www.mdpi.com/1424-8220/18/3/686 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Nik Nurhanan Nik Mansor |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T21:46:13Z |
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| id | doaj.art-d82ca240e58f4e6eb6f1e7033bf70eef |
| institution | Directory Open Access Journal |
| issn | 1424-8220 |
| language | English |
| last_indexed | 2024-04-11T21:46:13Z |
| publishDate | 2018-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Sensors |
| spelling | doaj.art-d82ca240e58f4e6eb6f1e7033bf70eef2022-12-22T04:01:25ZengMDPI AGSensors1424-82202018-02-0118368610.3390/s18030686s18030686An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme SystemNik Nurhanan Nik Mansor0Tan Toh Leong1Eka Safitri2Dedi Futra3Nurul Saadah Ahmad4Dian Nasriana Nasuruddin5Azlin Itnin6Ida Zarina Zaini7Khaizurin Tajul Arifin8Lee Yook Heng9Nurul Izzaty Hassan10School of Chemical Sciences and Food Technology, Faculty of Science & Technology, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor DE, MalaysiaDepartment of Emergency Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, MalaysiaDepartment of Chemistry, Faculty of Mathematics and Natural Sciences, Syiah Kuala University (USK), Darussalam-Banda Acheh 23111, IndonesiaThe Department of Chemistry Education, Faculty of Education, Universitas Riau, Kampus Binawidya KM 12.5, Pekanbaru 28293, Riau, IndonesiaDepartment of Emergency Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, MalaysiaDepartment of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, MalaysiaDepartment of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, MalaysiaDepartment of Emergency Medicine, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, MalaysiaDepartment of Biochemistry Education, Faculty of Medicine, Universiti Kebangsaan Malaysia, Cheras 56000, Kuala Lumpur, MalaysiaSchool of Chemical Sciences and Food Technology, Faculty of Science & Technology, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor DE, MalaysiaSchool of Chemical Sciences and Food Technology, Faculty of Science & Technology, Universiti Kebangsaan Malaysia, Bangi 43600, Selangor DE, MalaysiaA 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.http://www.mdpi.com/1424-8220/18/3/686amperometric biosensorcholine kinasecholine oxidasehorseradish peroxidasesepsisbacterial infectionsPLA2-IIA |
| spellingShingle | 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 An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme System Sensors amperometric biosensor choline kinase choline oxidase horseradish peroxidase sepsis bacterial infection sPLA2-IIA |
| title | An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme System |
| title_full | An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme System |
| title_fullStr | An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme System |
| title_full_unstemmed | An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme System |
| title_short | An Amperometric Biosensor for the Determination of Bacterial Sepsis Biomarker, Secretory Phospholipase Group 2-IIA Using a Tri-Enzyme System |
| title_sort | amperometric biosensor for the determination of bacterial sepsis biomarker secretory phospholipase group 2 iia using a tri enzyme system |
| topic | amperometric biosensor choline kinase choline oxidase horseradish peroxidase sepsis bacterial infection sPLA2-IIA |
| url | http://www.mdpi.com/1424-8220/18/3/686 |
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