Colorimetric silver detection of DNA microarrays.
Development of microarrays has revolutionized gene expression analysis and molecular diagnosis through miniaturization and the multiparametric features. Critical factors affecting detection efficiency of targets hybridization on microarray are the design of capture probes, the way they are attached to the support, and the sensitivity of the detection method. Microarrays are currently detected in fluorescence using a sophisticated confocal laser-based scanner. In this work, we present a new colorimetric detection method which is intented to make the use of microarray a powerful procedure and a low-cost tool in research and clinical settings. The signal generated with this method results from the precipitation of silver onto nanogold particles bound to streptavidin, the latter being used for detecting biotinylated DNA. This colorimetric method has been compared to the Cy-3 fluorescence method. The detection limit of both methods was equivalent and corresponds to 1 amol of biotinylated DNA attached on an array. Scanning and data analysis of the array were obtained with a colorimetric-based workstation.
Alexandre
I
I
Laboratoire de Biochimie Cellulaire, Facultés Universitaires Notre Dame de la Paix, 61 Rue de Bruxelles, Namur, 5000, Belgium. isabelle.alexandre@fundp.ac.be
Hamels
S
S
Dufour
S
S
Collet
J
J
Zammatteo
N
N
De Longueville
F
F
Gala
J L
JL
Remacle
J
J
D001426:Bacterial Proteins / Q000235:genetics; D019921:Biotinylation; D011232:Chemical Precipitation; D003124:Colorimetry / Q000191:economics / Q000379:methods*; D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000378:metabolism; D006046:Gold / Q000032:analysis / Q000737:chemistry; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics / Q000379:methods*; D010084:Oxidation-Reduction; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012834:Silver / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D013210:Staphylococcus / Q000235:genetics
D001426:Bacterial Proteins; C096686:FemA protein, Bacteria; D012834:Silver; D006046:Gold; D004247:DNA