WO2008047272A2 - Device for amplification and detection of nucleic acids - Google Patents

Device for amplification and detection of nucleic acids Download PDF

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Publication number
WO2008047272A2
WO2008047272A2 PCT/IB2007/054141 IB2007054141W WO2008047272A2 WO 2008047272 A2 WO2008047272 A2 WO 2008047272A2 IB 2007054141 W IB2007054141 W IB 2007054141W WO 2008047272 A2 WO2008047272 A2 WO 2008047272A2
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WO
WIPO (PCT)
Prior art keywords
compartment
nucleic acids
substrate
detection
cap
Prior art date
Application number
PCT/IB2007/054141
Other languages
French (fr)
Other versions
WO2008047272A3 (en
Inventor
Erik R. Vossenaar
Derk J. W. Klunder
Original Assignee
Koninklijke Philips Electronics N.V.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Koninklijke Philips Electronics N.V. filed Critical Koninklijke Philips Electronics N.V.
Priority to JP2009532925A priority Critical patent/JP2010506583A/en
Priority to US12/445,787 priority patent/US20100173794A1/en
Priority to EP07826707A priority patent/EP2081687A2/en
Priority to BRPI0717634-1A priority patent/BRPI0717634A2/en
Publication of WO2008047272A2 publication Critical patent/WO2008047272A2/en
Publication of WO2008047272A3 publication Critical patent/WO2008047272A3/en

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5082—Test tubes per se
    • B01L3/50825—Closing or opening means, corks, bungs
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • B01L3/50853—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates with covers or lids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/046—Function or devices integrated in the closure
    • B01L2300/047—Additional chamber, reservoir
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/00029—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor provided with flat sample substrates, e.g. slides
    • G01N2035/00099—Characterised by type of test elements
    • G01N2035/00158—Elements containing microarrays, i.e. "biochip"

Definitions

  • the invention relates to a device and a method for amplifying and detecting nucleic acid fragments, especially by hybridization.
  • PCR polymerase chain reaction
  • Detection of amplified PCR products may be done by hybridization to immobilized complementary nucleic acid sequences.
  • immobilized complementary nucleic acid sequences By immobilizing the specific complementary sequences (so-called capture probes) in a defined pattern, multiple nucleotide sequences can be detected simultaneously.
  • capture probes Such a patterned array of capture probes is often referred to as micro-array.
  • Nucleic sequences hybridized to the micro array can be detected by optical means, e.g. by fluorescence.
  • WO-A-01/45843 discloses a system for performing hybridization assays that comprises a cartridge for housing a flow-through device.
  • the flow through device has an array of microchannel passages.
  • the cartridge may include an observation window.
  • WO-A-00/12675 discloses a self-contained device which is described to be easily operated and which is devoid of contamination. The idea described provides for the rapid and accurate detection of amplified nucleic acids using a self-contained device.
  • the device integrates nucleic acid extraction, specific target amplification and detection into a single device, permitting rapid and accurate nucleic acid sequence detection.
  • the self contained device comprises a first hollow cylinder with a single closed end and a plurality of chambers therein, a second hollow elongated cylinder positioned contiguously inside the first cylinder capable of relative rotation. Sample is introduced in the first cylinder for extraction. The extracted nucleic acids is bound to a solid phase, and therefore not eluted from the solid phase by the addition of wash buffer. Amplification and labeling takes place in the same cylinder. Finally, the labeled, amplified product is reacted with microparticles conjugated with receptor specific ligands for the detection of the target sequence.
  • the invention therefore relates to a device for amplification and detection of nucleic acids, comprising a tube (1), which comprises at least one compartment (3) comprising reaction composition, and the tube comprising a cap (2) wherein the cap comprises a microarray of nucleic acids on its inside.
  • the invention further relates to a method for detecting real time hybridization of nucleic acids to a capture probe, which comprises the steps of: a) Administering a sample comprising PCR mastermix, appropriate PCR primers and template DNA to a bottom compartment of the device according to the invention, b) administering hybridization buffer to compartment (3) c) closing the device with the cap comprising a microarray of nucleic acids (capture probes) on its inside d) carrying out a PCR reaction for amplification of template DNA e) twisting the device such that the contents of compartment (3) flow out of the compartment f) optionally twisting the device repeatedly to ensure mixing of its contents with the contents of the device g) hybridization of the amplified template DNA with the capture probes h) detecting the hybridized amplified template DNA.
  • the invention relates to a system for amplification and detection of nucleic acids, comprising at least one, preferably a multitude of devices as described above.
  • Fig. 1 shows a device according to the invention.
  • Fig. 2 shows a system according to the invention.
  • Fig. 3 illustrates the method according to the invention. DETAILED DESCRIPTION
  • Template DNA is DNA that is present in a sample and of which the presence is to be detected.
  • PCR mastermix is a concentrated mix of PCR reaction components.
  • a mix comprises a composition selected from the group comprising polymerase enzyme, buffer, nucleotides or a combination thereof.
  • Microarray is defined as a set of miniaturized chemical reaction areas that are used to test nucleotide fragments, preferably DNA fragments.
  • the tube comprises a compartment (3) that comprises reagents that may be mixed with components present in the bottom compartment of tube (1) at a specific stage in the reaction, preferably after amplification of any target sequence present in a sample in the bottom of tube (1).
  • the at least one compartment (3) is preferably filled with reaction composition which is most preferred hybridization buffer in case of PCR reactions being carried out.
  • the compartment (3) preferably comprises an opening at the top side.
  • the compartment may be manipulated such that its contents are released into the tube (1).
  • Such manipulation may e.g. be by turning the device/tube 180 degrees.
  • the compartment (3) is made of a specific material which may be made permeable or may be ruptured by a specific trigger.
  • a specific trigger may be a temperature change, the application of light of a specific wavelength or a chemical reaction that is caused to take place in the tube (1).
  • the compartment (3) has an opening at its top side as specified above.
  • the device according to the invention is preferably integrated into a system comprising a heating element (4), a reader (5) and a transparent bottom plate (6).
  • Such a system for amplification and detection of nucleic acids preferably comprises at least one, more preferably a multitude of devices according to the invention.
  • the system preferably comprises a heating element (4), and a reader (5).
  • the system preferably further comprises a turning element that serves to rotate the device or devices.
  • Preferred readers include CCD camera's.
  • the system most preferably comprises a transparent bottom plate.
  • the invention in another aspect relates to a method for detecting real time hybridization of nucleic acids to a capture probe, which comprises the steps of: a) Administering a sample comprising PCR mastermix, appropriate PCR primers and template nucleic acid, preferably DNA to a bottom compartment of the device as described above, b) administering hybridization buffer to compartment (3) c) closing the device with the cap comprising a microarray of nucleic acids
  • capture probes on its inside d) carrying out a PCR reaction for amplification of template nucleic acid e) twisting the device such that the contents of compartment (3) flow out of the compartment f) optionally twisting the device repeatedly to ensure mixing of its contents with the contents of the device g) hybridizing the amplified template nucleic acid with the capture probes h) detecting the hybridized amplified template nucleic acid.
  • detection is preferably carried out using a scanning reader, most preferred a CCD camera.
  • Fig. 1 shows a schematic picture of a reaction tube.
  • a sample comprising PCR mastermix, appropriate (labelled) PCR primers and template DNA is administered to the bottom of the tube (1).
  • Hybridization buffer is administered to the open fluid compartment (3) inside of the tube (1).
  • the tube is capped with a cap (2) that comprises a microarray of nucleic acids on its inside.
  • the tube (1) is put into an integrated reader device (Fig. 2) which is herein referred to as the system (8).
  • This system comprises of a movable thermoblock capable of (rapid) thermocycling (heating element 4), a heated top lid and a transparent bottom plate (6). Below the bottom plate a confocal optical reader (5) is positioned.
  • the heating element may be used for temperature regulation in thermocycling, isothermal amplification or for sample heating during hybridization.
  • a sample is to be analysed, it is administered to the tube as described above and then put into the thermoblock (4) (Fig. 3.1).
  • This figure shows an example of a thermoblock capable of holding multiple tubes. A device for a single tube is also feasible.
  • the heating block (4) with the tubes is moved upward to ensure that the caps of the tubes are compressed to a heated lid (3.2). This lid it kept at a constant temperature slightly above 100 0 C to prevent sample evaporation during PCR thermocycling.
  • the thermoblock is moved down and twisted 180° (3.3). This causes the fluid to flow to the top of the tubes and the hybridization buffer to flow out of its compartment (3).
  • the block may be twisted repeatedly to ensure proper mixing of PCR fluid and hybridization buffer.
  • the block may be moved upright and up again for a 95°C denaturing step.
  • the amplified PCR products are allowed to hybridize to the capture probes on the micro array.
  • the block is moved down to a transparent plate.
  • a confocal reader capable of detecting fluorescence. Because the reader is confocal, the measurement (detection and excitation) volume is reduced substantially to typically a few micrometers away from the microarray surface, and thus the reader has enhanced surface specificity. As a result of the enhanced surface specificity, hybridization can be measured in real-time since no washing or removal of fluids is needed. This is illustrated in Fig. 3.4.
  • the reader can be a scanning reader.
  • evanescent excitation methods where an evanescent wave is excited at the surface of the substrate and excites surface bound fluorophores.
  • the surface of the cap positioned at the inner side of the cap, is the substrate.
  • TIR total internal reflection
  • a non- transparent composition such as a metal
  • the substrate may be covered with [an array of] apertures with at least one dimension in the plane parallel to the substrate-fluid interface below the diffraction limit of light in the fluid.
  • a non- transparent composition such as a metal
  • the substrate with [an array of] apertures with at least one dimension in the plane parallel to the substrate-fluid interface below the diffraction limit of light in the fluid.
  • wire grids that have one in-plane dimension above and the other dimension below the diffraction limit of the light in the fluid. This results in excitation volumes within 50 nm

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

A device is provided which is suitable for use in the amplification and detection of nucleic acids. The device comprises a sample tube comprising a compartment (3) that is preferably open at the top such that its content flows out when the tubed) is turned upside down. The tube further comprises a cap (2) which comprises a microarray of nucleic acids to which amplified sample DNA may hybridize.

Description

Device for amplification and detection of nucleic acids
FIELD OF THE INVENTION
The invention relates to a device and a method for amplifying and detecting nucleic acid fragments, especially by hybridization.
BACKGROUND TO THE INVENTION
PCR (polymerase chain reaction) is a method to amplify specific nucleic acid fragments. It is often used in research and for diagnosis of various diseases, including many infectious diseases. Detection of amplified PCR products may be done by hybridization to immobilized complementary nucleic acid sequences. By immobilizing the specific complementary sequences (so-called capture probes) in a defined pattern, multiple nucleotide sequences can be detected simultaneously. Such a patterned array of capture probes is often referred to as micro-array. Nucleic sequences hybridized to the micro array can be detected by optical means, e.g. by fluorescence.
WO-A-01/45843 discloses a system for performing hybridization assays that comprises a cartridge for housing a flow-through device. The flow through device has an array of microchannel passages. The cartridge may include an observation window. Although this system may be functional in a specialized laborotary environment, it does not fulfill the need for a simply operated device that may be operated at a point of care.
WO-A-00/12675 discloses a self-contained device which is described to be easily operated and which is devoid of contamination. The idea described provides for the rapid and accurate detection of amplified nucleic acids using a self-contained device. The device integrates nucleic acid extraction, specific target amplification and detection into a single device, permitting rapid and accurate nucleic acid sequence detection. The self contained device comprises a first hollow cylinder with a single closed end and a plurality of chambers therein, a second hollow elongated cylinder positioned contiguously inside the first cylinder capable of relative rotation. Sample is introduced in the first cylinder for extraction. The extracted nucleic acids is bound to a solid phase, and therefore not eluted from the solid phase by the addition of wash buffer. Amplification and labeling takes place in the same cylinder. Finally, the labeled, amplified product is reacted with microparticles conjugated with receptor specific ligands for the detection of the target sequence.
It is an object of the invention to provide an alternative simple system for amplifying and detecting nucleic acids.
SUMMARY OF THE INVENTION
The invention therefore relates to a device for amplification and detection of nucleic acids, comprising a tube (1), which comprises at least one compartment (3) comprising reaction composition, and the tube comprising a cap (2) wherein the cap comprises a microarray of nucleic acids on its inside.
The invention further relates to a method for detecting real time hybridization of nucleic acids to a capture probe, which comprises the steps of: a) Administering a sample comprising PCR mastermix, appropriate PCR primers and template DNA to a bottom compartment of the device according to the invention, b) administering hybridization buffer to compartment (3) c) closing the device with the cap comprising a microarray of nucleic acids (capture probes) on its inside d) carrying out a PCR reaction for amplification of template DNA e) twisting the device such that the contents of compartment (3) flow out of the compartment f) optionally twisting the device repeatedly to ensure mixing of its contents with the contents of the device g) hybridization of the amplified template DNA with the capture probes h) detecting the hybridized amplified template DNA.
In a further instance the invention relates to a system for amplification and detection of nucleic acids, comprising at least one, preferably a multitude of devices as described above.
DESCRIPTION OF THE FIGURES:
Fig. 1 shows a device according to the invention. Fig. 2 shows a system according to the invention. Fig. 3 illustrates the method according to the invention. DETAILED DESCRIPTION
Definitions
Template DNA is DNA that is present in a sample and of which the presence is to be detected.
PCR mastermix is a concentrated mix of PCR reaction components. Generally such a mix comprises a composition selected from the group comprising polymerase enzyme, buffer, nucleotides or a combination thereof.
Microarray is defined as a set of miniaturized chemical reaction areas that are used to test nucleotide fragments, preferably DNA fragments.
The tube comprises a compartment (3) that comprises reagents that may be mixed with components present in the bottom compartment of tube (1) at a specific stage in the reaction, preferably after amplification of any target sequence present in a sample in the bottom of tube (1). The at least one compartment (3) is preferably filled with reaction composition which is most preferred hybridization buffer in case of PCR reactions being carried out.
The compartment (3) preferably comprises an opening at the top side. In this way, the compartment may be manipulated such that its contents are released into the tube (1). Such manipulation may e.g. be by turning the device/tube 180 degrees.
Alternatively the compartment (3) is made of a specific material which may be made permeable or may be ruptured by a specific trigger. Such trigger may be a temperature change, the application of light of a specific wavelength or a chemical reaction that is caused to take place in the tube (1). Most preferred the compartment (3) has an opening at its top side as specified above.
The device according to the invention is preferably integrated into a system comprising a heating element (4), a reader (5) and a transparent bottom plate (6).
Such a system for amplification and detection of nucleic acids, preferably comprises at least one, more preferably a multitude of devices according to the invention. For carrying out a PCR reaction the system preferably comprises a heating element (4), and a reader (5).
The system preferably further comprises a turning element that serves to rotate the device or devices. Preferred readers include CCD camera's.
To facilitate the detection, the system most preferably comprises a transparent bottom plate.
In another aspect the invention relates to a method for detecting real time hybridization of nucleic acids to a capture probe, which comprises the steps of: a) Administering a sample comprising PCR mastermix, appropriate PCR primers and template nucleic acid, preferably DNA to a bottom compartment of the device as described above, b) administering hybridization buffer to compartment (3) c) closing the device with the cap comprising a microarray of nucleic acids
(capture probes) on its inside d) carrying out a PCR reaction for amplification of template nucleic acid e) twisting the device such that the contents of compartment (3) flow out of the compartment f) optionally twisting the device repeatedly to ensure mixing of its contents with the contents of the device g) hybridizing the amplified template nucleic acid with the capture probes h) detecting the hybridized amplified template nucleic acid. In this method detection is preferably carried out using a scanning reader, most preferred a CCD camera.
The invention is illustrated by a first embodiment as described below. Fig. 1 shows a schematic picture of a reaction tube. A sample, comprising PCR mastermix, appropriate (labelled) PCR primers and template DNA is administered to the bottom of the tube (1). Hybridization buffer is administered to the open fluid compartment (3) inside of the tube (1). The tube is capped with a cap (2) that comprises a microarray of nucleic acids on its inside.
The tube (1) is put into an integrated reader device (Fig. 2) which is herein referred to as the system (8). This system comprises of a movable thermoblock capable of (rapid) thermocycling (heating element 4), a heated top lid and a transparent bottom plate (6). Below the bottom plate a confocal optical reader (5) is positioned. The heating element may be used for temperature regulation in thermocycling, isothermal amplification or for sample heating during hybridization. When a sample is to be analysed, it is administered to the tube as described above and then put into the thermoblock (4) (Fig. 3.1). This figure shows an example of a thermoblock capable of holding multiple tubes. A device for a single tube is also feasible. The heating block (4) with the tubes is moved upward to ensure that the caps of the tubes are compressed to a heated lid (3.2). This lid it kept at a constant temperature slightly above 1000C to prevent sample evaporation during PCR thermocycling. After thermocycling is complete, the thermoblock is moved down and twisted 180° (3.3). This causes the fluid to flow to the top of the tubes and the hybridization buffer to flow out of its compartment (3). Optionally, the block may be twisted repeatedly to ensure proper mixing of PCR fluid and hybridization buffer. Optionally, the block may be moved upright and up again for a 95°C denaturing step. When the tubes are upside down, the amplified PCR products are allowed to hybridize to the capture probes on the micro array. The block is moved down to a transparent plate. Below this plate is a confocal reader capable of detecting fluorescence. Because the reader is confocal, the measurement (detection and excitation) volume is reduced substantially to typically a few micrometers away from the microarray surface, and thus the reader has enhanced surface specificity. As a result of the enhanced surface specificity, hybridization can be measured in real-time since no washing or removal of fluids is needed. This is illustrated in Fig. 3.4. The reader can be a scanning reader.
As an alternative method for enhancing the surface specificity, one can use evanescent excitation methods, where an evanescent wave is excited at the surface of the substrate and excites surface bound fluorophores. In this context the surface of the cap, positioned at the inner side of the cap, is the substrate.
As a first method one can use total internal reflection (TIR) at the substrate/ fluid interface, which results in measurement (excitation) volumes within 100-500 nm of the array surface. TIR however prefers the use of a glass prism connected to the substrate or the use of a substrate with a wedge shape to enable the coupling of excitation light with angles above the critical angle of the substrate-fluid interface into the substrate.
As a second preferred method one may cover the substrate with a non- transparent composition such as a metal and pattern the substrate with [an array of] apertures with at least one dimension in the plane parallel to the substrate-fluid interface below the diffraction limit of light in the fluid. As an example, one can pattern the substrate with wire grids that have one in-plane dimension above and the other dimension below the diffraction limit of the light in the fluid. This results in excitation volumes within 50 nm

Claims

(typically 20-30 nm) of the array surface. Advantage of this method over the first method [for evanescent excitation] is that it is simpler- there is no need for a prism or wedge shaped surface and that there are no special requirements for the angle of incidence and shape of the excitation spot and one can use a simple CCD camera for imaging the fluorescence- and enables substantially smaller excitation volumes.In the embodiment where the substrate is patterened with a wire grid, the array of nucleotides can ideally be positioned in the openings of the wire grid.For further detection techniques reference is for example made to international application IB2005/053168 which is incorporated by reference. CLAIMS:
1. A device for amplification and detection of nucleic acids, comprising a tube (1), which comprises at least one compartment (3) comprising reaction composition, and the tube comprising a cap (2) wherein the cap comprises a microarray of nucleic acids on its inside.
2. Device according to claim 1 wherein the at least one compartment (3) is filled with reaction composition which is hybridization buffer.
3. Device according to any of claims 1-2 wherein the compartment (3) may be manipulated such that its contents are released into the tube (1).
4. Device according to any of claims 1-3 wherein the compartment (3) comprises an opening at its top side.
5. Device according to any of claims 1-4 which is integrated into a system comprising a heating element (4), a reader (5) and a transparent bottom plate (6).
6. System for amplification and detection of nucleic acids, comprising at least one, preferably a multitude of devices according to any of claims 1-4.
7. System according to claim 6 further comprising, a heating element (4), and a reader (5).
8. System according to claim 6, further comprising a turning element that serves to rotate the device or devices.
9. System according to claim 6, wherein the reader is a confocal reader.
10. System according to claim 6, comprising a surface of the cap as substrate, wherein the substrate is wedge shaped in order to enable excitation of fluorophores at the microarray surface.
11. System according to claim 6, comprising a surface of the cap as substrate wherein the substrate is attached to a prism in order to enable excitation of fluorophores at the array surface.
12. System according to claim 6, comprising a suface of the cap as substrate , where the substrate is covered at the interface facing the fluid with a patterned composition in order to enable evanescent excitation at fluorophores at the array surface.
13. System according to claim 6 further comprising a transparent bottom plate.
14. Method for detecting real time hybridization of nucleic acids to a capture probe, which comprises the steps of: a) Administering a sample comprising PCR mastermix, appropriate PCR primers and template nucleic acids, preferably DNA, to a bottom compartment of the device according to claim 1 , b) administering hybridization buffer to compartment (3) c) closing the device with the cap comprising a microarray of nucleic acids, called capture probes, on its inside d) carrying out a PCR reaction for amplification of template nucleic acid e) twisting the device such that the contents of compartment (3) flow out of the compartment f) optionally twisting the device repeatedly to ensure mixing of its contents with the contents of the device g) hybridizing the amplified template nucleic acid with the capture probes h) detecting the hybridized amplified template nucleic acid.
15. Method according to claim 14 wherein detection is carried out using a scanning reader.
16. Method according to claim 14 for use in the detection of infectious diseases.
PCT/IB2007/054141 2006-10-17 2007-10-11 Device for amplification and detection of nucleic acids WO2008047272A2 (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
JP2009532925A JP2010506583A (en) 2006-10-17 2007-10-11 Nucleic acid amplification and detection device
US12/445,787 US20100173794A1 (en) 2006-10-17 2007-10-11 Device for amplification and detection of nucleic acids
EP07826707A EP2081687A2 (en) 2006-10-17 2007-10-11 Device for amplification and detection of nucleic acids
BRPI0717634-1A BRPI0717634A2 (en) 2006-10-17 2007-10-11 NUCLEIC ACID AMPLIFICATION AND DETECTION DEVICE, NUCLEIC ACID AMPLIFICATION AND DETECTION SYSTEM, AND REAL-TIME NUCLEIC ACID HYBRIDIZATION METHOD FOR A CATCH PROBE

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP06122465 2006-10-17
EP06122465.5 2006-10-17

Publications (2)

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WO2008047272A2 true WO2008047272A2 (en) 2008-04-24
WO2008047272A3 WO2008047272A3 (en) 2008-06-12

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EP (1) EP2081687A2 (en)
JP (1) JP2010506583A (en)
CN (1) CN101528351A (en)
BR (1) BRPI0717634A2 (en)
RU (1) RU2009118455A (en)
WO (1) WO2008047272A2 (en)

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WO2017127570A1 (en) * 2016-01-20 2017-07-27 Triv Tech, Llc Point-of-care nucleic acid amplification and detection
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