Gold nanoparticle-based detection of genomic DNA targets on microarrays using a novel optical detection system.
The development of a nanoparticle-based detection methodology for sensitive and specific DNA-based diagnostic applications is described. The technology utilizes gold nanoparticles derivatized with thiol modified oligonucleotides that are designed to bind complementary DNA targets. A glass surface with arrays of immobilized oligonucleotide capture sequences is used to capture DNA targets, which are then detected via hybridization to the gold nanoparticle probes. Amplification with silver allows for detection and quantitation by measuring evanescent wave induced light scatter with low-cost optical detection systems. Compared to Cy3-based fluorescence, silver amplified gold nanoparticle probes provide for a approximately 1000-fold increase in sensitivity. Furthermore, direct detection of non-amplified genomic DNA from infectious agents is afforded through increased specificity and even identification of single nucleotide polymorphisms (SNP) in human genomic DNA appears feasible.
Storhoff
James J
JJ
Nanosphere, Inc., Applied Research, 4088 Commercial Ave., Northbrook, IL 60062, USA. jstorhoff@nanosphere.us
Marla
Sudhakar S
SS
Bao
Paul
P
Hagenow
Susan
S
Mehta
Hitesh
H
Lucas
Adam
A
Garimella
Viswanadham
V
Patno
Tim
T
Buckingham
Wes
W
Cork
William
W
Müller
Uwe R
UR
D001426:Bacterial Proteins / Q000235:genetics; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000648:ultrastructure; D015342:DNA Probes / Q000737:chemistry*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods*; D043942:Nanotubes / Q000737:chemistry* / Q000648:ultrastructure*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D046915:Penicillin-Binding Proteins; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012834:Silver / Q000737:chemistry; D013194:Staining and Labeling; D013211:Staphylococcus aureus / Q000235:genetics
D001426:Bacterial Proteins; D015342:DNA Probes; D046915:Penicillin-Binding Proteins; C487789:mecA protein, Staphylococcus aureus; D012834:Silver; D006046:Gold; D004247:DNA