title: Sensitive detection of multiple pathogens using a single DNA probe.
abstract: A simple but promising electrochemical DNA nanosensor was designed, constructed and applied to differentiate a few food-borne pathogens. The DNA probe was initially designed to have a complementary region in Vibrio parahaemolyticus (VP) genome and to make different hybridization patterns with other selected pathogens. The sensor was based on a screen printed carbon electrode (SPCE) modified with polylactide-stabilized gold nanoparticles (PLA-AuNPs) and methylene blue (MB) was employed as the redox indicator binding better to single-stranded DNA. The immobilization and hybridization events were assessed using differential pulse voltammetry (DPV). The fabricated biosensor was able to specifically distinguish complementary, non-complementary and mismatched oligonucleotides. DNA was measured in the range of 2.0×10(-9)-2.0×10(-13)M with a detection limit of 5.3×10(-12)M. The relative standard deviation for 6 replications of DPV measurement of 0.2µM complementary DNA was 4.88%. The fabricated DNA biosensor was considered stable and portable as indicated by a recovery of more than 80% after a storage period of 6 months at 4-45°C. Cross-reactivity studies against various food-borne pathogens showed a reliably sensitive detection of VP.
authors.affiliation: Institute of Advanced Technology, Universiti Putra Malaysia, UPM, 43400 Serdang, Selangor, Malaysia; Food Safety Research Centre, Faculty of Food Science and Technology, Universiti Putra Malaysia, UPM, 43400 Serdang, Selangor, Malaysia.
authors.forename: Noordiana
authors.initials: N
authors.lastname: Nordin
authors.affiliation: Institute of Advanced Technology, Universiti Putra Malaysia, UPM, 43400 Serdang, Selangor, Malaysia; Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, UPM, 43400 Serdang, Selangor, Malaysia. Electronic address: azahy@upm.edu.my.
authors.forename: Nor Azah
authors.initials: NA
authors.lastname: Yusof
authors.affiliation: Institute of Advanced Technology, Universiti Putra Malaysia, UPM, 43400 Serdang, Selangor, Malaysia; Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, UPM, 43400 Serdang, Selangor, Malaysia.
authors.forename: Jaafar
authors.initials: J
authors.lastname: Abdullah
authors.affiliation: Food Safety Research Centre, Faculty of Food Science and Technology, Universiti Putra Malaysia, UPM, 43400 Serdang, Selangor, Malaysia.
authors.forename: Son
authors.initials: S
authors.lastname: Radu
authors.affiliation: La Trobe Institute for Molecular Science, La Trobe University, Victoria 3086, Australia. Electronic address: R.Hushiarian@latrobe.edu.au.
authors.forename: Roozbeh
authors.initials: R
authors.lastname: Hushiarian
mesh_terms: D045424:Complex Mixtures / Q000032:analysis; D003217:Conductometry / Q000295:instrumentation*; D015342:DNA Probes / Q000032:analysis / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005504:Food Analysis / Q000295:instrumentation; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology / Q000295:instrumentation; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles / Q000737:chemistry; D015336:Molecular Probe Techniques / Q000295:instrumentation; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D011091:Polyesters / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014736:Vibrio parahaemolyticus / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D045424:Complex Mixtures; D015342:DNA Probes; D004269:DNA, Bacterial; D011091:Polyesters; C033616:poly(lactide); D006046:Gold
keywords: Electrochemical DNA biosensor; Food-borne pathogens; Methylene blue; Nucleic acid hybridization detection; Polylactide-stabilized gold nanoparticles (PLA-AuNPs)