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WO2003106031A1 - Procede et dispositif de detection de la presence d'un analyte dans un echantillon pour essai - Google Patents

Procede et dispositif de detection de la presence d'un analyte dans un echantillon pour essai Download PDF

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Publication number
WO2003106031A1
WO2003106031A1 PCT/EP2003/005972 EP0305972W WO03106031A1 WO 2003106031 A1 WO2003106031 A1 WO 2003106031A1 EP 0305972 W EP0305972 W EP 0305972W WO 03106031 A1 WO03106031 A1 WO 03106031A1
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WO
WIPO (PCT)
Prior art keywords
chamber
test sample
analytical device
binding surface
nucleic acids
Prior art date
Application number
PCT/EP2003/005972
Other languages
English (en)
Inventor
Martin Kopp
Original Assignee
F. Hoffmann-La Roche Ag
Roche Diagnostics Gmbh
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 F. Hoffmann-La Roche Ag, Roche Diagnostics Gmbh filed Critical F. Hoffmann-La Roche Ag
Publication of WO2003106031A1 publication Critical patent/WO2003106031A1/fr

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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
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • 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
    • B01L2300/0654—Lenses; Optical fibres
    • 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
    • B01L2300/0681—Filter
    • 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
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0877—Flow chambers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/08—Regulating or influencing the flow resistance
    • B01L2400/084—Passive control of flow resistance
    • B01L2400/086—Passive control of flow resistance using baffles or other fixed flow obstructions
    • 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

Definitions

  • the present invention concerns a method, an analytical device and a system for detecting the presence or the amount of an analyte in a test sample.
  • An aim of the present invention is to provide a method, an analytical device and a system which are suitable for carrying out all above mentioned steps fully automatically without any transfer of the nucleic acid between the analyzing steps, thus reducing the- risk of sample contamination and loss of sample material. Furthermore the invention provides an analytical device with a simple structure, which therefore is particularly well suited for miniaturization and manufacturing at low costs.
  • a method according to the invention is defined by claim 1.
  • An analytical device according to the invention is defined by claim 7.
  • a system according to the invention is defined by claim 21.
  • the sample processing time is substantially reduced, since an elution step is not necessary and no transfer steps are involved .
  • analyte refers to a sequence of nucleic acid, e. g. DNA or RNA, whose presence in a test sample is to be detected.
  • Fig. 1 shows a schematic representation of an analytical device according to the invention
  • Fig. 2 shows a first embodiment of an analytical cell which can be used as part of the device shown in Fig. 1;
  • Fig. 3 shows a second embodiment of an analytical cell which can be used as part of the device shown in Fig. 1;
  • Fig. 4 shows an electron-microscopic picture of a fleece used as a binding surface in the analytical cell of Figs . 2 and 3 ;
  • Fig. 5 shows schematically the flow behavior in an analytical cell of the type shown in Fig. 2;
  • Fig. 6 shows schematically the flow behavior in an analytical cell of the type shown in Fig. 3 ;
  • Fig. 7 shows a schematic side view of an analytical cell of the type shown in Figs . 2 or 3 ;
  • Fig. 8 shows the results obtained by the analytical device shown in Fig. 2.
  • Fig. 9 shows a top view of an analytical cell of the type shown in Fig . 3 ,
  • Fig. 10 shows a magnified view of a portion of the surface of the analytical cell shown in Fig. 9, and
  • Fig. 11 shows a scanning-electron-microscopic picture of a portion of the surface of the analytical cell shown in Fig. 9.
  • Fig. 12 shows an embodiment of an analytical system according to the invention.
  • the present invention provides a method of detecting an amplified target nucleic acid sequence that is present in a sample. It is recognized by those skilled in the art that assays for a broad range of target nucleic acid sequences present in a sample may be performed in accordance with the present invention.
  • Samples may include biological samples derived from agriculture sources, bacterial and viral sources, and from human or other animal sources, as well as other samples such as waste or drinking water, agricultural products, processed foodstuff, air, etc. Examples include blood, stool, sputum, mucus, serum, urine, saliva, teardrop, a biopsy sample, an histological tissue sample, a tissue culture product, an agricultural product, waste or drinking water, foodstuff, air, etc.
  • the present invention is useful for the detection of nucleic acid sequences indicative of genetic defects or contagious diseases.
  • the following definitions will be helpful in understanding the specification and claims. The definitions provided herein should be borne in mind when these terms are used in the following examples and throughout the instant application.
  • amplification refers to a "template-dependent process” that results in an increase in the concentration of a nucleic acid sequence relative to its initial concentration.
  • a “template-dependent process” is defined as a process that involves the “template-dependent extension” of a “primer” molecule.
  • a “primer” molecule refers to a sequence of nucleic acid that is complementary to a portion of the target or control sequence and may or may not be labeled with a hapten.
  • a “template dependent extension” refers to nucleic acid synthesis of RNA or DNA wherein the sequence of the newly synthesized strand of nucleic acid is dictated by the rules of complementary base pairing of the target nucleic acid and the primers.
  • FIG. 1 An analytical device according to the invention for determining the presence of an analyte in a test sample is schematically represented in Fig. 1.
  • Analytical cell 11 comprises a chamber 12 and has a sealable inlet port 13 and a sealable outlet port 14.
  • the analytical device comprises a pre-treatment container 21 adapted to be connected to chamber 12 of analytical cell 11 through inlet port 11, and a waste container 22 adapted to be connected to chamber 12 of analytical cell 11 through outlet port 14 of analytical cell 11.
  • Container 21 is adapted for mixing a test sample with reagents, and for preincubation for lysing microbiological particles in order to prepare the nucleic acids in the test sample to be transferred to chamber 12 of analytical cell 11 via inlet port 13.
  • Container 21 is disconnectable from chamber 12 by sealing inlet port 13.
  • Container 21 has several inlets: inlet 23 for receiving a liquid sample, inlet 24 for receiving a reagent, inlet 25 for receiving air. In operation container 21 is connected through these inlets to corresponding sources of liquid respectively air under pressure.
  • Waste container 22 is adapted for receiving through outlet port 14 waste generated by process steps carried out within chamber 12 of analytical cell 11.
  • Container 22 is disconnectable from chamber 12 by sealing outlet port 14.
  • Fig. 2 shows a schematic representation of a first embodiment of analytical cell 11.
  • Chamber 12 of cell 11 has an inlet port 33 and outlet port 34. In Fig. 2 these ports are open and allow flow of liquid through chamber 12.
  • FIG. 3 shows a schematic representation of a second embodiment of cell 11.
  • Chamber 12a of cell 11 has an inlet port 33a and outlet port 34a. In Fig. 3 these ports are open and allow flow of liquid through chamber 12a.
  • This chamber 12 has wedge shaped inlet and outlet ports.
  • Fig. 5 shows schematically the flow behavior in an analytical cell of the type shown in Fig. 2.
  • Fig. 6 shows schematically the flow behavior in an analytical cell of the type shown in Fig. 3.
  • Fig. 7 shows a schematic side view of an analytical cell of the type shown in Figs. 2 or 3.
  • the upper part body of analytical cell 11 shown by Fig. 7 is preferably made of suitable plastic material e.g. of a transparent polycarbonate. Chamber 12 and inlet and outlet ports 33 respectively 34 are formed within this body. The bottom of cell 11 is an aluminum wall 16. Liquid flow through inlet 33 and through outlet 34 is represented by arrows. Wall
  • a thermal instrument e.g. a thermal cycler of the kind used to perform a polymerase chain reaction.
  • a thermal instrument e.g. a thermal cycler of the kind used to perform a polymerase chain reaction.
  • FIG. 7 there is an air gap 26 within the body of analytical cell.
  • a part of chamber 12 has a zone
  • arrow 19 represents an excitation light beam which irradiates body 15 located in chamber 12, and arrow 20 represents a fluorescent light beam emitted by reaction products in chamber 12.
  • Inlet port 33 and outlet port 34 allow flow of liquid through them.
  • Inlet port 33 allows introduction of liquid containing a sample to be tested into chamber 12.
  • Outlet port 34 of chamber 12 allows liquid to flow out of chamber 12.
  • inlet port 33a is wedge shaped and has a decreasing cross-section in the sense of liquid flow.
  • outlet port 34a is wedge shaped and has an increasing cross-section in the sense of liquid flow.
  • Chamber 12 of analytical cell 11 contains a binding surface which is adapted to capture nucleic acids contained in a test sample when the liquid containing the test sample flows through chamber 12.
  • the binding surface is part of a body 15 which is manufactured separately from the other parts of the analytical cell 11 and which has a large surface-to- volume ratio.
  • a body 15 can e.g. consist of beads, membranes, gels or fibers forming a fleece.
  • Fig. 4 shows an electron-microscopic picture of a glass fiber fleece which has a surface suitable as a binding surface in the analytical cells 11 and 11a shown by Figs. 2 and 3 respectively.
  • a preferred glass fiber fleece for making a suitable binding surface has e.g. the following composition:
  • Ti 1.11 2.50 SiO x and A1 2 0 3 are particularly important components of the fleece with regard to its capacity to bind nucleic acids.
  • a fleece consisting of pure Si0 2 shows an acceptable capacity to bind nucleic acids.
  • chamber 12 of analytical device 11 contains a binding surface which consists of plain fibers, e.g. a glass fiber fleece or a cellulose fleece.
  • the binding surface consists e.g. of glass or cellulose fibers which are coated with a lysing agent, i.e. a material which promotes the lysing of cells which contain the analyte and which enables the analyte to bind to said fibrous material .
  • a lysing agent i.e. a material which promotes the lysing of cells which contain the analyte and which enables the analyte to bind to said fibrous material .
  • the lysing agent coating is applied to the fibers before these are is introduced into chamber 12.
  • Preferred examples of material for a fleece coated for both purposes are commercially available materials (fleeces) like Schleicher and Schuell GmbH Isocode filter paper or grade 903 paper from the same supplier. Other materials serving the same purpose are the so called Whatman FTA (Trade Mark) cards. All these materials instantaneously lyse cells and subsequently bind the DNA or the RNA.
  • Fig. 12 shows another preferred embodiment of analytical cell 11.
  • the central part of the inner surface of the upper wall chamber 12 is manufactured by a micromachining process which yields a microstructured cell which has a high surface-to-volume ratio and which is a binding surface for efficiently capturing nucleic acids.
  • Figures 9 to 11 show such a surface.
  • Figure 9 shows a top view of such a cell having a wedge shaped inlet and a wedge shaped outlet.
  • Figure 10 shows a magnified view of the microstructured surface of the inner surface of a cell wall.
  • Fig. 11 shows a scanning-electron-microscopic picture of columnar structures of such a surface which provide a high surface-to-volume ratio.
  • Such a surface of analytical cell 11 is a particularly well adapted binding surface for efficiently capturing nucleic acids .
  • An homogeneous spatial distribution of liquid flow through chamber 12 is important in order to achieve a most efficient capture of nucleic acids by the binding surface located within chamber 12.
  • the flow pattern represented in Fig. 6 is more advantageous than the flow pattern represented in Fig. 5.
  • the shape of the inlet and outlet of chamber 12 shown in Fig . 5 yields a higher flow rate through the central part thereof where the flow path is shorter and the flow resistance lower than on paths aside of the central part. Therefore the flow rate is not as homogeneous as desirable.
  • This disadvantage is overcome with the shape of the inlet and outlet of chamber 12 shown in Fig. 6 where the flow path length and the flow resistance is more uniformly distributed and ensures an homogeneous flow of liquid through chamber 12.
  • the hatched area represents the functional binding area.
  • the binding surface within chamber 12 either as part of the chamber walls or as part of a separate body, has chemical properties which are compatible with the chemical substances and reaction conditions necessary for performing a polymerase chain reaction within chamber 12.
  • the binding surface within chamber 12 contains more than 30 weight percent silicon, and more than 35 weight percent oxygen.
  • the binding surface within chamber 12 substantially consists of a metal oxide, e.g. of Ti02, ZnO, Zr02, A1203 or mixtures thereof.
  • the binding surface within chamber 12 substantially consists of a polymer, e.g. of polyamides, polystyrene, polypyrrole, cationic or anionic ion exchange resins.
  • chamber 12 has at least one wall 16 which enables heating and cooling of the contents of chamber 12.
  • the temperature of chamber 12 is thereby modifiable in order to carry out therein a process for amplifying a target nucleic acid sequence which is part of the nucleic acid captured by means of the binding surface.
  • Fig. 12 shows an embodiment of an analytical system according to the invention.
  • This system comprises an analytical device having the features described above and in addition the following components:
  • - thermal instrument means e.g. a thermal cycler, for heating and cooling the contents of chamber 12 of analytical cell 11 in order to carry out an nucleic acid amplification process within chamber 12, and
  • - optical means e.g. a fluorometer 41, for examining the contents of said chamber 12 in order to measure the degree of amplification achieved by a nucleic acid amplifying process performed in said chamber 12 and thereby measuring the amount of an analyte present in the test sample.
  • a thermal cycler used as thermal instrument means has a heat transfer body 51 adapted to contact a surface 18 of wall 16 of chamber 12. This arrangement enables heating and cooling of the contents of said chamber 12 as required for carrying out a process for amplifying a target nucleic acid sequence within chamber 12.
  • Fluorometer 41 is adapted to irradiate the contents of chamber 12 with an excitation light beam 44 provided by a light source 42 and to measure fluorescence light 45 emitted by a tested sample within said chamber 12 by means of a light detection module43.
  • This detection module monitors the fluorescence light energy integrated over the whole area of analytical cell which is accessible to optical examination.
  • a test sample to be analyzed for the presence of a predetermined nucleic acid sequence is acquired.
  • This nucleic acid sequence may originate e. g. from a virus, such as hepatitis B virus (HBV) which is present in a blood sample.
  • Samples to be analyzed may contain nucleic acids from bacteria or particular cells.
  • test sample is necessary in order to perform a method according to the invention on such a sample.
  • the preparation of a test sample is thus a preliminary step.
  • this preliminary step comprises obtaining a test sample containing nucleic acids, e.g. by lysing a biological sample, and mixing it with a highly chaotropic salt in order to enable binding of the sample by a binding surface used in silica based extraction method.
  • the mixture so formed is a test sample on which the method according to the invention is performed.
  • the lysing and mixing steps just mentioned are preferably carried out in container 21 shown in Fig. 1.
  • the following method description focuses on the essential steps of a method according to the invention as performed on a suitable test sample which is obtained by the above-mentioned lysing and mixing steps and which contains nucleic acids.
  • An example of a method for determining the presence or the amount of an analyte in a test sample is carried out by means of an analytical cell 11, an analytical device comprising such a cell, and an analytical system, which are described above with reference to Figures 1 to 12, and comprises the following essential steps :
  • nucleic acids contained in the test sample are captured by a binding surface located within chamber 12,
  • a target nucleic acid sequence which is part of the captured nucleic acids is amplified by a suitable chemical reaction
  • steps (b.l) to (b.3) are performed exclusively in chamber 12.
  • not only the presence of the amplified target nucleic acid sequence is detected, but the amount thereof and thereby the amount of a corresponding analyte is measured, e.g. by means of a fluorimetric measurement .
  • nucleic acids are captured exclusively by the binding surface within analytical cell 11.
  • test sample is forced to flow through chamber 12 in order to effect capturing of nucleic acids contained in the test sample by the binding surface within analytical cell 11.
  • the average flow rate is preferably substantially uniform over a substantial part of the binding surface .
  • a target nucleic acid sequence which is part of the captured nucleic acids is amplified within chamber 12 e.g. by means of a polymerase chain reaction.
  • the necessary reagents for performing a polymerase chain reaction are introduced into chamber 12 by corresponding liquid flow through this chamber.
  • chamber 12 is sealed at both ends and is subject to thermal cycling required for the polymerase chain reaction.
  • the step of amplifying the target nucleic acid sequence is performed in the presence of a fluorescently labeled probe or a fluorescent agent that enables a quantitative measurement of the amplified product.
  • the detection step is performed by measuring the fluorescence intensity of said fluorescently labeled probe or fluorescent agent .
  • Results obtained with the above described method are represented in Fig. 8, which shows a diagram showing the variation of the fluorescence intensity measured with the number of thermal cycles. These results are similar to those obtained with conventional instrument means having separate and independent chambers for performing each of the above mentioned essential method steps, i.e. the capturing, amplification and detection step.
  • PCR polymerase chain reaction
  • amplification methods e.g. : self-sustained sequence replication (3SR) and strand-displacement amplification (SDA) , ligase chain reactio (LCR) , nucleic acid sequence-based amplification (NASB) , QB replicase amplification (QBR) , ligase activated transcription (LAT) , repair chain reaction (RCR) and cycling probe reaction (CRC) .
  • 3SR self-sustained sequence replication
  • SDA strand-displacement amplification
  • LCR ligase chain reactio
  • NNB nucleic acid sequence-based amplification
  • QBR QB replicase amplification
  • LAT repair chain reaction
  • RCR repair chain reaction
  • CRC cycling probe reaction
  • detection of amplified nucleic acid for clinical use relies largely on hybridization of the amplified product and a detection probe labeled with a fluorescent agent .
  • a preferred detection procedure used within the scope of the invention uses a so called TaqMan chemistry technique which is described in the following patent specifications: EP0512334 (Bl) , EP0640828 (Bl) , US6171785, US5994056, US5314809, EP0640828 (Bl) .
  • the detection principle of this technique is based on a hybridization probe having a fluorescence label attached to one end and a fluorescence quencher on the other end of the DNA probe. During the amplification process the quencher and the fluorescence label become separated due to the exonuclease activity of the polymerase and the induced fluorescence increase relates quantitatively to the amount of specific products formed during the amplification.
  • This technique allows highly specific real time monitoring of the amplified target.
  • a detailed example of processing of a sample with a method according to the invention comprises the following steps:
  • a primary probe (le ⁇ copies HBV) was mixed with 600 microliter of a lysing buffer (Guanidine isothiocyanate, GuSCN 4.2 M, pH 7.5) and lysed during 10 minutes. After that 750 microliter of Ethanol (EtOH) were added to the mixture and pumped through analytical cell 11 at a flow rate of 0.5 milliliter per second. Then the cell was flush out with air flow, rinsed with 300 microliter of a washing buffer (70% EtOH, pH 7.5) at a flow rate of 300 microliter per second, and flush out again with air. After this chamber 12 of analytical cell 11 was filled with a ready- to-use PCR reagent mix, the inlet and outlet ports of analytical cell 11 were sealed, and cell 11 was subject to a conventional PCR thermal cycling program for HBV.
  • a lysing buffer Guanidine isothiocyanate, GuSCN 4.2 M, pH 7.5
  • composition of the PCR mix used is e.g. 50 mM Tricine pH 8.3;100mM KOAc pH 7.5 ; 3.0 mM Mn(OAc 2 ); Dimethyl sulfoxide (DMSO) 5%; Glycerin 5%; NTP ' s each 300 uM, Primer HBV1, HBV2 each 150 nM, HBV Probe 100 nM, Polymerase 40 U/reaction.
  • DMSO Dimethyl sulfoxide
  • the conventional PCR thermal cycling program for HBV includes 120s at 95°C, followed by 60 cycles of 20s at 95°C followed by 40s at 59°C.
  • the ramps for heating and cooling have a slope of 1.2°C/s.
  • Flow through chamber 12 is accompanied of a pressure loss which depends of the flow resistance presented by the cell and the viscosity of the liquid flowing through the chamber.
  • a pressure loss of 100 millibar was measured for a liquid viscosity of 1 mPas
  • an average pressure loss of 200 millibar was measured for a liquid viscosity of 4 mPas, with a flow rate of 0.5 milliliter per minute.

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

Abstract

Cette invention concerne un procédé et un dispositif servant à déceler la présence ou à déterminer la quantité d'un analyte dans un échantillon pour essai. Ce procédé consiste : (a) à introduire l'échantillon pour essai dans une chambre (12) ; et (b) à exécuter les étapes suivantes dans ladite chambre (12), lesquelles étapes consistent : (b.1) à capturer les acides nucléiques contenus dans l'échantillon pour essai à l'aide d'une surface de liaison (15) placée à l'intérieur de cette chambre, ladite surface de liaison présentant une forte affinité de liaison pour la capture des acides nucléiques ; (b.2) à amplifier une séquence d'acides nucléiques cible appartenant aux acides nucléiques capturés ; et (b.3) à détecter la présence de la séquence d'acides nucléiques cible amplifiée. Ce dispositif comprend une chambre (12) pourvue d'un orifice d'admission (13) pouvant être hermétiquement fermé et servant à introduire dans la chambre (12) un liquide contenant l'échantillon pour essai, et d'un orifice d'évacuation (14) pouvant être hermétiquement fermé et servant à évacuer le liquide de cette chambre (12). Cette chambre (12) renferme une surface de liaison conçue pour capturer les acides nucléiques contenus dans l'échantillon pour essai lorsque le liquide contenant cet échantillon s'écoule dans la chambre (12). Elle comprend également au moins une paroi qui permet de chauffer et de refroidir les contenus de cette chambre (12), les températures de cette chambre (12) pouvant ainsi varier afin qu'on puisse mettre en oeuvre un procédé d'amplification d'une séquence d'acides nucléiques cible appartenant aux acides nucléiques capturés. Une partie de la chambre (12) comprend une zone permettant d'examiner les contenus de la chambre au moyen d'un dispositif optique servant à déceler la présence d'une séquence d'acides nucléiques cible amplifiée dans cette chambre (12).
PCT/EP2003/005972 2002-06-12 2003-06-04 Procede et dispositif de detection de la presence d'un analyte dans un echantillon pour essai WO2003106031A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP02077317A EP1371419A1 (fr) 2002-06-12 2002-06-12 Procédé et dispositif pour détecter la présence d'un analyte dans un échantillon
EP02077317.2 2002-06-12

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WO2003106031A1 true WO2003106031A1 (fr) 2003-12-24

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US9212977B2 (en) 2005-01-18 2015-12-15 Biocept, Inc. Cell separation using microchannel having patterned posts
US9409166B2 (en) * 2007-12-10 2016-08-09 The Trustees Of The University Of Pennsylvania Integrated PCR reactor for cell lysis, nucleic acid isolation and purification, and nucleic acid amplication related applications
US9414777B2 (en) 2004-07-13 2016-08-16 Dexcom, Inc. Transcutaneous analyte sensor
JP2017106929A (ja) * 2009-07-24 2017-06-15 アコーニ バイオシステムズ インコーポレイテッド フローセルデバイス
US9986942B2 (en) 2004-07-13 2018-06-05 Dexcom, Inc. Analyte sensor
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US10610137B2 (en) 2005-03-10 2020-04-07 Dexcom, Inc. System and methods for processing analyte sensor data for sensor calibration
US10813577B2 (en) 2005-06-21 2020-10-27 Dexcom, Inc. Analyte sensor

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US8158410B2 (en) * 2005-01-18 2012-04-17 Biocept, Inc. Recovery of rare cells using a microchannel apparatus with patterned posts
US7695956B2 (en) 2006-01-12 2010-04-13 Biocept, Inc. Device for cell separation and analysis and method of using
EP1878495A1 (fr) * 2006-07-14 2008-01-16 Roche Diagnostics GmbH Dispositif analytique pour traiter thermiquement un fluide et/ou pour en surveiller une propriété
WO2008006618A1 (fr) * 2006-07-14 2008-01-17 Roche Diagnostics Gmbh Dispositif analytique pour traiter thermiquement un fluide et/ou surveiller une propriété de fluide
EP1878496A1 (fr) * 2006-07-14 2008-01-16 Roche Diagnostics GmbH Appareil pour l'analyse de l'acide nucleique
WO2008012550A2 (fr) 2006-07-28 2008-01-31 Diagnostics For The Real World, Ltd. Dispositif, système et procédé destinés au traitement d'un échantillon
WO2009024773A1 (fr) 2007-08-17 2009-02-26 Diagnostics For The Real World, Ltd Dispositif, système et procédé pour traiter un échantillon
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