Cowpea mosaic virus nanoscaffold as signal enhancement for DNA microarrays.
Previous studies have shown that a functionalized viral nanoparticle can be used as a fluorescent signal-generating element and enhance detection sensitivity for immunoassays and low density microarrays. In this study, we further tested this ability in commercial DNA microarrays, including Affymetrix high density resequencing microarray. Optimum conditions for NeutrAvidin and dye coupling to a double-cysteine mutant of cowpea mosaic virus (CPMV) were found to be comparable to the commonly used streptavidin-phycoerythrin (SAPE) for high density resequencing microarray. A 3-fold signal enhancement in comparison to Cy5-dCTP controls was obtained when using nanoparticles on control scorecard expression microarrays. Hybridization results from commercially available 8000 rat expression arrays indicate an increment of 14% on the detected features when the virus complex was used as the staining reagent in comparison to Cy5-dCTP controls. The current work shows the utility of the CPMV-dye nanoparticles as a detection reagent in well-established detection platforms.
Soto
Carissa M
CM
Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA. cmsoto@cbmse.nrl.navy.mil
Blaney
Kate M
KM
Dar
Mubasher
M
Khan
Manzer
M
Lin
Baochuan
B
Malanoski
Anthony P
AP
Tidd
Cherise
C
Rios
Mayrim V
MV
Lopez
Darlah M
DM
Ratna
Banahalli R
BR
D000818:Animals; D001360:Avidin / Q000737:chemistry; D004396:Coloring Agents / Q000737:chemistry; D017798:Comovirus / Q000737:chemistry* / Q000235:genetics; D004247:DNA / Q000032:analysis*; D008958:Models, Molecular; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D051381:Rats; D012680:Sensitivity and Specificity
D004396:Coloring Agents; C093483:neutravidin; D001360:Avidin; D004247:DNA