Measurement of Analyte with an Acoustic Wave Sensor

Inactive Publication Date: 2018-07-12
FOUND FOR RES & TECH HELLAS
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  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0045]The analyte may be dsDNA fragments produced by any type of enzymatic amplification (PCR or isothermal) or hybridization or any other enzymatic reaction. In this case, the analyte may comprise one or more binding moieties to form bonds (typically covalent bonds) with the sensing surface and/or the label body. Direct binding of the ds analyte to the sensor surface, for example through a thiol modification, may eliminate the need for a specific surface-bound recognition molecule. If the analyte is sufficiently long, it may eliminate the benefit of a separate spacer region.

Problems solved by technology

Still, today, while PCR represents the ultimate in terms of sensitivity, it has significant drawbacks including complexity, sensitivity to contamination, cost and lack of portability (Rosi, N. L.; Mirkin, C. A.; Nanostructures in Biodiagnostics, Chem. Rev. 105:1547-1562 (2005)).
In some settings, PCR bias can cause 10 to 30-fold differences in amplification efficiency which could result in underestimation or failure to detect mutations.
In the case of heterogeneous samples where rare mutated sequences exist amongst abundant wild-type sequences, the PCR may be unable to amplify sufficiently these rare targets.
Furthermore, non-specific PCR inhibitors, including heparin, and uncharacterized components are sometimes present in samples from patients which may lead to undesired results such as mis-priming and inhibition.
However, this impressive performance, similar to that obtained with PCR, does not come in a simple format; it involves cumbersome and lengthy procedures such as the use of exogenous surface-modified components and multi-step amplification and detection schemes.
However, the mechanism by which acoustic energy is dissipated when biomolecules are attached to the device surface is still unclear and unexploited in clinical applications.
Briefly, a drag force is produced by oscillating biomolecules (attached to the surface via a single point) in the surrounding liquid and this is energy consuming.

Method used

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  • Measurement of Analyte with an Acoustic Wave Sensor
  • Measurement of Analyte with an Acoustic Wave Sensor
  • Measurement of Analyte with an Acoustic Wave Sensor

Examples

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Example Implementation

[0082]In an example implementation for the detection of a single stranded nucleic acid analyte the acoustic wave sensor is a Quartz Crystal Microbalance (QCM) constructed as described in the Materials and Methods section below. The sensor has a quartz crystal substrate 2 and a sensing surface formed by a surface gold layer 4 to which neutravidin is adsorbed. A 5′-biotinylated single stranded DNA molecule is used as the surface probe 6. This probe is formed through PCR or an isothermal amplification process using a suitable set of primers and introduced to the liquid medium which is in contact with the sensing surface. (Short 5′-biotinylated single stranded DNA molecules are also commercially available). The surface probe single stranded DNA adheres to the sensing surface by virtue of the specific interaction between biotin 8 and neutravidin on the sensing surface. Each DNA molecule is individually attached through the biotin which is attached to the end of the DNA

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Abstract

A sensitive assay for an analyte employing an acoustic wave sensor. A label which has a higher dissipative capacity than the analyte is adhered to the sensing surface of an acoustic wave sensor through the analyte such that the body of the label is spaced apart from and anchored to the surface of the acoustic wave sensor by a distance of 15 to 250 nm. The change in the energy losses of the acoustic wave when the label binds to the sensing surface is used to measure the presence or amount of the label. A substantial improvement in the detection limit of the label is obtained. The analyte may for example be a nucleic acid and the label may for example comprise liposomes.

Description

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Claims

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Application Information

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Owner FOUND FOR RES & TECH HELLAS
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