Interference-based detection of nucleic acid targets on optically coated silicon

• Published in: Nature biotechnology Vol. 19; no. 1; pp. 62 - 65
• Main Authors: Polisky, Barry, Jenison, Robert, Yang, Shao, Haeberli, Ayla
• Format: Journal Article
• Language: English
• Published: New York, NY Nature 01.01.2001; Nature Publishing Group
• Subjects: Acids; Antibiotics; Base Sequence; Biological and medical sciences; Biosensing Techniques; Biosensors; Biotechnology; Cross-Linking Reagents; Diverse techniques; DNA, Bacterial - genetics; Drug resistance; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Humans; Hybridization; Instrumentation; Light; Methicillin Resistance - genetics; Methods. Procedures. Technologies; Molecular and cellular biology; Nucleic acids; Oligodeoxyribonucleotides - chemistry; Oligonucleotide Probes - chemistry; Probes; Publishing; Sequence Analysis, DNA; Silicon; Silicon Compounds; Silicon nitride; Staphylococcus aureus - genetics; Staphylococcus infections; Surface Properties; Thin films; Various methods and equipments; Molecular hybridization; DNA chip; Immobilization; Resistance; Antibiotic; Biosensor; Bacteria; Micrococcales; Micrococcaceae; Oligonucleotide; Silicon; Molecular probe; Diagnosis; Detection; Nucleic acid; Staphylococcus aureus
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/83530

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.