期刊
BIOSENSORS & BIOELECTRONICS
卷 54, 期 -, 页码 428-434出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2013.11.023
关键词
Human papillomavirus; acpcPNA; Anthraquinone; Electrochemical detection; Screen-printed electrode
类别
资金
- Science Achievement Scholarship of Thailand (SAST)
- Center of Innovative Nanotechnology (CIN)
- Electrochemistry and Optical Spectroscopy Research Unit
- Program in Petrochemical & Polymer Science, and Organic Synthesis Research Unit, Chulalongkorn University
- Thailand Research Fund [BRG5380008, RTA5280002]
- Thai Government Stimulus Package 2 under the Project for Establishment of Comprehensive Center for Innovative Food, Health Products and Agriculture [TKK2555]
- Chulalongkorn University
- National Research University Project of CHE
- Ratch adaphiseksomphot Endowment Fund [AM1009I-56, AM1006A-56]
An electrochemical biosensor based on an immobilized anthraquinone-labeled pyrrolidinyl peptide nucleic acid (acpcPNA) probe was successfully developed for the selective detection of human papillomavirus (HPV) type 16 DNA. A 14-mer acpcPNA capture probe was designed to recognize a specific 14 nucleotide region of HPV type 16 L1 gene. The redox-active label anthraquinone (AQ) was covalently attached to the N-terminus of the acpcPNA probe through an amide bond. The probe was immobilized onto a chitosan-modified disposable screen-printed carbon electrode via a C-terminal lysine residue using glutaraldehyde as a crosslinking agent. Hybridization with the target DNA was studied by measuring the electrochemical signal response of the AQ label using square-wave voltammetric analysis. The calibration curve exhibited a linear range between 0.02 and 12.0 mu M with a limit of detection and limit of quantitation of 4 and 14 nM, respectively. This DNA sensing platform was successfully applied to detect the HPV type 16 DNA from a PCR amplified (240 bp fragment of the L1 gene) sample derived from the HPV type 16 positive human cancer cell line (SiHa), and failed to detect the HPV-negative c33a cell line. The sensor probe exhibited very high selectivity for the complementary 14 base oligonucleotide over the non-complementary oligonucleotides with sequences derived from HPV types 18, 31 and 33. The proposed sensor provides an inexpensive tool for the early stage detection of HPV type 16, which is an important biomarker for cervical cancer. (C) 2013 Elsevier B.V. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据