title: Microfluidics and nanoparticles based amperometric biosensor for the detection of cyanobacteria (Planktothrix agardhii NIVA-CYA 116) DNA.
abstract: Some of the cyanobacteria produce protease inhibitor oligopeptides such as cyanopeptolins and cause drinking water contamination; hence, their detection has great importance to monitor the well-being of water sources that is used for human consumption. In the current study, a fast and sensitive nucleic acid biosensor assay has been described where cyanopeptolin coding region of one of the cyanobacteria (Planktothrix agardhii NIVA-CYA 116) genome has been used as target for monitoring of the fresh water resources. A biochip that has two sets of Au electrode arrays, each consist of shared reference/counter electrodes and 3 working electrodes has been used for the assay. The biochip has been integrated to a microfluidics system and all steps of the assay have been performed during the reagent flow to achieve fast and sensitive DNA detection. On-line hybridization of the target on to the capture probe immobilized surface resulted in a very short assay duration with respect to the conventional static assays. The binding of the avidin and enzyme modified Au nanoparticles to the biotinylated detection probe and the subsequent injection of the substrate enabled a real-time amperometric measurement with a detection limit of 6×10(-12) M target DNA (calibration curve r(2)=0.98). The developed assay enables fast and sensitive detection of cyanopeptolin producing cyanobacteria from freshwater samples and hence shows a promising technology for toxic microorganism detection from environmental samples.
authors.affiliation: Bioelectronic Devices and Systems Group, UEKAE-BILGEM-The Scientific and Technological Research Council of Turkey (TUBITAK), 41470 Gebze, Kocaeli, Turkey; Department of Chemistry, Gebze Technical University, 41400, Gebze, Kocaeli, Turkey.
authors.forename: Zehra
authors.initials: Z
authors.lastname: Ölcer
authors.affiliation: Bioelectronic Devices and Systems Group, UEKAE-BILGEM-The Scientific and Technological Research Council of Turkey (TUBITAK), 41470 Gebze, Kocaeli, Turkey.
authors.forename: Elif
authors.initials: E
authors.lastname: Esen
authors.affiliation: Bioelectronic Devices and Systems Group, UEKAE-BILGEM-The Scientific and Technological Research Council of Turkey (TUBITAK), 41470 Gebze, Kocaeli, Turkey.
authors.forename: Aylin
authors.initials: A
authors.lastname: Ersoy
authors.affiliation: Bioelectronic Devices and Systems Group, UEKAE-BILGEM-The Scientific and Technological Research Council of Turkey (TUBITAK), 41470 Gebze, Kocaeli, Turkey.
authors.forename: Sinan
authors.initials: S
authors.lastname: Budak
authors.affiliation: Fikret Biyal Research Center, Cerrapasa Medical Faculty, Istanbul University, 34098 Istanbul, Turkey.
authors.forename: Dilek
authors.initials: D
authors.lastname: Sever Kaya
authors.affiliation: Bioelectronic Devices and Systems Group, UEKAE-BILGEM-The Scientific and Technological Research Council of Turkey (TUBITAK), 41470 Gebze, Kocaeli, Turkey.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Yağmur Gök
authors.affiliation: Bioelectronic Devices and Systems Group, UEKAE-BILGEM-The Scientific and Technological Research Council of Turkey (TUBITAK), 41470 Gebze, Kocaeli, Turkey.
authors.forename: Serkan
authors.initials: S
authors.lastname: Barut
authors.affiliation: Department of Medical Genetics & REMER, School of Medicine, Medipol University, Beykoz 34810 Istanbul, Turkey.
authors.forename: Duran
authors.initials: D
authors.lastname: Üstek
authors.affiliation: Bioelectronic Devices and Systems Group, UEKAE-BILGEM-The Scientific and Technological Research Council of Turkey (TUBITAK), 41470 Gebze, Kocaeli, Turkey. Electronic address: yildiz.uludag@tubitak.gov.tr.
authors.forename: Yildiz
authors.initials: Y
authors.lastname: Uludag
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D003217:Conductometry / Q000295:instrumentation*; D000458:Cyanobacteria / Q000235:genetics* / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D008839:Microelectrodes; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D006046:Gold
keywords: Amperometry; Cyanobacteria; Cyanopeptolin; Electrochemical sensing; Microfluidics; Real-time electrochemical profiling