EP1371419A1 - Procédé et dispositif pour détecter la présence d'un analyte dans un échantillon - Google Patents

Procédé et dispositif pour détecter la présence d'un analyte dans un échantillon Download PDF

Info

Publication number
EP1371419A1
EP1371419A1 EP02077317A EP02077317A EP1371419A1 EP 1371419 A1 EP1371419 A1 EP 1371419A1 EP 02077317 A EP02077317 A EP 02077317A EP 02077317 A EP02077317 A EP 02077317A EP 1371419 A1 EP1371419 A1 EP 1371419A1
Authority
EP
European Patent Office
Prior art keywords
chamber
test sample
nucleic acid
nucleic acids
analytical device
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP02077317A
Other languages
German (de)
English (en)
Inventor
Martin Kopp
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
F Hoffmann La Roche AG
Roche Diagnostics GmbH
Original Assignee
F Hoffmann La Roche AG
Roche Diagnostics GmbH
Boehringer Mannheim 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, Boehringer Mannheim GmbH filed Critical F Hoffmann La Roche AG
Priority to EP02077317A priority Critical patent/EP1371419A1/fr
Priority to PCT/EP2003/005972 priority patent/WO2003106031A1/fr
Publication of EP1371419A1 publication Critical patent/EP1371419A1/fr
Withdrawn legal-status Critical Current

Links

Images

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 6.
  • a system according to the invention is defined by claim 18.
  • analyte refers to a sequence of nucleic acid, e. g. DNA or RNA, whose presence in a test sample is to be detected.
  • 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.
  • 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 16 has an outer surface 18 which during the intended use of the cell is in thermal contact with heating or cooling means of 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.
  • a part of chamber 12 has a zone 17 which allows examination of the chamber contents by optical means for detecting the presence of an amplified target nucleic acid sequence contained in chamber 12.
  • 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: Element Atom % Weight % O 57.26 43.15 Na 8.41 9.11 Al 3.36 4.27 Si 27.14 35.91 K 1.84 3.40 Ca 0.88 1.66 Ti 1.11 2.50
  • SiO x and Al 2 O 3 are particularly important components of the fleece with regard to its capacity to bind nucleic acids.
  • a fleece consisting of pure SiO 2 shows an acceptable capacity to bind nucleic acids.
  • 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 TiO2, ZnO, ZrO2, Al2O3 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:
  • 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:
  • 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.
  • 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
  • SDA self-sustained sequence replication
  • LCR ligase chain reaction
  • NNB nucleic acid sequence-based amplification
  • QBR QB replicase amplification
  • LAT ligase activated transcription
  • 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 (B1), EP0640828 (B1), US6171785, US5994056, US5314809, EP0640828 (B1).
  • 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.
  • 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: 150 microliter of a primary probe (1e6 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
  • 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.

Landscapes

  • 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)
EP02077317A 2002-06-12 2002-06-12 Procédé et dispositif pour détecter la présence d'un analyte dans un échantillon Withdrawn EP1371419A1 (fr)

Priority Applications (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
PCT/EP2003/005972 WO2003106031A1 (fr) 2002-06-12 2003-06-04 Procede et dispositif de detection de la presence d'un analyte dans un echantillon pour essai

Applications Claiming Priority (1)

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

Publications (1)

Publication Number Publication Date
EP1371419A1 true EP1371419A1 (fr) 2003-12-17

Family

ID=29558391

Family Applications (1)

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

Country Status (2)

Country Link
EP (1) EP1371419A1 (fr)
WO (1) WO2003106031A1 (fr)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005084545A1 (fr) * 2004-03-06 2005-09-15 Roche Diagnostics Gmbh Dispositif de prelevement de fluides corporels
US20060160243A1 (en) 2005-01-18 2006-07-20 Biocept, Inc. Recovery of rare cells using a microchannel apparatus with patterned posts
WO2006078470A2 (fr) 2005-01-18 2006-07-27 Biocept, Inc. Separation de cellules utilisant un microcanal comportant des tiges disposees selon un motif particulier
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
EP1878496A1 (fr) * 2006-07-14 2008-01-16 Roche Diagnostics GmbH Appareil pour l'analyse de l'acide nucleique
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
US7695956B2 (en) 2006-01-12 2010-04-13 Biocept, Inc. Device for cell separation and analysis and method of using
US7819822B2 (en) 2004-03-06 2010-10-26 Roche Diagnostics Operations, Inc. Body fluid sampling device
EP2714884A1 (fr) * 2011-05-27 2014-04-09 The University Of British Columbia Appareil microfluidique de piégeage et d'essai cellulaires pour analyse à haut rendement
US9212977B2 (en) 2005-01-18 2015-12-15 Biocept, Inc. Cell separation using microchannel having patterned posts
US9707556B2 (en) 2007-08-17 2017-07-18 Diagnostics For The Real World, Ltd. Device, system and method for processing a sample
US9839909B2 (en) 2006-07-28 2017-12-12 Diagnostics For The Real World, Ltd. Device, system and method for processing a sample

Families Citing this family (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7920906B2 (en) 2005-03-10 2011-04-05 Dexcom, Inc. System and methods for processing analyte sensor data for sensor calibration
US9247900B2 (en) 2004-07-13 2016-02-02 Dexcom, Inc. Analyte sensor
WO2006127694A2 (fr) 2004-07-13 2006-11-30 Dexcom, Inc. Detecteur d'analyte
US8989833B2 (en) 2004-07-13 2015-03-24 Dexcom, Inc. Transcutaneous analyte sensor
EP1666150B1 (fr) 2004-11-20 2015-01-07 Roche Diagnostics GmbH Préparation d'acide nucléique
WO2009002580A2 (fr) * 2007-04-02 2008-12-31 Boston Medical Center Corporation Procédé de lyse bactérienne
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
WO2011011062A2 (fr) * 2009-07-24 2011-01-27 Akonni Biosystems Dispositif de cuve à circulation

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993022053A1 (fr) * 1992-05-01 1993-11-11 Trustees Of The University Of Pennsylvania Structures de detection micro-usinees
EP0693560A2 (fr) * 1994-07-19 1996-01-24 Becton, Dickinson and Company Procédé et appareil pour amplification automatique d'acides nucleiques essai sur acide nucleique et immunoessai
WO2000060362A1 (fr) * 1999-04-08 2000-10-12 Chung Chang Young Appareil de preparation et d'analyse rapides d'acides nucleiques
US20020018998A1 (en) * 2000-02-11 2002-02-14 Michael Fritz System for simple nucleic acid analysis

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2018274T5 (es) * 1986-03-11 1996-12-16 Plant Genetic Systems Nv Celulas vegetales resistentes a los inhibidores de glutamina sintetasa, preparadas por ingenieria genetica.
EP1175431A4 (fr) * 1999-04-30 2002-09-11 Whatman Inc Substrat contenant un detergent anionique de purification d'acide nucleique

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993022053A1 (fr) * 1992-05-01 1993-11-11 Trustees Of The University Of Pennsylvania Structures de detection micro-usinees
EP0693560A2 (fr) * 1994-07-19 1996-01-24 Becton, Dickinson and Company Procédé et appareil pour amplification automatique d'acides nucleiques essai sur acide nucleique et immunoessai
WO2000060362A1 (fr) * 1999-04-08 2000-10-12 Chung Chang Young Appareil de preparation et d'analyse rapides d'acides nucleiques
US20020018998A1 (en) * 2000-02-11 2002-02-14 Michael Fritz System for simple nucleic acid analysis

Cited By (33)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8000762B2 (en) * 2004-03-06 2011-08-16 Roche Diagnostics Operations, Inc. Body fluid sampling device
EP2727531A3 (fr) * 2004-03-06 2018-06-27 Roche Diabetes Care GmbH Dispositif d'échantillonnage de fluide corporel
WO2005084545A1 (fr) * 2004-03-06 2005-09-15 Roche Diagnostics Gmbh Dispositif de prelevement de fluides corporels
US9022952B2 (en) 2004-03-06 2015-05-05 Roche Diagnostics Operations, Inc. Body fluid sampling device
EP1722670A2 (fr) * 2004-03-06 2006-11-22 F.Hoffmann-La Roche Ag Dispositif d'echantillonnage de liquide corporel
US8814808B2 (en) 2004-03-06 2014-08-26 Roche Diagnostics Operations, Inc. Body fluid sampling device
EP2705792A1 (fr) * 2004-03-06 2014-03-12 F. Hoffmann-La Roche AG Dispositif d'échantillonnage de fluide corporel
EP1722670B1 (fr) * 2004-03-06 2013-09-25 F.Hoffmann-La Roche Ag Dispositif d'echantillonnage de liquide corporel
US8369918B2 (en) 2004-03-06 2013-02-05 Roche Diagnostics Operations, Inc. Body fluid sampling device
US8162854B2 (en) 2004-03-06 2012-04-24 Roche Diagnostics Operations, Inc. Body fluid sampling device
US7819822B2 (en) 2004-03-06 2010-10-26 Roche Diagnostics Operations, Inc. Body fluid sampling device
CN103382434B (zh) * 2005-01-18 2016-05-25 生物概念股份有限公司 利用含排列成图案的立柱的微通道分离细胞
WO2006078470A3 (fr) * 2005-01-18 2006-09-14 Biocept Inc Separation de cellules utilisant un microcanal comportant des tiges disposees selon un motif particulier
US10369568B2 (en) 2005-01-18 2019-08-06 Biocept, Inc. Cell separation using microchannel having patterned posts
US20060160243A1 (en) 2005-01-18 2006-07-20 Biocept, Inc. Recovery of rare cells using a microchannel apparatus with patterned posts
CN101102847B (zh) * 2005-01-18 2013-07-17 生物概念股份有限公司 利用含排列成图案的立柱的微通道分离细胞
WO2006078470A2 (fr) 2005-01-18 2006-07-27 Biocept, Inc. Separation de cellules utilisant un microcanal comportant des tiges disposees selon un motif particulier
CN103382434A (zh) * 2005-01-18 2013-11-06 生物概念股份有限公司 利用含排列成图案的立柱的微通道分离细胞
US9212977B2 (en) 2005-01-18 2015-12-15 Biocept, Inc. Cell separation using microchannel having patterned posts
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
WO2008006500A1 (fr) * 2006-07-14 2008-01-17 Roche Diagnostics Gmbh Appareil d'analyse d'acide nucléique
EP1878496A1 (fr) * 2006-07-14 2008-01-16 Roche Diagnostics GmbH Appareil pour l'analyse de l'acide nucleique
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
US9839909B2 (en) 2006-07-28 2017-12-12 Diagnostics For The Real World, Ltd. Device, system and method for processing a sample
US10315195B2 (en) 2006-07-28 2019-06-11 Diagnostics For The Real World, Ltd. Device, system and method processing a sample
US9707556B2 (en) 2007-08-17 2017-07-18 Diagnostics For The Real World, Ltd. Device, system and method for processing a sample
US10661271B2 (en) 2007-08-17 2020-05-26 Diagnostics For The Real World, Ltd. Device, system and method for processing a sample
US9902990B2 (en) 2011-05-27 2018-02-27 The University Of British Columbia Microfluidic cell trap and assay apparatus for high-throughput analysis
EP2714884A1 (fr) * 2011-05-27 2014-04-09 The University Of British Columbia Appareil microfluidique de piégeage et d'essai cellulaires pour analyse à haut rendement
EP2714884A4 (fr) * 2011-05-27 2015-04-22 Univ British Columbia Appareil microfluidique de piégeage et d'essai cellulaires pour analyse à haut rendement
US10508299B2 (en) 2011-05-27 2019-12-17 The University Of British Columbia Microfluidic cell trap and assay apparatus for high-throughput analysis

Also Published As

Publication number Publication date
WO2003106031A1 (fr) 2003-12-24

Similar Documents

Publication Publication Date Title
EP1371419A1 (fr) Procédé et dispositif pour détecter la présence d'un analyte dans un échantillon
JP6838127B2 (ja) 統合された移送モジュールを有する試験カートリッジ
Chen et al. Total nucleic acid analysis integrated on microfluidic devices
AU2009201529B2 (en) Apparatus For Polynucleotide Detection and Quantitation
US20090061450A1 (en) System and method for diagnosis of infectious diseases
CN1143917A (zh) 用于测定和加工分析物的中尺度样品制备设备和系统
WO2014093714A1 (fr) Procédés et compositions pour l'utilisation d'huiles pour l'analyse et la détection de molécules
US8092999B2 (en) Biological sample reaction chip and biological sample reaction method
WO2014000037A1 (fr) Kit d'amplification et de détection d'acide nucléique
Li et al. Recent advancements in nucleic acid detection with microfluidic chip for molecular diagnostics
AU2007225103A1 (en) System and method for diagnosis of infectious diseases
CN106854674B (zh) 一种基于毛细管微阵列的核酸高通量快速检测方法
Chen et al. An integrated microfluidic loop-mediated isothermal amplification platform for koi herpesvirus detection
JP2018520706A (ja) 試料採取からｎｇｓライブラリ調製までの自動化
van Kooten et al. Purely electrical SARS-CoV-2 sensing based on single-molecule counting
WO2016044673A1 (fr) Amplification d'acide nucléique multi-étapes hybride
İnce et al. Micro-polymerase chain reaction for point-of-care detection and beyond: A review microfluidics and nanofluidics
WO2009149115A1 (fr) Cartouche pour effectuer des essais biochimiques
WO2000077253A1 (fr) Appareil et procede d'examen des genes
Hoang et al. Fluidic system with movable layers for all-in-one assay of cell-free DNA in blood
McMillan et al. Application of advanced microfluidics and rapid PCR to analysis of microbial targets
WO2023034995A1 (fr) Procédé de perçage d'un joint d'étanchéité pour test d'échantillon
Juang et al. Oil Immersed Lossless Total Analysis System (OIL-TAS): Integrated RNA Extraction and Detection for SARS-CoV-2 Testing
Malewski et al. RT-PCR technique and its applications. State-of the-art
Gärtner et al. A microfluidic toolbox approach to CBRNE sensing

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE TR

AX Request for extension of the european patent

Extension state: AL LT LV MK RO SI

AKX Designation fees paid
REG Reference to a national code

Ref country code: DE

Ref legal event code: 8566

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20040618