Interference-based detection of nucleic acid targets on optically coated silicon.
Sequence-specific detection of polynucleotides typically requires modified reporter probes that are labeled with radioactive, fluorescent, or luminescent moieties. Although these detection methods are capable of high sensitivity, they require instrumentation for signal detection. In certain settings, such as clinical point of care, instrumentation might be impractical or unavailable. Here we describe a detection approach in which formation of a nucleic acid hybrid is enzymatically transduced into a molecular thin film that can be visually detected in white light. The system exploits a flat, optically coated silicon-based surface to which capture oligonucleotides are covalently attached. The optimized system is capable of detection of nucleic acid targets present at sub-attomole levels. To supplement visual detection, signals can be quantitated by a charge-coupled device. The design and composition of the optical surface, optimization of immobilization chemistry for attachment of capture probes, and characterization of the efficiency of the hybridization process are presented. We describe the application of this system to detection of a clinically relevant target, the mecA gene present in methicillin-resistant Staphylococcus aureus.
Jenison
R
R
BioStar, Inc., 6655 Lookout Road, Boulder, CO 80301, USA.
Yang
S
S
Haeberli
A
A
Polisky
B
B
D001483:Base Sequence; D015374:Biosensing Techniques; D003432:Cross-Linking Reagents; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D006801:Humans; D016106:Methicillin Resistance / Q000235:genetics*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D015345:Oligonucleotide Probes / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D017655:Silicon Compounds; D013211:Staphylococcus aureus / Q000235:genetics*; D013499:Surface Properties
D003432:Cross-Linking Reagents; D004269:DNA, Bacterial; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D017655:Silicon Compounds; C032734:silicon nitride