EP1159069A1 - Pumpgerät zum transfer mindestens eines fluids in einem wegwerf-behälter - Google Patents

Pumpgerät zum transfer mindestens eines fluids in einem wegwerf-behälter

Info

Publication number
EP1159069A1
EP1159069A1 EP00910910A EP00910910A EP1159069A1 EP 1159069 A1 EP1159069 A1 EP 1159069A1 EP 00910910 A EP00910910 A EP 00910910A EP 00910910 A EP00910910 A EP 00910910A EP 1159069 A1 EP1159069 A1 EP 1159069A1
Authority
EP
European Patent Office
Prior art keywords
compartment
compartments
transfer
arrival
liquid
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Withdrawn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
EP00910910A
Other languages
English (en)
French (fr)
Inventor
Bruno Colin
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.)
Biomerieux SA
Original Assignee
Biomerieux SA
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 Biomerieux SA filed Critical Biomerieux SA
Publication of EP1159069A1 publication Critical patent/EP1159069A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027Containers 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
    • B01L3/50273Containers 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 characterised by the means or forces applied to move the fluids
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0809Geometry, shape and general structure rectangular shaped
    • B01L2300/0816Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00Additional constructional details
    • B01L2300/08Geometry, shape and general structure
    • B01L2300/0861Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0864Configuration of multiple channels and/or chambers in a single devices comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/04Moving fluids with specific forces or mechanical means
    • B01L2400/0475Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0481Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure squeezing of channels or chambers
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01LCHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00Moving or stopping fluids
    • B01L2400/06Valves, specific forms thereof
    • B01L2400/0633Valves, specific forms thereof with moving parts

Definitions

  • the present invention relates to a pumping device to allow the transfer of a fluid sample into a sealed consumable, the transfer taking place between at least a first compartment, called the departure, and at least a second compartment, called the arrival, via a narrowing such as a canal.
  • the invention also relates to a method for implementing such a device.
  • Document WO-A-97/27324 uses substantially the same configuration since the transfer of liquids in a sealed compartment is carried out by the external action exerted on a flexible wall of a compartment.
  • the objective is also identical because it allows amplifications of nucleic acids using PCR technology, without contamination, the device being sealed. It comprises a pumping device to allow the transfer of a fluidic sample into a sealed consumable, the transfer taking place between at least a first compartment, said to start, and at least a second compartment, said to arrive, via a narrowing such as a canal.
  • the position of the departure and arrival compartments is not important according to the invention of the applicants, only the position of the points of intersection is essential, more precisely: "the point of intersection between a departure compartment and the narrowing is positioned in the lower part, and preferably at the lowest level, of this departure compartment, and the point of intersection between an arrival compartment and said narrowing is positioned in the upper part, and preferably at highest level, of this arrival compartment. It is these two characteristics which allow the complete transfer of the liquid contained in the departure compartment to the arrival compartment while playing on gravity. This is not the case with the document cited above since for the supply chambers which contain and which contain either the structural elements for carrying out the amplification or the polymerase enzyme, this point of intersection is not in the lowest position.
  • the map of this international application is not in a vertical position but horizontal, since it is specified an upper surface and a lower surface connected to each other by an edge.
  • This document of the state of the art provides that a compartment can be deformed, so that each deformation activates the transfer of all or part of the sample, this deformation is not due to the deformation of the film only but it is due to the presence of an added cushion on the film, which does not facilitate the manufacture of such a system.
  • the present invention proposes to solve this problem by advantageously positioning, on the one hand, the departure compartment or compartments relative to the arrival compartment or compartments, and on the other hand, the compartments with respect to the transfer channels.
  • the present invention relates to a pumping device to allow the transfer of a fluidic sample into a sealed consumable, the transfer being effected between at least a first compartment, called the start, and at least a second compartment, said of arrival, via a narrowing such as a channel, characterized in that the point of intersection between a starting compartment and the narrowing is positioned in the lower part, and preferably at the lowest level, of this starting compartment , that the point of intersection between an arrival compartment and said narrowing is positioned in the upper part, and preferably at the highest level, of this arrival compartment, and that at least one of the departure compartments and / or arrival can be deformed, so that each deformation activates the transfer of the sample.
  • a narrowing such as a channel
  • This narrowing can be constituted, in addition to a channel, by a pipe of more or less large dimension, whether in length, in diameter or in section, it can also be a simple bottleneck.
  • the only necessity lies in the fact that at the point of intersection of this narrowing with the departure or arrival compartment, the section of the narrowing is less than the section of the compartment.
  • the point of intersection between a departure compartment and the narrowing is in contact with the sample, and the point of intersection between an arrival compartment and said narrowing is not in contact with said sample.
  • the departure compartment or compartments are placed vertically, substantially above the arrival compartment or compartments. The term substantially should be interpreted as meaning that the two compartments are not positioned one above the other with respect to the vertical. The concepts of verticality and horizontality are always given in the present application in relation to the force of gravity.
  • the point of intersection between a departure compartment and the narrowing is in the upper position relative to the point of intersection between an arrival compartment and said narrowing.
  • This upper position must be interpreted as being in a first horizontal plane, containing the first point of intersection mentioned above, in a higher position relative to a second horizontal plane, containing the first point of intersection mentioned above.
  • each compartment which can be deformed comprises at least one partition which can be deformed, such as a flexible film glued on at least one side of the consumable.
  • the consumable is a card that works in an inclined or vertical position, preferably in a vertical position.
  • inclined position is measured in relation to the angle made by the card with a horizontal plane. This angle must be greater than 10 °, advantageously greater than 45 ° and preferably 90 ° which corresponds to the vertical position.
  • each narrowing is traversed longitudinally, in whole or in part, by at least one tab which facilitates the drainage of the fluid sample.
  • At least one of the compartments is associated with a buffer volume.
  • the present invention also relates to a pumping method, which consists in deforming at least once a device as described above.
  • this process consists in deforming at least one of the starting compartments to create a higher than normal pressure in the gas volume of this compartment, in order to transfer all or part of the liquid contained by said liquid in at least one compartment of arrival.
  • this method consists in deforming at least one of the inlet compartments to create an overpressure in the gas volume of this compartment, in order to transfer all or part of the gas contained by said liquid in at least one starting compartment.
  • the process combines the two previous techniques. It alternately consists of:
  • the method in the case where a buffer volume is associated with at least one of the departure and / or arrival compartments, the method consists in simultaneously deforming the departure and / or arrival compartment and the buffer volume.
  • the flexible film when the flexible film is pressed to compress the volume of one of the compartments, it is also possible to simultaneously compress the buffer volume which is associated with it.
  • each container is covered with a flexible film, which can be the same, provided that said film sandwiches said card. It is of course possible to combine this fourth embodiment with the other embodiments.
  • a striker movable between two positions exerting a series of successive pressures on the compartment.
  • This striker can be mobile for example by the action of an electric or pneumatic motor.
  • This striker can be made of any material such as metal, plastic, provided that its mechanical strength allows it to exert pressure or successive pressures on the compartment which can be deformed.
  • the use of such a device relates to a consumable for the analysis of one or more different liquid samples, in which it is sought to identify one or more analytes according to all the simple or complex analysis processes involving one or more different reagents depending on the chemical, physical or biological nature of the analyte (s) sought.
  • the technical principles defined below are not limited to a particular analyte, the only condition being that the analyte is distributed in the sample to be analyzed in suspension or in solution.
  • the analysis process implemented can be carried out, in homogeneous or heterogeneous or mixed form.
  • ligand any biological species such as, for example, an antigen, an antigen fragment, a peptide, antibody, antibody fragment, hapten, nucleic acid, nucleic acid fragment, hormone, vitamin.
  • An example of the application of analytical techniques concerns immunoassays, whatever their format, by direct analysis or by competition.
  • Another example of application relates to the detection and / or quantification of nucleic acids comprising all the operations necessary for this detection and / or this quantification from any sample containing the target nucleic acids.
  • sealed consumable is particularly important in the case where an enzymatic amplification reaction is carried out in the consumable, since the contamination problems linked to these reactions can be avoided by the use of a sealed consumable and that it is particularly advantageous to have a simple system for moving liquids as described in the present invention.
  • the notion of sealed consumable must be understood as a sealed consumable during certain phases of the process and in particular during the phase of displacement of fluids by pumping. In fact, it is necessary to introduce a sample containing one or more ligands to be analyzed into a consumable in order to carry out the analysis of said ligands.
  • the consumable must therefore be opened. Similarly, it is not necessary for the entire consumable to be sealed in order to carry out the invention.
  • the fluid part concerned by the pumping device can be isolated for example by a valve system while another part is open towards the outside to bring reagents to a predetermined place of the consumable for subsequent reactions.
  • FIG. 1 represents a view in partial longitudinal section of a device for pumping a sealed consumable, according to the present invention, in the simplest possible embodiment, since there is a departure compartment and a compartment d 'arrival.
  • Figure 2 shows a partial cross-sectional view of the compartment of a pumping device according to Figure 1
  • FIG. 3 represents a partial longitudinal section view of a device for pumping a sealed consumable, according to the present invention, in a more complex embodiment, since there is a departure compartment and three arrival compartments .
  • FIG. 4 represents a view in longitudinal section of a sealed consumable, which comprises a pumping device according to the present invention, in a complex embodiment substantially identical to FIG. 2, but comprising three important differences. First of all, there is a departure compartment and five arrival compartments. Then, the arrival compartments have a particular configuration. And the layout of the departure compartment compared to the arrival compartments is different from the previous figures.
  • FIG. 5 represents a partial longitudinal section view of a device for pumping a sealed consumable, according to the present invention, in a more complex embodiment, since there are three departure compartments and three arrival compartments .
  • Figure 6 shows a partial cross-sectional view identical to Figure 2, but in which the flexible film of the pumping device is subjected to an external force which induces pumping.
  • the present invention relates to a pumping device 1, well represented in FIGS. 1 to 5, which is particularly well suited to the transfer of fluids, whether in liquid or gaseous form, inside a sealed consumable 3.
  • Figures 1 to 3 and 5 are views in longitudinal section along different embodiments of the pumping device
  • Figure 1 shows a simple embodiment of the present invention. It is characterized by the presence of a first said departure compartment 4 located in the upper part and a second said arrival compartment 5 located in the lower position.
  • the two compartments 4 and 5 are connected to each other by a channel 6, the shape of which is adapted.
  • the volume of this channel 6 must be of a volume adapted to the size of the compartments and the positioning of the compartments between them. Details will be provided later.
  • channel 6 the location of channel 6 relative to the compartment meets certain characteristics.
  • this insertion point is located at the bottom of the start compartment 4 while it is located at the top of the arrival compartment 5.
  • a fluid sample 2 is present in the pumping device 1.
  • This sample 2 is in fact liquid.
  • the point of intersection between the channel 6 and the starting compartment 4 is therefore in contact with the sample 2 which, under the action of gravity, is found at the bottom of said compartment 4.
  • the point of intersection of channel 6 with compartment 5 is in contact with the air contained in said compartment 5, the liquid 2 being present throughout the pipe 6.
  • the fluid sample 2 is therefore present only at the bottom of compartment 5.
  • FIG. 1 there is therefore an alternation between the gas constituted by air and the sample constituted by a liquid 2.
  • the sealed consumable 3 containing the pumping device 1 is partially represented in FIG. 2, which represents a sectional view along AA of FIG. 1. It is noted that it is essentially a card comprising cavities, said cavities being delimited on one side by the material constituting the consumable 3, for example plastic, and on the other side by a very thin partition 7 which is endowed with flexibility, it may be a polyethylene film or any other material which can be deformed, such as silicone, latex, a polyimide.
  • the nature of the flexible film can also vary depending on the nature of the analysis card and the fluids tested, in particular for compatibility reasons.
  • a TPX (polymethylethylpentene) or BOPP (bi-oriented polypropylene) polymer film makes it possible to carry out biological tests.
  • the fixing of these films can be carried out by gluing (coating of glue, such as for example silicone glues on the film) or by welding.
  • glue such as for example silicone glues on the film
  • An example of adhesive BOPP is provided by the company
  • BioMérieux Inc (St Louis, MO, USA) under the reference 022004-2184.
  • the analysis card is obtained by machining a technical plastic material such as impact polystyrene reference R540E from the company GOODFELLOW, compatible with the treated liquids.
  • the card could be obtained by precision molding, but all other manufacturing methods, and in particular those used in semiconductor techniques such as those described in patent application WO-A-97/02357 , can be used for the manufacture of the analysis card.
  • Another condition for operating the device is to have a drop in the liquid when this liquid is at the intersection of the channels
  • this system is also found at the level of the inlet compartment 5 only and that the pumping is then carried out by actuating a film 7 present only at the level of this inlet compartment 5.
  • the volume of departure from the inlet compartment, once compressed, is reduced and the air is transferred via channel 6 to the departure compartment 4.
  • the volume of exhaust air of the arrival compartment 5 is large enough to reach the departure compartment 4.
  • this volume of air displaced must be greater than the volume of channel 6.
  • FIG. 3 another embodiment is shown comprising a single starting compartment 4 and three arrival compartments 5.
  • the channel 6 has a particular structure, since there is a primary channel 8 for each compartment 4 or 5 and an intermediate channel 9 making it possible to make the connection between all of the primary channels 8.
  • the shape of the different channels such as channels 6, 8, 9, 10 and 11, must have a shape adapted to the various embodiments, and in particular when a distribution function exists in the system with a multitude of inlet compartments, the person skilled in the art will choose a shape for each channel allowing distribution balanced including by playing on the pressure drops.
  • Elbows, narrowing are, for example, means to modulate this pressure drop and not to favor the preferential filling of an inlet compartment.
  • a person skilled in the art can play on these forms of channels to promote filling.
  • the size and nature of the channel will also be chosen to allow the liquid-gas exchange necessary for the operation of the pumping device.
  • the section of the narrowing channel 6 will be adapted as a function of the viscosity of the liquid to allow the air bubble to rise. It is particularly advantageous to minimize the volume of the narrowing channel 6 relative to the volume of liquid to be transferred to avoid dead volumes, especially in the case where the starting compartment is not located above, compared to the concept of gravity. , of the arrival compartment (s).
  • valves also makes it possible to control the filling of the inlet compartments possibly in combination with the shape of the channels.
  • These valves can have different functions such as for example a distribution function used to direct the fluid in a particular direction, to replace the distribution channel 11 in FIG. 4 or a closing / opening function to isolate the card or a compartment such as on valve 15 in Figure 4.
  • Figure 5 is also a particular embodiment in which there are three starting compartments 4 and three arrival compartments 5.
  • the channel or narrowing 6 is then of a slightly more complex structure since there is for each compartment 4 or 5 a primary channel 8, as already shown in FIG. 3, but also an intermediate channel 9 for each group of departure 4 or arrival compartments 5.
  • the two intermediate channels 9 are positioned parallel l to each other. Between these two channels 9, there are present two so-called secondary channels 10.
  • the number of these secondary channels 10 is absolutely not imposed. There may be one or more.
  • the two channels 10 are in fact arranged to allow a better distribution of the liquid samples 2, which are transferred from the departure compartments 4 to the arrival compartments 5. This type of reaction also requires a good quantification of the volumes transferred.
  • the present invention allows such a controlled distribution. This function will be explained later.
  • the present invention should therefore be understood as being capable of reversing the role of the gas and the liquid since each compartment, at one time of the pumping process, is partially filled with one and the other constituent.
  • the starting compartment may be full of liquid and the arrival compartment full of gas.
  • said inlet compartment already contains at least one substance, which may be liquid and / or solid. It may for example be a coating containing at least one reagent intended to react with the transferred sample 2.
  • FIG. 4 represents an embodiment substantially closer to the industrial models which can be manufactured. It is a system substantially identical to FIG. 3 since there is a single departure compartment 4 and five arrival compartments 5.
  • Figures 3 and 4 there are also substantial differences between these Figures 3 and 4.
  • the starting compartment 4 is located below the arrival compartments 5.
  • the embodiments of Figures 1 to 3 and 5 are therefore particularly interesting.
  • a second difference lies in the structure of the arrival compartments. These include in the upper position a device 13 for breaking bubbles which has also been the subject of a patent application by the applicant as indicated above.
  • the total volume of liquid transferable by this device can vary from 0.5 to 5000 microliters, advantageously from 2 to 2000 microliters and preferably from 5 to 1000 microliters. In the case of a large volume, such as for example greater than 500 microliters, it is preferable to choose an embodiment by successive pressure for transferring fractions of liquid with a volume of between 5 and 100 microliters ( ⁇ l).
  • the volume of the starting compartment varies in the same proportions or can be significantly greater than the total volume to be transferred.
  • the chamber 4 has a volume of between 2 and 5 ml for a transfer of liquid of between 250 and 500 ⁇ l.
  • the total volume for the fluid part represented by the channel 8 connecting the starting compartment 4 and the inlet compartment 5 is 120 ⁇ l broken down between 20 ⁇ l for the part of the channel 8 connecting the compartment 5 and the distribution channel 11 and 5 times 20 ⁇ l for the part of the channel 8 connecting the distribution channel 11 and the compartments 5
  • the channel 8 in this example is of semicircular section with a diameter of 0.5 mm.
  • the transfer from another consumable 3 as well as the valves 15 have already been the subject of a patent application filed by the plaintiff on September 8, 1998, under the filing number FR98 / 11383, and entitled: "Device allowing reactions, transfer system between devices and method of implementing such a system ”. The content of the description of this patent application is considered to be contained in the present invention.
  • inlets 12, outlets 16 and valves 15 are not described in relation to FIGS. 1 to 3, 5 and 6, it is of course obvious that these pumping devices 1 are equipped with them, even if this is not not shown in the figures, in order to allow them to function properly.

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)
  • Automatic Analysis And Handling Materials Therefor (AREA)
  • Feeding, Discharge, Calcimining, Fusing, And Gas-Generation Devices (AREA)
  • Sampling And Sample Adjustment (AREA)
EP00910910A 1999-03-09 2000-03-09 Pumpgerät zum transfer mindestens eines fluids in einem wegwerf-behälter Withdrawn EP1159069A1 (de)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
FR9903031 1999-03-09
FR9903031A FR2790681B1 (fr) 1999-03-09 1999-03-09 Dispositif de pompage permettant de transferer au moins un fluide dans un consommable
PCT/FR2000/000580 WO2000053320A1 (fr) 1999-03-09 2000-03-09 Dispositif de pompage permettant de transferer au moins un fluide dans un consommable

Publications (1)

Publication Number Publication Date
EP1159069A1 true EP1159069A1 (de) 2001-12-05

Family

ID=9543082

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00910910A Withdrawn EP1159069A1 (de) 1999-03-09 2000-03-09 Pumpgerät zum transfer mindestens eines fluids in einem wegwerf-behälter

Country Status (6)

Country Link
EP (1) EP1159069A1 (de)
JP (1) JP2003517368A (de)
AU (1) AU3295200A (de)
CA (1) CA2362739A1 (de)
FR (1) FR2790681B1 (de)
WO (1) WO2000053320A1 (de)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7704728B2 (en) * 2001-07-18 2010-04-27 The University Of Michigan Microfluidic gravity pump with constant flow rate
JP4208820B2 (ja) 2003-11-28 2009-01-14 株式会社東芝 核酸検出カセット
US8173078B2 (en) * 2004-04-28 2012-05-08 Industrial Technology Research Institute Gravity-driven micropump
CN102042954B (zh) * 2010-10-22 2012-06-27 重庆大学 一种包衣薄膜与基层之间粘附力的静压鼓泡测量方法
CN102072875B (zh) * 2010-10-26 2012-11-28 重庆大学 一种静压鼓泡试验中包衣薄膜弹性应变能的几何测量法
CN102297836B (zh) * 2011-05-18 2012-11-14 重庆大学 一种包衣薄膜与基层之间界面粘附强度的直接测量方法

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EP0192794B1 (de) * 1985-02-27 1988-06-22 Horiba, Ltd. Flüssigkeitsbehälter für Analysegeräte
US4761381A (en) * 1985-09-18 1988-08-02 Miles Inc. Volume metering capillary gap device for applying a liquid sample onto a reactive surface
WO1994026414A1 (en) * 1993-05-17 1994-11-24 Syntex (U.S.A.) Inc. Reaction container for specific binding assays and method for its use
IL115327A (en) * 1994-10-07 2000-08-13 Bayer Ag Diaphragm pump
US5863502A (en) * 1996-01-24 1999-01-26 Sarnoff Corporation Parallel reaction cassette and associated devices
FR2749663B1 (fr) * 1996-06-07 1998-07-31 Bio Merieux Carte d'analyse a usage unique comprenant un conduit d'ecoul ement de liquides
US5811296A (en) * 1996-12-20 1998-09-22 Johnson & Johnson Clinical Diagnostics, Inc. Blocked compartments in a PCR reaction vessel
JP3582316B2 (ja) * 1997-08-20 2004-10-27 株式会社日立製作所 化学分析装置

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Title
See references of WO0053320A1 *

Also Published As

Publication number Publication date
FR2790681B1 (fr) 2001-05-11
FR2790681A1 (fr) 2000-09-15
JP2003517368A (ja) 2003-05-27
CA2362739A1 (fr) 2000-09-14
WO2000053320A1 (fr) 2000-09-14
AU3295200A (en) 2000-09-28

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