Electrochemical genosensor for specific detection of the food-borne pathogen, Vibrio cholerae.
A disposable horseradish peroxidase (HRP)-based electrochemical genosensor was developed for chronoamperometric detection of single-stranded asymmetric lolB gene PCR amplicon (118 bp in length) of the food-borne pathogen, Vibrio cholerae. A two-step sandwich-type hybridization strategy using two specific probes was employed for specific detection of the target single-stranded DNA (ssDNA). The analytical performances of the detection platform have been evaluated using a synthetic ssDNA (ST3) which was identical to the target single-stranded amplicon and a total of 19 bacterial strains. Under optimal condition, ST3 was calibrated with a dynamic range of 0.4883-15.6250 nM. By coupling asymmetric PCR amplification, the probe-based electrochemical genosensor was highly specific to the target organism (100% specificity) and able to detect as little as 0.85 ng/μl of V. cholerae genomic DNA.
Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, 16150, Kubang Kerian, Kelantan, Malaysia.
Kim-Fatt
KF
Low
Kritsanaporn
K
Chuenrangsikul
Patsamon
P
Rijiravanich
Werasak
W
Surareungchai
Yean-Yean
YY
Chan
D015374:Biosensing Techniques / Q000379:methods*; D055664:Electrochemical Techniques / Q000379:methods*; D005516:Food Microbiology; D006735:Horseradish Peroxidase / Q000378:metabolism; D009693:Nucleic Acid Hybridization / Q000379:methods; D012680:Sensitivity and Specificity; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification*
D006735:Horseradish Peroxidase; C021372:horseradish apoperoxidase