Electrophoresis-Enhanced Detection of Deoxyribonucleic Acids on a Membrane-Based Lateral Flow Strip Using Avian Influenza H5 Genetic Sequence as the Model

Sensors

Jui-Chuang Wu¹  Chih-Hung Chen²  Ja-Wei Fu² 

[1] Department of Chemical Engineering, Chung Yuan Christian University, Chung Li, Tao Yuan 32023, Taiwan;

关键词: membrane-based lateral-flow detection;    DNA detection;    electrophoresis;    avian influenza;    biosensors;   

DOI  :  10.3390/s140304399

来源: mdpi

【 摘 要 】

This study reports a simple strategy to detect a deoxyribonucleic acid (DNA) on a membrane-based lateral flow (MBLF) strip without tedious gel preparation, gel electrophoresis, and EtBr-staining processes. The method also enhances the detection signal of the genetic sample. A direct electric field was applied over two ends of the MBLF strips to induce an electrophoresis of DNAs through the strips. The signal enhancement was demonstrated by the detection of the H5 subtype of avian influenza virus (H5 AIV). This approach showed an excellent selectivity of H5 AIV from other two control species, Arabidopsis thaliana and human PSMA5. It also showed an effective signal repeatability and sensitivity over a series of analyte concentrations. Its detection limit could be enhanced, from 40 ng to 0.1 ng by applying 12 V. The nano-gold particles for the color development were labeled on the capture antibody, and UV-VIS and TEM were used to check if the labeling was successful. This detection strategy could be further developed to apply on the detection of drug-allergic genes at clinics or detection of infectious substances at incident sites by a simple manipulation with an aid of a mini-PCR machine and auxiliary kits.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.