title: Specific detection of Mycobacterium sp. genomic DNA using dual labeled gold nanoparticle based electrochemical biosensor.
abstract: The present study was aimed at the development and evaluation of a DNA electrochemical biosensor for Mycobacterium sp. genomic DNA detection in a clinical specimen using a signal amplifier as dual-labeled AuNPs. The DNA electrochemical biosensors were fabricated using a sandwich detection strategy involving two kinds of DNA probes specific to Mycobacterium sp. genomic DNA. The probes of enzyme ALP and the detector probe both conjugated on the AuNPs and subsequently hybridized with target DNA immobilized in a SAM/ITO electrode followed by characterization with CV, EIS, and DPV analysis using the electroactive species para-nitrophenol generated by ALP through hydrolysis of para-nitrophenol phosphate. The effect of enhanced sensitivity was obtained due to the AuNPs carrying numerous ALPs per hybridization and a detection limit of 1.25 ng/ml genomic DNA was determined under optimized conditions. The dual-labeled AuNP-facilitated electrochemical sensor was also evaluated by clinical sputum samples, showing a higher sensitivity and specificity and the outcome was in agreement with the PCR analysis. In conclusion, the developed electrochemical sensor demonstrated unique sensitivity and specificity for both genomic DNA and sputum samples and can be employed as a regular diagnostics tool for Mycobacterium sp. monitoring in clinical samples.
authors.affiliation: Nanocomposites Research Group, Department of Chemical Engineering, Anna University, Chennai, TN 600 025, India.
authors.forename: Chinnasamy
authors.initials: C
authors.lastname: Thiruppathiraja
authors.forename: Senthilkumar
authors.initials: S
authors.lastname: Kamatchiammal
authors.forename: Periyakaruppan
authors.initials: P
authors.lastname: Adaikkappan
authors.forename: Devakirubakaran Jayakar
authors.initials: DJ
authors.lastname: Santhosh
authors.forename: Muthukaruppan
authors.initials: M
authors.lastname: Alagar
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D001681:Biological Assay; D015374:Biosensing Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics*; D058266:Dielectric Spectroscopy; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D016680:Genome, Bacterial / Q000235:genetics*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D015335:Molecular Probes / Q000737:chemistry; D009161:Mycobacterium / Q000235:genetics*; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D013194:Staining and Labeling; D017971:Tin Compounds / Q000737:chemistry
chemical_list: D004269:DNA, Bacterial; D015335:Molecular Probes; D017971:Tin Compounds; C109984:indium tin oxide; D006046:Gold; D000469:Alkaline Phosphatase