PMID- 29912194
OWN - NLM
STAT- MEDLINE
DCOM- 20180730
LR  - 20200603
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 136
DP  - 2018 Jun 3
TI  - Development of an Electrochemical DNA Biosensor to Detect a Foodborne Pathogen.
LID - 10.3791/56585 [doi]
LID - 56585
AB  - Vibrio parahaemolyticus (V. parahaemolyticus) is a common foodborne pathogen that 
      contributes to a large proportion of public health problems globally, 
      significantly affecting the rate of human mortality and morbidity. Conventional 
      methods for the detection of V. parahaemolyticus such as culture-based methods, 
      immunological assays, and molecular-based methods require complicated sample 
      handling and are time-consuming, tedious, and costly. Recently, biosensors have 
      proven to be a promising and comprehensive detection method with the advantages 
      of fast detection, cost-effectiveness, and practicality. This research focuses on 
      developing a rapid method of detecting V. parahaemolyticus with high selectivity 
      and sensitivity using the principles of DNA hybridization. In the work, 
      characterization of synthesized polylactic acid-stabilized gold nanoparticles 
      (PLA-AuNPs) was achieved using X-ray Diffraction (XRD), Ultraviolet-visible 
      Spectroscopy (UV-Vis), Transmission Electron Microscopy (TEM), Field-emission 
      Scanning Electron Microscopy (FESEM), and Cyclic Voltammetry (CV). We also 
      carried out further testing of stability, sensitivity, and reproducibility of the 
      PLA-AuNPs. We found that the PLA-AuNPs formed a sound structure of stabilized 
      nanoparticles in aqueous solution. We also observed that the sensitivity improved 
      as a result of the smaller charge transfer resistance (Rct) value and an increase 
      of active surface area (0.41 cm(2)). The development of our DNA biosensor was 
      based on modification of a screen-printed carbon electrode (SPCE) with PLA-AuNPs 
      and using methylene blue (MB) as the redox indicator. We assessed the 
      immobilization and hybridization events by differential pulse voltammetry (DPV). 
      We found that complementary, non-complementary, and mismatched oligonucleotides 
      were specifically distinguished by the fabricated biosensor. It also showed 
      reliably sensitive detection in cross-reactivity studies against various 
      food-borne pathogens and in the identification of V. parahaemolyticus in fresh 
      cockles.
FAU - Nordin, Noordiana
AU  - Nordin N
AD  - Laboratory of Food Safety and Food Integrity, Institute of Tropical Agriculture 
      and Food Security, Universiti Putra Malaysia; noordiana@upm.edu.my.
FAU - Yusof, Nor Azah
AU  - Yusof NA
AD  - Laboratory of Functional Device, Institute of Advanced Technology, Universiti 
      Putra Malaysia; Department of Chemistry, Faculty of Science, Universiti Putra 
      Malaysia.
FAU - Radu, Son
AU  - Radu S
AD  - Laboratory of Food Safety and Food Integrity, Institute of Tropical Agriculture 
      and Food Security, Universiti Putra Malaysia.
FAU - Hushiarian, Roozbeh
AU  - Hushiarian R
AD  - La Trobe Institute for Molecular Science, La Trobe University; 
      r.hushiarian@latrobe.edu.au.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20180603
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Biosensing Techniques/*methods
MH  - DNA/*genetics
MH  - Electrochemistry/*methods
MH  - Electrodes/*statistics & numerical data
MH  - Foodborne Diseases/*diagnosis/pathology
MH  - Humans
MH  - Vibrio parahaemolyticus/*pathogenicity
PMC - PMC6101519
EDAT- 2018/06/19 06:00
MHDA- 2018/07/31 06:00
PMCR- 2020/06/03
CRDT- 2018/06/19 06:00
PHST- 2018/06/19 06:00 [entrez]
PHST- 2018/06/19 06:00 [pubmed]
PHST- 2018/07/31 06:00 [medline]
PHST- 2020/06/03 00:00 [pmc-release]
AID - 56585 [pii]
AID - 10.3791/56585 [doi]
PST - epublish
SO  - J Vis Exp. 2018 Jun 3;(136):56585. doi: 10.3791/56585.