WO1991018110A1 - Procede et systeme pour effectuer des reactions biochimiques - Google Patents

Procede et systeme pour effectuer des reactions biochimiques Download PDF

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Publication number
WO1991018110A1
WO1991018110A1 PCT/SE1991/000343 SE9100343W WO9118110A1 WO 1991018110 A1 WO1991018110 A1 WO 1991018110A1 SE 9100343 W SE9100343 W SE 9100343W WO 9118110 A1 WO9118110 A1 WO 9118110A1
Authority
WO
WIPO (PCT)
Prior art keywords
capillary
reagent
reaction vessel
bore
reagents
Prior art date
Application number
PCT/SE1991/000343
Other languages
English (en)
Inventor
Mats Malmquist
Original Assignee
Mats Malmquist
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 Mats Malmquist filed Critical Mats Malmquist
Priority to RU9192016293A priority Critical patent/RU2082754C1/ru
Priority to DE69124236T priority patent/DE69124236T2/de
Priority to EP91910180A priority patent/EP0530283B1/fr
Priority to AU79715/91A priority patent/AU660615B2/en
Priority to CA002082933A priority patent/CA2082933C/fr
Publication of WO1991018110A1 publication Critical patent/WO1991018110A1/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
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above

Definitions

  • the present invention relates to a method to perform biochemical reactions and a combination of a capillary and a reaction vessel for use in said method.
  • the invention is applicable for all small volume biochemical reactions in which the reagents cannot be mixed beforehand. Particularly, the invention is intended for the PCR (Polymerase Chain Reaction)-technique.
  • RNA DNA
  • ELISA Enzyme Linked ImmunoSorbent
  • a person may, however, be HIV-positive without antibodies being present if he/she, for instance, is in the early stages of the disease. In this case the ELISA test gives a negative result and the person concerned then risks unwittingly transmitting the infection to others. Therefore the need for a better, i.e. more sensitive, HIV test is very great.
  • the diagnosis of other viruses also, the culture of which previously took a long time, has been improved with the PCR technique.
  • PCR diagnosis comprises three stages:
  • stage 1) is time-consuming and demanding work, primarily because the reagents cannot be mixed in advance, and thus give rise to many sources of error. It is very important that stage 1) should be carried out with great care and precision because the amplification in stage 2) and the detection result in stage 3) depend absolutely on the reliability of stage 1).
  • the object of the invention was to deminish the contamination risk as well as the time required to prepare small reagent volumes for a specific biochemical reaction in which the reagents cannot be mixed in advance and the preparation is time-consuming.
  • a capillary which contains several reagents separated by intermediate hydrophobic liquid, e.g. paraffines, oils, alkanes.
  • reagent storage as well as sample reaction takes place.
  • the sample is added to the capillary and then the capillary is melted at one end.
  • the mixing of the sample with the reagents is done in that a steel pin is put into the capillary and a magnet is moved in an upward and downward direction along the outside of the capillary.
  • the capillary is centrifugated to obtain the reaction solution and the hydrophobic liquid in two separate phases.
  • the capillary has to be cut at the sealed end and also at the boundary between hydrophobic liquid-reaction solution and thereafter the reaction solution is transferred to a cuvette or the like, for measurement of, for example, UV absorbance.
  • Fig. 1 is a diagrammatic view if a reaction vessel including a reagent capillary containing reagent
  • Fig. 2 is a diagrammatic view of an alternative embodiment of a reaction vessel including an alternative embodiment of a reagent capillary;
  • Fig. 3 is a plan view of the embodiment shown in Fig. 2;
  • Fig. 4 shows the reagent capillary depicted in Fig. 1 on a larger scale
  • Fig. 5 shows the reagent capillary depicted in Fig. 2 on a larger scale.
  • Fig. 1 shows a ready-prepared reaction vessel 1 according to the present invention.
  • a reagent capillary 3 Inside the reaction vessel 1, for instance an Eppendorf tube, is placed a reagent capillary 3.
  • the reagent capillary 3 is provided with different reagents, which can be of any suitable type for a desired reaction.
  • In the bottom of the reaction vessel 1 there may be water or buffer 2 for subsequent dilution of the reagents.
  • Fig. 2 shows an alternative and preferred embodiment of a reaction vessel 1 and a reagent capillary 3.
  • the reaction vessel 1 is provided with a lid 4 having a bore 4a.
  • the bore 4a is covered by a permeable membrane 4b.
  • the bore 4a fitted with a membrane is located centrally in the lid 4 in the shown em ⁇ bodiment but this is not a critical feature. In fact it is possible to provide the lid with several bores to be able to put in more than one reagent capillary as desired.
  • the bore 4a forms a stop collar for the reagent capillary 3 in accordance with Fig. 5, which is described in greater detail below.
  • the reagent capillary 3 depicted in Fig. 4 is designed to be inserted into the reagent vessel 1 shown in Fig. 1.
  • the reagent capillary 3 is provided with different reagents 6-13 for a specific biochemical reaction. The amount of each reagent is calculated and intended only for this specific reaction. If a PCR reaction is to be performed the reagent solutions 6-13 comprise PCR buffer, dCTP, dGTP, dATP, dTTP, two or more oligonucleotides, all of the reagents being calculated for a specific PCR reaction, and termostable DNA polymerase. Between the reagents there is air or an inert fluid. Naturally, the mutual order is optional.
  • a modified reagent capillary is depicted in Fig. 5.
  • This reagent capillary is designed to be inserted into a reaction vessel according to Fig. 2.
  • the reagent capillary differs from the reagent capillary shown in Fig. 4 in that there is an annular locking groove 5 on the lower part of the capillary intended to be snapped into the bore 4a.
  • a protective cover 15 is fitted over the upper end of the capillary.
  • the reaction vessel according to Fig. 2 and the reagent capillary according to Fig. 5 are stored separately until use.
  • the lower end of the capillary 3 is pushed through the permeable membrane 4b in the lid 4 of the reaction vessel 1, whereupon the locking groove 5 engages with the stop collar formed by the bore 4a.
  • the protec ⁇ tive cover 15 protects the contents of the reagent capillary 3 from contamination during the process of insertion and pushing into the reaction vessel 1.
  • the reagent solutions in the reagent capillary 3 have thawed, they are then centrifuged down and mixed with one another and, where applicable, with the diluent 2 at the bottom of the vessel 1. After centrifuging, the lid 4 may be opened without having to remove the capillary 3 from the lid.
  • the advantage of this is that material can readily be added to or extracted from the reaction vessel if desired.
  • reagent capillaries After producing the reagent capillaries, i.e. by aspirating the different reagents with air or inert fluid in between, either manually or automatically, they may be packed separately or placed in a reaction vessel in kits for performing a specific biochemical reaction. Of course, this packaging takes place under sterile conditions.
  • An alternative method of producing the capillaries would be to aspirate the reagents into capillaries with air or inert fluid between the reagents, freeze the capillaries, cut the capillaries in the air sections, and to place the desired capillary pieces in one common outer capillary having an inner diameter correspon ⁇ ding to the outer diameter of the capillary pieces. This would allow combining of the reagents in any desired way.
  • the reagent capillaries with or without the reaction vessels are stored in the frozen state until use.
  • the reagent capillary is thawed and the contents thereof are centrifuged down in the reaction vessel, being mixed with each other and with diluent if any.
  • all the reagents, including heat stable DNA polymerase are now in the reaction vessel and the only further addition needed before the amplification is of the sample, e.g. blood.
  • the sample is added by using a dosing system described in applicants pending Swedish patent application SE 91 0726-0. Briefly, the sample is drawn up into a capillary, of the same type as reaction capillary 3, by the capillary effects. Thereaf ⁇ ter the sample capillary is inserted in an unoccupied bore 4a in the lid 4 of the reaction vessel 1 which thereafter is once again centrifugated.
  • the present inventor suggest using material for the reaction vessels that does not or only slightly absorb UV light. If ethidium bromide is added after the reaction it would then be possible to detect whether or not DNA has been amplified by viewing the vessels under UV light with the naked eye. Preferably the ethidium bromide addition is made in the same manner as the above described sample addition.
  • reagent capillaries 3 have been prepared for a specific sample volume and a specific biochemical reaction. Other reactions require different volumes and number of reagents.
  • kits i.e. complete sets containing reagents for a specific reaction.
  • kits usually consist of Eppendorf tubes containing different reagents suitable for about 100 standard reactions.
  • For each reaction a certain volume is mixed from each tube.
  • one kit can contain, e.g. 500 capillaries, each ready to use for the reaction it is designed for.
  • kits based in the reagent capillaries desribed in the present application is that the user does not have to pipette the reagent and is able, instead, to select the appropriate reagent capillary for the relevant reaction with fingers of tweezers.
  • the simplification of the work is obvious, above all in regard to the handling of radioactive reagents, as there is no risk of contaminating pipettes, less risk of radioactive waste and shorter periods of exposure for the staff.
  • reaction capillaries in PCR technology there is the saving in time and the benefits of worker protection in many biotech- nological sectors.

Landscapes

  • 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)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Devices For Use In Laboratory Experiments (AREA)

Abstract

Cette invention concerne un procédé permettant d'effectuer des réactions biochimiques ainsi que l'association d'un tube capillaire (3) et d'un réacteur (1) utlisée dans ce procédé. Les tubes capillaires (3) de réactifs renferment des réactifs gelés (6-13) séparés les uns des autres par de l'air ou un liquide inerte. Lors de l'utilisation, on place les tubes capillaires de réactifs (3) dans le réacteur (1) afin qu'ils dégèlent puis le contenu est centrifugé vers le fond du réacteur (1). Cette invention peut être utilisée pour tous les types de réactions biochimiques courantes et pour tous les types de tests de diagnostic dans lesquels on ne peut mélanger à l'avance les réactifs. Ce procédé est tout particulièrement adapté aux diagnostics selon la technique de l'amplification enzymatique du génome, il est également très utile lorsqu'il s'agit de manipuler des réacitfs radioactifs, par exemple, des nucléotides marqués, au niveau de réactions de séquençage,etc.
PCT/SE1991/000343 1990-05-16 1991-05-15 Procede et systeme pour effectuer des reactions biochimiques WO1991018110A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
RU9192016293A RU2082754C1 (ru) 1990-05-16 1991-05-15 Способ проведения биохимических реакций и устройство для его осуществления
DE69124236T DE69124236T2 (de) 1990-05-16 1991-05-15 Verfahren und vorrichtung zur ausführung biochemischer reaktionen
EP91910180A EP0530283B1 (fr) 1990-05-16 1991-05-15 Procede et systeme pour effectuer des reactions biochimiques
AU79715/91A AU660615B2 (en) 1990-05-16 1991-05-15 Method and means to perform biochemical reactions
CA002082933A CA2082933C (fr) 1990-05-16 1991-05-15 Methode et moyens mis en oeuvre pour obtenir des reactions biochimiques

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE9001772A SE465086B (sv) 1990-05-16 1990-05-16 Reagenskapillaerer, reaktionskaerl, beredningssaett och anvaendning daerav
SE9001772-4 1990-05-16

Publications (1)

Publication Number Publication Date
WO1991018110A1 true WO1991018110A1 (fr) 1991-11-28

Family

ID=20379508

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SE1991/000343 WO1991018110A1 (fr) 1990-05-16 1991-05-15 Procede et systeme pour effectuer des reactions biochimiques

Country Status (9)

Country Link
EP (1) EP0530283B1 (fr)
JP (1) JPH0646936B2 (fr)
AT (1) ATE147789T1 (fr)
AU (1) AU660615B2 (fr)
CA (1) CA2082933C (fr)
DE (1) DE69124236T2 (fr)
RU (1) RU2082754C1 (fr)
SE (1) SE465086B (fr)
WO (1) WO1991018110A1 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993015222A1 (fr) * 1992-01-29 1993-08-05 Institut National De La Sante Et De La Recherche Medicale (Inserm) Procede pour eviter les risques de contamination, notamment applicable aux techniques d'amplification de l'adn ou de l'arn
WO1993016200A1 (fr) * 1992-02-13 1993-08-19 Kosak Kenneth M REACTIFS THERMO-LIBERABLES DESTINES AUX REACTIONS CHIMIQUES $i (IN VITRO)
EP0572057A1 (fr) * 1992-05-11 1993-12-01 Johnson & Johnson Clinical Diagnostics, Inc. Composition et kit de réactifs de PCR et procédés pour amplification et détection avec amplification non-spécifique d'acides nucléiques réduite
WO1995019447A1 (fr) * 1994-01-14 1995-07-20 The Jockey Club Procede d'echantillonnage non invasif pour analyse d'acides nucleiques
WO1997048491A1 (fr) * 1996-06-20 1997-12-24 Hamilton Bonaduz Ag Procede pour effectuer des reactions chimiques, notamment biochimiques, et pointe de pipette avec recipient reactionnel, eventuellement avec element de reception supplementaire pour la pointe de pipette
WO1998054292A1 (fr) * 1997-05-28 1998-12-03 Alphahelix Ab Nouvelle cuve a reaction et ses procedes d'utilisation
ES2153745A1 (es) * 1998-07-31 2001-03-01 Ivia Dispositivo compartimentado y metodo para la captura y doble amplificacion enzimatica de secuencias diana de acidos nucleicos en un solo dispositivo compartimentado cerrado.
US6511814B1 (en) 1999-03-26 2003-01-28 Idexx Laboratories, Inc. Method and device for detecting analytes in fluids
US6551842B1 (en) 1999-03-26 2003-04-22 Idexx Laboratories, Inc. Method and device for detecting analytes in fluids
US6602719B1 (en) 1999-03-26 2003-08-05 Idexx Laboratories, Inc. Method and device for detecting analytes in fluids

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH11509100A (ja) * 1995-07-13 1999-08-17 イムノロジカル アソシエーツ オブ デンバー 核酸抽出、増幅および検出を統合する自己充足装置
EP0843176B1 (fr) * 1995-07-31 2013-06-12 Precision System Science Co., Ltd. Système d' analyse comprenant un recipient et un élément d' aspiration et de distribution de liquide
JP5899624B2 (ja) * 2011-02-18 2016-04-06 セイコーエプソン株式会社 反応容器
US10589191B2 (en) * 2011-06-24 2020-03-17 Shimadzu Corporation Method for separating substance accommodated in vessel

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DD225788A1 (de) * 1984-04-25 1985-08-07 Univ Berlin Humboldt Mikrotest zur durchfuehrung analytischer bestimmungen

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DD225788A1 (de) * 1984-04-25 1985-08-07 Univ Berlin Humboldt Mikrotest zur durchfuehrung analytischer bestimmungen

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993015222A1 (fr) * 1992-01-29 1993-08-05 Institut National De La Sante Et De La Recherche Medicale (Inserm) Procede pour eviter les risques de contamination, notamment applicable aux techniques d'amplification de l'adn ou de l'arn
WO1993016200A1 (fr) * 1992-02-13 1993-08-19 Kosak Kenneth M REACTIFS THERMO-LIBERABLES DESTINES AUX REACTIONS CHIMIQUES $i (IN VITRO)
US5413924A (en) * 1992-02-13 1995-05-09 Kosak; Kenneth M. Preparation of wax beads containing a reagent for release by heating
US5643764A (en) * 1992-02-13 1997-07-01 Kosak; Kenneth M. Reactions using heat-releasable reagents in wax beads
EP0572057A1 (fr) * 1992-05-11 1993-12-01 Johnson & Johnson Clinical Diagnostics, Inc. Composition et kit de réactifs de PCR et procédés pour amplification et détection avec amplification non-spécifique d'acides nucléiques réduite
WO1995019447A1 (fr) * 1994-01-14 1995-07-20 The Jockey Club Procede d'echantillonnage non invasif pour analyse d'acides nucleiques
WO1997048491A1 (fr) * 1996-06-20 1997-12-24 Hamilton Bonaduz Ag Procede pour effectuer des reactions chimiques, notamment biochimiques, et pointe de pipette avec recipient reactionnel, eventuellement avec element de reception supplementaire pour la pointe de pipette
WO1998054292A1 (fr) * 1997-05-28 1998-12-03 Alphahelix Ab Nouvelle cuve a reaction et ses procedes d'utilisation
US6451258B1 (en) 1997-05-28 2002-09-17 Alphahelix Ab Reaction vessel, cassette and system for performing biochemical reactions
ES2153745A1 (es) * 1998-07-31 2001-03-01 Ivia Dispositivo compartimentado y metodo para la captura y doble amplificacion enzimatica de secuencias diana de acidos nucleicos en un solo dispositivo compartimentado cerrado.
US6511814B1 (en) 1999-03-26 2003-01-28 Idexx Laboratories, Inc. Method and device for detecting analytes in fluids
US6551842B1 (en) 1999-03-26 2003-04-22 Idexx Laboratories, Inc. Method and device for detecting analytes in fluids
US6602719B1 (en) 1999-03-26 2003-08-05 Idexx Laboratories, Inc. Method and device for detecting analytes in fluids

Also Published As

Publication number Publication date
SE9001772A (fr) 1991-07-22
DE69124236T2 (de) 1997-08-07
ATE147789T1 (de) 1997-02-15
CA2082933C (fr) 2002-09-17
SE465086B (sv) 1991-07-22
AU7971591A (en) 1991-12-10
AU660615B2 (en) 1995-07-06
JPH04228100A (ja) 1992-08-18
RU2082754C1 (ru) 1997-06-27
EP0530283B1 (fr) 1997-01-15
CA2082933A1 (fr) 1991-11-17
EP0530283A1 (fr) 1993-03-10
DE69124236D1 (de) 1997-02-27
JPH0646936B2 (ja) 1994-06-22
SE9001772D0 (sv) 1990-05-16

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