Electrochemical detection of a Vibrio parahaemolyticus sequence-specific gene based on a gold electrode modified with a single stranded probe

Abstract

An electrochemical DNA biosensor for specific-sequences detection of Vibrio parahaemolyticus (VP) was fabricated. A singlestranded 20-mer oligonucleotide (ssDNA) and 6-mercapto-1-hexanol (MCH) were immobilized via a thiol linker on gold disk electrodes by self-assembling. The ssDNA underwent hybridization in a hybridization solution containing complementary or non-complementary or single base pair mismatched DNA sequences of VP. Examination of changes in response to these three target DNAs showed that the developed biosensor had a high selectivity and sensitivity.

Single-walled carbon nanotubes–carboxyl-functionalized graphene oxide-based electrochemical DNA biosensor for thermolabile hemolysin gene detection

“…Vibrio parahaemolyticus, a gram-negative bacterium distributed throughout the estuarine environment, is considered to be the source of acute gastroenteritis and some cases of septicemia in humans. [1][2][3] The infection of Vibrio parahaemolyticus usually occurs when consuming raw or undercooked seafood. 4 The thermolabile hemolysin (tlh) gene is considered to be a useful target for the detection of Vibrio parahaemolyticus because it was conrmed to exit in all of the Vibrio parahaemolyticus stains identied to date.…”

Electrochemical Identification of DNA Single Nucleotide Polymorphisms (SNPs) Based on Different Redox Probes

“…Electrochemical sensors have the advantages for SNPs analysis, such as high sensitivity, low cost, rapid response, and easy miniaturization, which are widely applied to detect DNA SNPs by the researchers. [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17] For constructing electrochemical sensors (Scheme 1), usually thiol-modified probe DNA (p-DNA) is assembled on Au surface to form p-DNA self-assembled monolayers (SAMs), and hybridized with target DNA (complementary DNA, abbreviated by c-DNA or SNPs mutant DNA, abbreviated by m-DNA) in solution. The hybridization of p-DNA SAMs on Au with c-DNA or m-DNA in solution will arouse different changes of DNA surface charge, spatial configuration and packing, which can be detected using some redox probes such as Ru(NH 3 ) 6 3+ , Fe(CN) 6 3−/4− , methylene blue (MB) or daunorubicin (DM) by electrochemical methods (cyclic voltammetry, differential pulse voltammetry, electrochemical impedance spectroscopy etc).…”

“…However, Wilson et al 7 reported that AC mismatch might be identified well using Ru(NH 3 ) 6 3+ ; 2) most of literatures about detecting DNA SNPs have not reported the surface coverage of p-DNA (g p-DNA ) on Au. 5,[7][8][9][10][11][12][13][14][15] The g p-DNA of p-DNA on Au is a key factor in DNA SNPs identification because the permeability and diffusion of redox probes through DNA SAMs before or after hybridization are closely related to g p-DNA ; 3) what is the relationship of redox probes' types with DNA SNPs identification effect? Interaction of Ru(NH 3 ) 6 3+ or Fe(CN) 6 3−/4− with DNA is mainly electrostatic force.…”

“…a Self-assembly of double-stranded DNA (ds-DNA) directly on Au; 3,4,9,13 Firstly self-assembly of p-DNA on Au, and then hybridization with target DNA. [5][6][7][8][10][11][12][14][15][16] Some abbreviations: Au nanoparticles (AuNPs), poly(3,4-ethylenedioxythiophene) (PEDOT), glass carbon electrode (GCE), graphene (GR), glutaraldehyde (GTD), chitosan-mutiwalled carbon nanotubes (CS-MWNTs), graphene -chitosan (GR-CS).…”