EP0530283B1 - Procede et systeme pour effectuer des reactions biochimiques - Google Patents

Procede et systeme pour effectuer des reactions biochimiques Download PDF

Info

Publication number
EP0530283B1
EP0530283B1 EP91910180A EP91910180A EP0530283B1 EP 0530283 B1 EP0530283 B1 EP 0530283B1 EP 91910180 A EP91910180 A EP 91910180A EP 91910180 A EP91910180 A EP 91910180A EP 0530283 B1 EP0530283 B1 EP 0530283B1
Authority
EP
European Patent Office
Prior art keywords
capillary
reagent
reaction vessel
bore
reagents
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 - Lifetime
Application number
EP91910180A
Other languages
German (de)
English (en)
Other versions
EP0530283A1 (fr
Inventor
Mats Malmquist
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.)
Individual
Original Assignee
Individual
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 Individual filed Critical Individual
Publication of EP0530283A1 publication Critical patent/EP0530283A1/fr
Application granted granted Critical
Publication of EP0530283B1 publication Critical patent/EP0530283B1/fr
Anticipated expiration legal-status Critical
Expired - Lifetime 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
    • B01L3/508—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 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 embodiment 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 protective 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 corresponding 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. If a PCR reaction is to be performed, 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. Thereafter 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 biotechnological 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)
  • Devices For Use In Laboratory Experiments (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)

Claims (12)

  1. Procédé de mise en oeuvre de réactions biochimiques, dans lequel les réactifs ne peuvent pas être mélangés au préalable, dans lequel un élément capillaire réactif (3) est utilisé, introduit avec des réactifs séparés les uns des autres, caractérisé en ce que l'élément capillaire réactif (3) est inséré dans un premier alésage (4a) ménagé dans le couvercle (4) d'un récipient de réaction (1), en ce que le récipient de réaction (1) ayant un élément capillaire réactif (3) inséré est centrifugé pour amener le contenu de l'élément capillaire réactif (3) au fond du récipient de réaction (1), et en ce que l'échantillon devant être mis en réaction est ajouté au récipient de réaction (1).
  2. Procédé selon la revendication 1, caractérisé en ce qu'un élément capillaire contenant l'échantillon est inséré dans un deuxième alésage (4a) ménagé dans le couvercle (4) du récipient de réaction (1), et en ce qu'une deuxième centrifugation est effectuée.
  3. Procédé selon la revendication 1 ou 2, caractérisé en ce que les solutions réactives (6 à 13) sont réfrigérées et en ce qu'elles sont dégelées avant centrifugation.
  4. Procédé selon la revendication 1, 2 ou 3, caractérisé en ce qu'un couvercle protecteur (15) est monté sur l'éxtrémité supérieure de l'élément capillaire réactif (3), avant que son extrémité inférieure soit insérée dans l'alésage (4a), l'extrémité inférieure étant insérée dans le récipient de réaction (1) jusqu'à ce qu'une gorge de verrouillage (5) située sur l'élément capillaire s'engage contre les bords de l'alésage (4a).
  5. Procédé selon la revendication 4, caractérisé en ce que le réactif capillaire (3) est inséré dans une membrane perméable (4b) recouvrant l'alésage (4a).
  6. Combinaison d'un élément capillaire réactif et d'un récipient de réaction, caractérisée par un élément capillaire réactif (3) comprenant différentes solutions réactives (6 à 13) en des volumes prédéterminés séparés les uns des autres par de l'air ou un fluide inerte, et un récipient de réaction (1) comprenant un couvercle (4) pourvu d'un ou plusieurs alésages (4a), l'élément capillaire réactif (3) étant destiné à être inséré dans l'alésage (4a) ménagé dans le couvercle (4) du récipient de réaction (1) en utilisation.
  7. Combinaison selon la revendication 6, caractérisée en ce que les solutions réactives (6 à 13) sont congelées et en ce qu'elles sont dégelées avant que l'élément capillaire réactif (3) soit inséré dans l'alésage (4a).
  8. Combinaison selon la revendication 6 ou 7, caractérisée en ce que l'élément capillaire réactif (3) est pourvu d'une gorge de verrouillage (5) à son extrémité inférieure et d'un couvercle protecteur (15) à son extrémité supérieure.
  9. Combinaison selon la revendication 6, 7 ou 8, caractérisée en ce que l'alésage (4a) est recouvert d'une membrane perméable (4b).
  10. Combinaison selon une ou plusieurs des revendications 6 à 9, caractérisée en ce que les solutions réactives (6 à 13) comprennent un ou plusieurs acide(s) nucléique(s) et/ou enzyme(s) pour une réaction specifique.
  11. Combinaison selon la revendication 10, caractérisée en ce que les solutions réactives (6 à 13) comprennent un tampon PCR, dCTP, dGTP, dATP, dTTP, deux ou plusieurs oligonucléotides, tous les réactifs étant calculés pour une réaction PCR spécifique, et une polymérase ADN thermostable.
  12. Combinaison selon une ou plusieurs des revendications 5 à 11, caractérisée en ce que le récipient de réaction (1) est constitué d'un matériau qui n'absorbe pas ou absorbe seulement faiblement la lumière UV.
EP91910180A 1990-05-16 1991-05-15 Procede et systeme pour effectuer des reactions biochimiques Expired - Lifetime EP0530283B1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
SE9001772 1990-05-16
SE9001772A SE465086B (sv) 1990-05-16 1990-05-16 Reagenskapillaerer, reaktionskaerl, beredningssaett och anvaendning daerav
PCT/SE1991/000343 WO1991018110A1 (fr) 1990-05-16 1991-05-15 Procede et systeme pour effectuer des reactions biochimiques

Publications (2)

Publication Number Publication Date
EP0530283A1 EP0530283A1 (fr) 1993-03-10
EP0530283B1 true EP0530283B1 (fr) 1997-01-15

Family

ID=20379508

Family Applications (1)

Application Number Title Priority Date Filing Date
EP91910180A Expired - Lifetime EP0530283B1 (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)

Families Citing this family (14)

* 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
US5413924A (en) * 1992-02-13 1995-05-09 Kosak; Kenneth M. Preparation of wax beads containing a reagent for release by heating
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
DE69637047T2 (de) * 1995-07-13 2007-12-27 Applera Corp., Foster City Unabhängiges gerät zur extraktion, amplifikation und nachweis von nukleinsäuren
JP3985872B2 (ja) * 1995-07-31 2007-10-03 プレシジョン・システム・サイエンス株式会社 容器
DE19624687A1 (de) * 1996-06-20 1998-01-29 Hamilton Bonaduz Ag Verfahren zur Durchführung chemischer, insbesondere biochemischer Reaktionen und Pipettierspitze, ggf. mit Reaktionsgefäß
SE9702005D0 (sv) * 1997-05-28 1997-05-28 Alphahelix Ab New reaction vessel and method for its use
ES2153745B1 (es) * 1998-07-31 2001-11-16 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
JP5899624B2 (ja) * 2011-02-18 2016-04-06 セイコーエプソン株式会社 反応容器
WO2012176598A1 (fr) * 2011-06-24 2012-12-27 株式会社島津製作所 Procédé pour séparer une substance reçue dans un récipient

Family Cites Families (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

Also Published As

Publication number Publication date
CA2082933A1 (fr) 1991-11-17
SE465086B (sv) 1991-07-22
ATE147789T1 (de) 1997-02-15
SE9001772A (fr) 1991-07-22
DE69124236D1 (de) 1997-02-27
SE9001772D0 (sv) 1990-05-16
AU660615B2 (en) 1995-07-06
JPH04228100A (ja) 1992-08-18
CA2082933C (fr) 2002-09-17
DE69124236T2 (de) 1997-08-07
RU2082754C1 (ru) 1997-06-27
AU7971591A (en) 1991-12-10
WO1991018110A1 (fr) 1991-11-28
JPH0646936B2 (ja) 1994-06-22
EP0530283A1 (fr) 1993-03-10

Similar Documents

Publication Publication Date Title
US5846489A (en) System for opening closures of vessels and for the contamination-free operation of reaction sequences
EP0530283B1 (fr) Procede et systeme pour effectuer des reactions biochimiques
Belak et al. Experiences on the application of the polymerase chain reaction in a diagnostic laboratory
US20060275915A1 (en) Method and apparatus for mixing sample and reagent in a suspension fluid
US20090061450A1 (en) System and method for diagnosis of infectious diseases
US20060258010A1 (en) Apparatus and method for handling fluids for analysis
CA2173861A1 (fr) Tube de reaction et methode d'utilisation de celui-ci minimisant les contaminations
EP1108006A1 (fr) Dispositif assurant a lui seul l'extraction, l'amplification et la detection d'acides nucleiques
US6428962B1 (en) Nucleic acid collection barrier method and apparatus
KR20150018774A (ko) 통합형 전달 모듈을 구비한 테스트 카트리지
EP1371419A1 (fr) Procédé et dispositif pour détecter la présence d'un analyte dans un échantillon
US5273907A (en) Method and means for perform biochemical reactions
US4960219A (en) Snap cap
US5821436A (en) Method for the repeated transfer of liquids
EP1628770B1 (fr) Eprouvette pour recueillir et preparer des echantillons pour amplification par reaction en chaine de la polymerase
US6200781B1 (en) Apparatus, system and method for automated execution and analysis of biological and chemical reactions
NZ517565A (en) Direct aspiration-reaction and injection device and methods of use
EP4043887B1 (fr) Décapsuleur permettant d'enlever un capuchon d'un récipient d'échantillons de laboratoire
Truchaud et al. Increasing the biosafety of analytical systems in the clinical laboratory
Juang et al. Oil Immersed Lossless Total Analysis System (OIL-TAS): Integrated RNA Extraction and Detection for SARS-CoV-2 Testing
KR20240056605A (ko) 분자진단 시스템의 샘플 처리 및 분석 방법
AU777999B2 (en) Direct aspiration-reaction and injection device and methods of use
CN115261183A (zh) 一种即用型基因检测包
Manual AmpliSens® Mycoplasma hominis-EPh

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

17P Request for examination filed

Effective date: 19921116

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LI LU NL SE

17Q First examination report despatched

Effective date: 19950217

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LI LU NL SE

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IT

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT;WARNING: LAPSES OF ITALIAN PATENTS WITH EFFECTIVE DATE BEFORE 2007 MAY HAVE OCCURRED AT ANY TIME BEFORE 2007. THE CORRECT EFFECTIVE DATE MAY BE DIFFERENT FROM THE ONE RECORDED.

Effective date: 19970115

Ref country code: AT

Effective date: 19970115

Ref country code: LI

Effective date: 19970115

Ref country code: DK

Effective date: 19970115

Ref country code: ES

Free format text: THE PATENT HAS BEEN ANNULLED BY A DECISION OF A NATIONAL AUTHORITY

Effective date: 19970115

Ref country code: CH

Effective date: 19970115

Ref country code: GR

Free format text: LAPSE BECAUSE OF FAILURE TO SUBMIT A TRANSLATION OF THE DESCRIPTION OR TO PAY THE FEE WITHIN THE PRESCRIBED TIME-LIMIT

Effective date: 19970115

REF Corresponds to:

Ref document number: 147789

Country of ref document: AT

Date of ref document: 19970215

Kind code of ref document: T

REG Reference to a national code

Ref country code: CH

Ref legal event code: EP

REF Corresponds to:

Ref document number: 69124236

Country of ref document: DE

Date of ref document: 19970227

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: SE

Effective date: 19970415

ET Fr: translation filed
PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: LU

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 19970531

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

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

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

26N No opposition filed
REG Reference to a national code

Ref country code: GB

Ref legal event code: IF02

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 20030430

Year of fee payment: 13

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: BE

Payment date: 20030507

Year of fee payment: 13

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: GB

Payment date: 20030509

Year of fee payment: 13

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 20030528

Year of fee payment: 13

Ref country code: DE

Payment date: 20030528

Year of fee payment: 13

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: GB

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20040515

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: BE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20040531

BERE Be: lapsed

Owner name: *MALMQUIST MATS

Effective date: 20040531

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NL

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20041201

Ref country code: DE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20041201

GBPC Gb: european patent ceased through non-payment of renewal fee

Effective date: 20040515

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: FR

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20050131

NLV4 Nl: lapsed or anulled due to non-payment of the annual fee

Effective date: 20041201

REG Reference to a national code

Ref country code: FR

Ref legal event code: ST