A novel fluorescence-based array biosensor: principle and application to DNA hybridization assays.
Biosensors and Bioelectronics. 2008-02-01; 23(7): 987-994
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1. Biosens Bioelectron. 2008 Feb 28;23(7):987-94. doi: 10.1016/j.bios.2007.10.006.
Epub 2007 Oct 22.
A novel fluorescence-based array biosensor: principle and application to DNA
Schultz E(1), Galland R, Du Bouëtiez D, Flahaut T, Planat-Chrétien A, Lesbre F,
Hoang A, Volland H, Perraut F.
(1)Laboratoire d’Imagerie et des Système d’Acquisition, Commissariat à l’Energie
Atomique, 17 Rue des Martyrs, Grenoble, France.
A novel fluorescence-based array biosensor targeted for field applications, such
as environmental monitoring, has been developed, and successfully applied to DNA
hybridization assays. The purpose was to meet the demand for automated, portable
but easy-to-maintain systems allowing continuous flow monitoring of surface
reactions. The biosensor presented here can be distinguished from the existing
systems by the optical method used, which provides an enhanced simplicity and
robustness, and enables a simple maintenance by potentially unskilled personnel.
The system is based on a conventional microscope slide which acts both as
transducer and biological array sensor. The excited fluorescence is guided by
total internal reflection into the slide to the detector which is directly
interfaced to the slide. Each region of the sensor array is successively
optically interrogated, and the detection of the corresponding fluorescent
emission synchronized. A real-time three-analyte analysis is thus feasible
without any mechanical scanning movement or optical imaging systems as generally
used in the existing instruments. The ability of the biosensor to operate in
continuous flow for several tens of hours has been demonstrated. The biosensor
has been assessed in terms of stability, and slide-to-slide reproducibility,
which is found to be less than 3.7%, thus far below the standard biological
reproducibility. DNA hybridization assays were performed to estimate a limit of
detection, which was found to be 16 mol/microm(2), and to determine the reaction
kinetics associated to the DNA model used. The developed biosensor is thus shown
to be able to predict reaction kinetics, and to monitor in real time surface
reactions between targets and probes.
PMID: 18207730 [Indexed for MEDLINE]