EP1740296B1 - Procede pour produire une solution, dispositif correspondant et utilisations du procede et du dispositif - Google Patents
Procede pour produire une solution, dispositif correspondant et utilisations du procede et du dispositif Download PDFInfo
- Publication number
- EP1740296B1 EP1740296B1 EP05738068A EP05738068A EP1740296B1 EP 1740296 B1 EP1740296 B1 EP 1740296B1 EP 05738068 A EP05738068 A EP 05738068A EP 05738068 A EP05738068 A EP 05738068A EP 1740296 B1 EP1740296 B1 EP 1740296B1
- Authority
- EP
- European Patent Office
- Prior art keywords
- solvent
- solid matter
- cavity
- soluble
- pcr
- 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.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title claims description 23
- 239000007787 solid Substances 0.000 claims abstract description 60
- 239000002904 solvent Substances 0.000 claims abstract description 43
- 239000003153 chemical reaction reagent Substances 0.000 claims description 16
- 238000004458 analytical method Methods 0.000 claims description 10
- 239000012188 paraffin wax Substances 0.000 claims description 8
- 239000011324 bead Substances 0.000 claims description 7
- 239000012807 PCR reagent Substances 0.000 claims description 6
- 238000003752 polymerase chain reaction Methods 0.000 claims 5
- 239000002671 adjuvant Substances 0.000 claims 1
- 239000000872 buffer Substances 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 239000002773 nucleotide Substances 0.000 claims 1
- 125000003729 nucleotide group Chemical group 0.000 claims 1
- 239000013615 primer Substances 0.000 claims 1
- 239000002987 primer (paints) Substances 0.000 claims 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 1
- 239000000243 solution Substances 0.000 description 11
- 239000007788 liquid Substances 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 3
- 238000002844 melting Methods 0.000 description 3
- 230000008018 melting Effects 0.000 description 3
- 230000001681 protective effect Effects 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 239000010410 layer Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000009993 protective function Effects 0.000 description 2
- 239000010409 thin film Substances 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- 238000012742 biochemical analysis Methods 0.000 description 1
- 239000012482 calibration solution Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000000155 melt Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000011241 protective layer Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000005382 thermal cycling Methods 0.000 description 1
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
- B01F35/71—Feed mechanisms
- B01F35/713—Feed mechanisms comprising breaking packages or parts thereof, e.g. piercing or opening sealing elements between compartments or cartridges
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F33/00—Other mixers; Mixing plants; Combinations of mixers
- B01F33/30—Micromixers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F35/00—Accessories for mixers; Auxiliary operations or auxiliary devices; Parts or details of general application
- B01F35/71—Feed mechanisms
- B01F35/713—Feed mechanisms comprising breaking packages or parts thereof, e.g. piercing or opening sealing elements between compartments or cartridges
- B01F35/7135—Opening the seal between the compartments by application of heat
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
- B01F21/00—Dissolving
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/25—Chemistry: analytical and immunological testing including sample preparation
Definitions
- the invention relates to a method and associated arrangement for producing a solution of at least one solid in a solvent.
- the invention relates to applications of the arrangement or the method, in particular in the chemical investigations.
- solids are dissolved in a solvent.
- Such solids are, for example, reagents which are stored as a dry mixture with negligible vapor pressure and form a storage-stable substance at room temperature. Only when needed, they are resolved for the intended use in the analysis.
- Such a device is used for example in the WO 02/0072262 A1 described.
- an analysis device designed as an easy-to-handle chip card, are stored in the dry reagents as solids and dissolved in a solvent as needed.
- the dry reagents present in the chip card can already be stored in predosed and pre-portioned form for the analysis.
- a solution with a precisely defined amount of reagent is produced in conjunction with a solvent from a reservoir provided from the pre-portioned reagents.
- a disposable detector array for real-time fluid analysis in which a liquid sample is automatically analyzed by a disposable sensor and the associated readings are output. It is essential that the analysis reagent is kept ready as a liquid.
- the preparation of a solution of at least one solid in a solvent can be targeted. It is advantageous that this process can take place in a closed unit and that in this unit the solid is stored.
- the first solid is a PCR reagent.
- a second solid may be isolated DNA.
- DNA is known to be so-called magnetic beads annealed which are suspended in the solvent. They are enriched in a channel or a cavity in which a PCR is to take place.
- the invention is therefore intended in particular for carrying out the PCR. But other analysis applications that provide solids and solve them as needed are also possible.
- an examination unit is designated 1.
- This unit is a so-called cartridge body, which may be part of a portable device.
- a cavity 2 is present, in which certain reactions can take place.
- fluidic channels from which a first channel 3 leads to the cavity 2 and a second channel 3 'leads away from the cavity 2. It can be valves or The same may be present with which the Fluidigkanäle 3 and 3 'are lockable at a suitable location.
- a first solid 5 is stored.
- the solid 5 is a reagent or a mixture of reagents, wherein the mixture forms a storage-stable substance at room temperature.
- the solid 5 forms a layer on the bottom of the cavity 2.
- the surface of the solid 5 is protected by a medium 6 forming a thin film on the solid layer 5.
- the medium 6 is not soluble in the solvent in which the solid 5 is to be dissolved.
- Paraffin may advantageously be used as the medium 6, which itself is not soluble in an aqueous solvent. Other insoluble in the solvent media are possible. Instead of a thin film which predominantly covers the first solid 2-dimensionally, a 3-dimensional coating of e.g. spherical solid particles immobilized in the cavity can be used.
- FIG. 2 It is shown that a solvent 7 flows through the fluidic channel 3 and fills the cavity or flows through it.
- the protected by the medium 6 solid 5 remains unaffected by the solvent.
- the protective medium 6 is paraffin in particular, it can be dissolved by heating. This is in FIG. 3 shown. After the melting of the paraffin, the medium 6 no longer has a protective function. It forms, for example, individual beads, which are indicated as 61 and 61 '. On the other hand, the solid has migrated into solution or suspension 8, so that the dissolved reagent is located in cavity 2 for further uses.
- FIG. 4 otherwise the FIG. 2 corresponds to, such a solvent 17 is introduced into the cavity 2, which contains a second solid 19.
- the first solid 5 is in the FIG. 4 corresponding Figure 1/2 is protected by the medium 6.
- the second solid 19 is either dissolved in the solvent or it is present as a suspension.
- a liquid substance can also be dissolved or present as an emulsion in the solvent.
- FIG. 5 After heating and melting of the paraffin, the medium 6 no longer has a protective function.
- FIG. 5 will turn accordingly FIG. 3 Beads 61 and 61 'of the medium formed.
- a solution or suspension 18 is now formed with the first solid 5 and the second solid 19, which is available for further uses. If the so-called second solid is a liquid, an emulsion will accordingly be formed.
- First and second solids can be chosen so that they can react with each other after removal of the protective medium and, if necessary, their properties, e.g. Solubility can change.
- first solid is a PCR reagent and the second solid is DNA
- PCR reactions can occur in the well.
- suitable means for thermal cycling are necessary.
- Paraffin is particularly suitable here as a protective medium, since in the PCR a first heating process is required anyway and thereby melts the paraffin protective layer.
- hot-start PCR, which prevents the PCR enzyme polymerase from becoming unspecific and therefore disadvantageously effective even below a certain temperature (paraffin melting temperature).
- the DNA as a second solid is usually bound in the prior art to so-called magnetic beads and is thus present as a suspension.
- the magnetic bead-bound DNA can be collected by magnetic means and selectively brought into the cavity 2.
- the above-described method and the associated device are particularly suitable for carrying out a PCR. But also for other applications in biomedical technology, especially if at certain times, especially immediately before an analysis to be carried out, quantitative solutions of individual dry reagents are to be formed, the invention is applicable.
- Examples of the preparation of defined reagent solutions include an enzyme-label solution, an enzyme-substrate solution or general calibration solutions.
Landscapes
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Claims (20)
- Procédé de préparation d'une solution ( 8, 18 ) d'au moins une substance ( 5, 19 ) solide dans un solvant ( 7, 17 ), le solvant ( 7, 17 ) étant envoyé d'un réservoir à une cavité ( 2 ) par des micro-canaux ( 3, 3' ) ayant les mesures suivantes :- on emmagasine dans la cavité ( 2 ) la substance ( 5, 19 ) solide soluble dans le solvant ( 7, 17 ) et on la recouvre ou on l'enrobe d'un milieu ( 6 ) insoluble dans le solvant ( 7 ), de manière à empêcher une dissolution de la substance ( 5, 19 ) solide soluble dans le solvant ( 7, 17 ),- on fait passer dans la cavité ( 2 ) un agent de lavage,- on envoie le solvant ( 7, 17 ) dans la cavité ( 2 ),- on traite le milieu insoluble dans le solvant ( 7, 17 ) de façon à permettre un contact entre le solvant ( 7 ) et la substance ( 5, 19 ) solide insoluble et ainsi une dissolution de la substance ( 5, 19 ) solide dans le solvant ( 7 ) et à former la solution.
- Procédé suivant la revendication 1, dans lequel il y a deux substances ( 5, 19 ) solides et la première substance ( 5 ) solide est mise en réserve à l'avance dans la cavité ( 2 ) et la deuxième substance ( 19 ) est contenue dans le solvant ( 17 ), ayant les autres mesures suivantes :- on recouvre ou on enrobe la première substance ( 5 ) solide soluble dans le solvant ( 17 ) dans la cavité ( 2 ) d'un milieu ( 6 ) insoluble dans le solvant ( 17 ) de manière à empêcher une dissolution de la substance solide soluble dans le solvant ( 17 ),- on envoie dans la cavité ( 2 ) le solvant ( 17 ) contenant la deuxième substance ( 19 ) solide.
- Procédé suivant la revendication 1 ou suivant la revendication 2, caractérisé en ce que l'on effectue une intégration complète de tous les stades du procédé dans une unité qui peut être utilisée une seule fois et dans laquelle au moins une substance ( 5, 19 ) solide est mise en réserve à l'avance dans la cavité ( 2 ).
- Procédé suivant la revendication 1 ou la revendication 2, caractérisé en ce que l'on ferme la cavité ( 2 ) par des vannes.
- Procédé suivant les revendications 1 à 3, caractérisé en ce que l'on utilise de l'eau comme solvant ( 7 ).
- Procédé suivant la revendication 2, caractérisé en ce que l'on met la deuxième substance ( 19 ) solide en suspension dans le solvant ( 17 ).
- Procédé suivant l'une des revendications précédentes, caractérisé en ce que la première substance ( 5 ) solide est un réactif PCR.
- Procédé suivant la revendication 7, caractérisé en ce que le réactif ( 5 ) PCR est constitué de polymérase, de nucléotides, d'amorces, de tampons ainsi que d'adjuvants.
- Procédé suivant la revendication 8, caractérisé en ce que le réactif ( 5 ) PCR forme un film sur la paroi de la cavité ( 2 ).
- Procédé suivant la revendication 2 et la revendication 6, caractérisé en ce que la deuxième substance ( 19 ) solide est de l'ADN à amplifier dissous ou en suspension.
- Procédé suivant la revendication 10, caractérisé en ce que des ADN ( 19 ) à amplifier sont fixés à des perles magnétiques.
- Procédé suivant la revendication 11, caractérisé en ce qu'il est prévu des moyens de rassemblement de perles magnétiques dans la cavité ( 2 ) PCR.
- Procédé suivant l'une des revendications précédentes, caractérisé en ce que le milieu ( 6 ) qui n'est pas soluble dans le solvant est de la paraffine.
- Dispositif pour la mise en oeuvre du procédé suivant la revendication 1 ou l'une des revendications 2 à 4, comprenant une unité qui est réalisée en produit à jeter, au moins un micro-canal ( 3, 3' ) ou une microcavité ( 2 ) étant prévu pour la réception d'au moins une substance ( 5, 19 ) solide soluble dans un solvant, au moins la première substance ( 5, 19 ) solide se trouvant sur une paroi de la microcavité ( 2 ) et étant recouverte d'une mince couche d'un milieu insoluble dans le solvant.
- Dispositif suivant la revendication 14, caractérisé en ce que la cavité est une cavité ( 2 ) PCR appropriée à une thermocyclisation.
- Dispositif suivant la revendication 14, caractérisé en ce qu'il y a des moyens de thermocyclisation.
- Dispositif suivant la revendication 15, caractérisé en ce que la cavité ( 2 ) PCR est équipée d'une entrée ( 3 ) et d'une sortie ( 3' ) .
- Dispositif suivant la revendication 15, caractérisé en ce que l'entrée ( 3 ) et la sortie ( 3' ) sont équipées de vannes.
- Utilisation du dispositif suivant la revendication 14, dans une PCR ( Polymerase Chain Reaction ) pour des études biochimiques.
- Utilisation du dispositif suivant la revendication 14, dans une dissolution quantitative de réactif à sec dans le solvant pour préparer un réactif pour un dispositif d'analyse.
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE102004021821A DE102004021821B3 (de) | 2004-04-30 | 2004-04-30 | Verfahren zur Herstellung einer Lösung, zugehörige Anordnung sowie Anwendungen dieser Anordnung |
PCT/EP2005/051870 WO2005105284A1 (fr) | 2004-04-30 | 2005-04-26 | Procede pour produire une solution, dispositif correspondant et utilisations du procede et du dispositif |
Publications (2)
Publication Number | Publication Date |
---|---|
EP1740296A1 EP1740296A1 (fr) | 2007-01-10 |
EP1740296B1 true EP1740296B1 (fr) | 2008-10-08 |
Family
ID=34966122
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP05738068A Expired - Fee Related EP1740296B1 (fr) | 2004-04-30 | 2005-04-26 | Procede pour produire une solution, dispositif correspondant et utilisations du procede et du dispositif |
Country Status (5)
Country | Link |
---|---|
US (1) | US8518343B2 (fr) |
EP (1) | EP1740296B1 (fr) |
CN (1) | CN1946473B (fr) |
DE (2) | DE102004021821B3 (fr) |
WO (1) | WO2005105284A1 (fr) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE102004050575B3 (de) * | 2004-10-15 | 2006-01-05 | Siemens Ag | Verfahren zur kombinierten Isolierung von Magnet-Beads aus einer flüssigen Probe und anschließender Thermozyklisierung für die PCR und zugehörige Anordnung |
JP5786295B2 (ja) * | 2010-06-22 | 2015-09-30 | ソニー株式会社 | 核酸等温増幅反応用マイクロチップ及びその製造方法並びに核酸等温増幅方法 |
DE102018200520A1 (de) | 2018-01-15 | 2019-07-18 | Robert Bosch Gmbh | Verfahren zum Bereitstellen einer Lösung der Substanz in einer mikrofluidischen Vorrichtung |
US11666914B2 (en) * | 2018-05-09 | 2023-06-06 | Tecan Trading Ag | Cartridge, electrowetting sample processing system and bead manipulation method |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US1572323A (en) * | 1922-12-16 | 1926-02-09 | Claude A Smith | Medicinal and anesthetic package |
US1603877A (en) * | 1925-05-18 | 1926-10-19 | Smith Arthur Ervin | Medicinal package |
US5096669A (en) | 1988-09-15 | 1992-03-17 | I-Stat Corporation | Disposable sensing device for real time fluid analysis |
EP1023593A1 (fr) * | 1998-08-19 | 2000-08-02 | JENOPTIK Aktiengesellschaft | Dispositif permettant de transporter de tres petites quantites de fluide et procede de fabrication associe |
DE10111457B4 (de) * | 2001-03-09 | 2006-12-14 | Siemens Ag | Diagnoseeinrichtung |
US7318912B2 (en) * | 2001-06-07 | 2008-01-15 | Nanostream, Inc. | Microfluidic systems and methods for combining discrete fluid volumes |
US7332348B2 (en) * | 2003-02-28 | 2008-02-19 | Applera Corporation | Sample substrate having a divided sample chamber and method of loading thereof |
-
2004
- 2004-04-30 DE DE102004021821A patent/DE102004021821B3/de not_active Expired - Fee Related
-
2005
- 2005-04-26 CN CN2005800126865A patent/CN1946473B/zh not_active Expired - Fee Related
- 2005-04-26 US US11/587,572 patent/US8518343B2/en active Active
- 2005-04-26 WO PCT/EP2005/051870 patent/WO2005105284A1/fr not_active Application Discontinuation
- 2005-04-26 EP EP05738068A patent/EP1740296B1/fr not_active Expired - Fee Related
- 2005-04-26 DE DE502005005620T patent/DE502005005620D1/de active Active
Also Published As
Publication number | Publication date |
---|---|
DE502005005620D1 (de) | 2008-11-20 |
CN1946473B (zh) | 2010-05-26 |
CN1946473A (zh) | 2007-04-11 |
US20070212708A1 (en) | 2007-09-13 |
WO2005105284A1 (fr) | 2005-11-10 |
US8518343B2 (en) | 2013-08-27 |
EP1740296A1 (fr) | 2007-01-10 |
DE102004021821B3 (de) | 2005-12-08 |
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