scholarly journals Silicon-based Biosensors for Rapid Detection of Protein or Nucleic Acid Targets

2001 ◽  
Vol 47 (10) ◽  
pp. 1894-1900 ◽  
Author(s):  
Robert Jenison ◽  
Helen La ◽  
Ayla Haeberli ◽  
Rachel Ostroff ◽  
Barry Polisky

Abstract Background: We developed a silicon-based biosensor that generates visual, qualitative results or quantitative results for the detection of protein or nucleic acid targets in a multiplex format. Methods: Capture probes were immobilized either passively or covalently on the optically coated surface of the biosensor. Intermolecular interactions of the immobilized capture probe with specific target molecules were transduced into a molecular thin film. Thin films were generated by enzyme-catalyzed deposition in the vicinity of the surface-bound target. The increased thickness on the surface changed the apparent color of the biosensor by altering the interference pattern of reflected light. Results: Cytokine detection was achieved in a 40-min multiplex assay. Detection limits were 4 ng/L for interleukin (IL)-6, 31 ng/L for IL1-β, and 437 ng/L for interferon-γ. In multianalyte experiments, cytokines were specifically detected with signal-to-noise ratios ranging from 15 to 80. With a modified optical surface, specificity was also demonstrated in a nucleic acid array with unambiguous discrimination of single-base changes in a 15-min assay. For homozygous wild-type and homozygous mutant samples, signal-to-noise ratios of ∼100 were observed. Heterozygous samples yielded approximately equivalent signals for wild-type and mutant capture probes. Conclusions: The thin-film biosensor allows rapid, sensitive, and specific detection of protein or nucleic acid targets in an array format with results read visually or quantified with a charge-coupled device camera. This biosensor is suited for multianalyte detection in clinical diagnostic assays.

1999 ◽  
Vol 45 (9) ◽  
pp. 1659-1664 ◽  
Author(s):  
Rachel M Ostroff ◽  
Deborah Hopkins ◽  
Ayla B Haeberli ◽  
Wahab Baouchi ◽  
Barry Polisky

Abstract Background: We have developed a silicon-based biosensor that generates a visual signal in response to nucleic acid targets. Methods: In this system, capture oligonucleotide probes are immobilized on the surface of the biosensor. Interaction of the capture probes with a complementary target and a biotinylated detector oligonucleotide allows initiation of formation of an organic thin film on the biosensor. Thin film formation is completed by enzymatic activity of peroxidase conjugated to an anti-biotin antibody. Peroxidase catalyzes deposition of an insoluble product onto the silicon surface, generating a uniform thin film. The increased thickness on the surface alters the perceived color of the biosensor through changes in the interference patterns of reflected light from the surface, causing a color change from gold to purple. Results: The biosensor results may be evaluated by direct visual inspection or quantified by ellipsometry. Results are obtained in 25 min with a detection limit of 5 pmol/L (150 amol/sample). Selectivity of the biosensor is demonstrated by discrimination of single nucleotide mismatches. Multitarget arrays are also analyzed with the thin film biosensor, and the system is capable of detecting targets from human serum and urine. Conclusions: The biosensor surface is inexpensive to produce, and the assay format is simple and rapid. The thin film biosensor is adaptable to a wide variety of nucleic acid detection applications, including rapid diagnostic testing for infectious disease panels, antibiotic resistance panels, or allelic discrimination of specific genetic markers.


Nanophotonics ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 3443-3450 ◽  
Author(s):  
Wei-Nan Liu ◽  
Rui Chen ◽  
Wei-Yi Shi ◽  
Ke-Bo Zeng ◽  
Fu-Li Zhao ◽  
...  

AbstractSelective transmission or filtering always responds to either frequency or incident angle, so as hardly to maximize signal-to-noise ratio in communication, detection and sensing. Here, we propose compact meta-filters of narrow-frequency sharp-angular transmission peak along with broad omnidirectional reflection sidebands, in all-dielectric cascaded subwavelength meta-gratings. The inherent collective resonance of waveguide-array modes and thin film approximation of meta-grating are employed as the design strategy. A unity transmission peak, locating at the incident angle of 44.4° and the center wavelength of 1550 nm, is demonstrated in a silicon meta-filter consisting of two-layer silicon rectangular meta-grating. These findings provide possibilities in cascaded meta-gratings spectroscopic design and alternative utilities for high signal-to-noise ratio applications in focus-free spatial filtering and anti-noise systems in telecommunications.


2006 ◽  
Vol 52 (10) ◽  
pp. 1855-1863 ◽  
Author(s):  
Giulia Amicarelli ◽  
Daniel Adlerstein ◽  
Erlet Shehi ◽  
Fengfei Wang ◽  
G Mike Makrigiorgos

Abstract Background: Genotyping methods that reveal single-nucleotide differences are useful for a wide range of applications. We used digestion of 3-way DNA junctions in a novel technology, OneCutEventAmplificatioN (OCEAN) that allows sequence-specific signal generation and amplification. We combined OCEAN with peptide-nucleic-acid (PNA)-based variant enrichment to detect and simultaneously genotype v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) codon 12 sequence variants in human tissue specimens. Materials and Methods: We analyzed KRAS codon 12 sequence variants in 106 lung cancer surgical specimens. We conducted a PNA-PCR reaction that suppresses wild-type KRAS amplification and genotyped the product with a set of OCEAN reactions carried out in fluorescence microplate format. The isothermal OCEAN assay enabled a 3-way DNA junction to form between the specific target nucleic acid, a fluorescently labeled “amplifier”, and an “anchor”. The amplifier-anchor contact contains the recognition site for a restriction enzyme. Digestion produces a cleaved amplifier and generation of a fluorescent signal. The cleaved amplifier dissociates from the 3-way DNA junction, allowing a new amplifier to bind and propagate the reaction. Results: The system detected and genotyped KRAS sequence variants down to ∼0.3% variant-to-wild-type alleles. PNA-PCR/OCEAN had a concordance rate with PNA-PCR/sequencing of 93% to 98%, depending on the exact implementation. Concordance rate with restriction endonuclease-mediated selective-PCR/sequencing was 89%. Conclusion: OCEAN is a practical and low-cost novel technology for sequence-specific signal generation. Reliable analysis of KRAS sequence alterations in human specimens circumvents the requirement for sequencing. Application is expected in genotyping KRAS codon 12 sequence variants in surgical specimens or in bodily fluids, as well as single-base variations and sequence alterations in other genes.


Author(s):  
D. N. Redka ◽  
V.A. Parfenov ◽  
V.P. Afanasjev ◽  
A.V. Kukin ◽  
F. S. Egorov ◽  
...  

2001 ◽  
Vol 685 ◽  
Author(s):  
M. Fernandes ◽  
Yu. Vygranenko ◽  
J. Martins ◽  
M. Vieira

AbstractWe suggest to enhance the performance of image acquisition systems based on large area amorphous silicon based sensors by optimizing the readout parameters such as the intensity and cross-section of scanner beam, acquisition time and bias conditions. The main output device characteristics as image responsivity, signal to noise ratio and spatial resolution were analyzed in open circuit, short circuit and photodiode modes. The result show that the highest signal to noise ratio and best dark to bright ratio can be achieved in short circuit mode.It was shown that the sensor resolution is related to the basic device parameters and, in practice, limited by the acquisition time and scanning beam properties. The scanning beam spot size limits the resolution due to the overlapping of dark and illuminated zones leading to a blurring effect on the final image and a consequent degradation in the resolution.


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