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Biosensors and Bioelectronics
Vol. 77, 2016, Pages: 805–811

Non-protein coding RNA-based genosensor with quantum dots as electrochemical labels for attomolar detection of multiple pathogens

Dinesh Vijian, Suresh V. Chinni, Lee Su Yin, Benchaporn Lertanantawong, Werasak Surareungchai

Department of Biotechnology, Faculty of Applied Sciences, AIMST University, 08100 Kedah, Malaysia.

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 choleraeSalmonella sp. andShigella 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.

Keywords: Electrochemical biosensor; Enteric pathogens; Quantum dots; Salmonella sp.; Shigella sp.; Vibrio cholerae.

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