title: Non-protein coding RNA-based genosensor with quantum dots as electrochemical labels for attomolar detection of multiple pathogens.
abstract: The ability of a diagnostic test to detect multiple pathogens simultaneously is useful to obtain meaningful information for clinical treatment and preventive measures. We report a highly sensitive and specific electrochemical biosensor assay for simultaneous detection of three gene targets using quantum dots (QDs). The targets are novel non-protein coding RNA (npcRNA) sequences of Vibrio cholerae, Salmonella sp. and Shigella sp., which cause diarrheal diseases. QDs (PbS, CdS, ZnS) were synthesized and functionalized with DNA probes that were specific to each pathogen. Electrochemical detection of QDs was performed using square wave anodic stripping voltammetry (SWASV). The QDs gave distinct peaks at 0.5 V (PbS), 0.75 V (CdS) and 1.1 V (ZnS). There was no interference in signal response when all three QDs were mixed and detected simultaneously. The detection limits of single and multiplex assays with linear targets and PCR products were in the attomolar ranges. The high assay sensitivity, in combination with specific npcRNA sequences as novel diagnostic targets, makes it a viable tool for detecting pathogens from food, environment and clinical samples.
authors.affiliation: Department of Biotechnology, Faculty of Applied Sciences, AIMST University, 08100 Kedah, Malaysia.
authors.forename: Dinesh
authors.initials: D
authors.lastname: Vijian
authors.affiliation: Department of Biotechnology, Faculty of Applied Sciences, AIMST University, 08100 Kedah, Malaysia.
authors.forename: Suresh V
authors.initials: SV
authors.lastname: Chinni
authors.affiliation: Department of Biotechnology, Faculty of Applied Sciences, AIMST University, 08100 Kedah, Malaysia.
authors.forename: Lee Su
authors.initials: LS
authors.lastname: Yin
authors.affiliation: Pilot Plant Development and Training Institute, King Mongkut's University of Technology Thonburi, Bangkok, Thailand. Electronic address: l_benjap@yahaoo.com.
authors.forename: Benchaporn
authors.initials: B
authors.lastname: Lertanantawong
authors.affiliation: School of Bioresources and Technology, King Mongkut's University of Technology Thonburi, Bangkok, Thailand.
authors.forename: Werasak
authors.initials: W
authors.lastname: Surareungchai
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation; D003217:Conductometry / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D008832:Microchemistry / Q000295:instrumentation*; D045663:Quantum Dots; D012329:RNA, Bacterial / Q000032:analysis* / Q000235:genetics*; D022661:RNA, Untranslated / Q000737:chemistry* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling
chemical_list: D012329:RNA, Bacterial; D022661:RNA, Untranslated
keywords: Electrochemical biosensor; Enteric pathogens; Quantum dots; Salmonella sp.; Shigella sp.; Vibrio cholerae