Portable Electrochemical DNA Sensors Based on Gene Amplification Reactions to Screen and Identify Pathogen and SNPs.

Abstract

In this paper, we introduce portable sensors based on genetic measurements that can be used in the field for the diagnosis of infectious diseases and disease risk based on SNPs (single nucleotide polymorphisms). In particular, the sensors are based on electrochemical measurements that can be performed with printed electrodes and small measuring devices. Indicator molecules that can bind to nucleic acid molecules in various ways are already known, and some of these molecules have electrochemical activity. First, we investigated the change in their electrochemical responses in a solution system. As a result, we searched for nucleic acid-binding molecules whose current value changes in the presence of DNA. In addition, when we measured the change in the current value, associated with the amplification of specific genes, such as PCR (polymerase chain reaction) and LAMP (loop-mediated isothermal amplification), we found that the current value decreased with the number of amplifications, indicating that specific genes can be monitored electrochemically. Based on this principle, we showed that pathogenic microorganisms and viruses, such as Salmonella, O157 E. coli, hepatitis B virus, periodontal disease bacteria, antibiotic-resistant bacteria and influenza virus, were able to be measured. The method was also applied to the diagnosis of SNPs, such as ApoE (apolipoprotein E), which is a risk factor for Alzheimer’s disease. Rapid PCR was available with a microfluidic device, and a simple method was also presented with the isothermal amplification of LAMP.