EP2277052A2 - Device for detecting at least one element contained in a blood solution - Google Patents
Device for detecting at least one element contained in a blood solutionInfo
- Publication number
- EP2277052A2 EP2277052A2 EP09757728A EP09757728A EP2277052A2 EP 2277052 A2 EP2277052 A2 EP 2277052A2 EP 09757728 A EP09757728 A EP 09757728A EP 09757728 A EP09757728 A EP 09757728A EP 2277052 A2 EP2277052 A2 EP 2277052A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- detection
- solution
- zone
- blood
- receiving
- 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- Withdrawn
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/80—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood groups or blood types or red blood cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
Definitions
- the present invention relates to a device for detecting at least one element contained in a solution at least partly constituted by at least one component of the blood.
- the invention relates to the field of manufacturing devices designed to detect at least one element contained in a blood solution, in particular consisting of a patient's blood sample or serum contained in this blood.
- devices are known in the form of a card, in particular cardboard or plastic, having a test zone comprising a test means operating on the principle of a conventional agglutination reaction.
- Such a device has many disadvantages. Indeed, in addition to the fact that such a device is impractical to implement and unhygienic (the blood + reactive agglutination mixture is likely to flow), the use of this device to perform a test of Blood group requires compliance with a particular experimental protocol which, when not respected, leads to results that may prove to be wrong. This is particularly the case when the test solution is insufficiently mixed or the amount of solution (for example the size of the drop of blood) is not appropriate.
- Another disadvantage is that the card is not archivable because, on the one hand, the image of the agglutinate or absence of agglutinate is not stable over time and secondly the liquid on the surface of this card may tip over.
- test device operating on the principle of immunofiltration.
- a test device operating on the principle of immunofiltration.
- Such a device is in the form of a housing comprising, on the one hand, a reservoir for receiving the test solution and, on the other hand, a filtering membrane constituting the bottom of this reservoir, on which are deposited (in different places) specific antibodies, and designed to ensure this test.
- This protocol consists, in a first step, in taking a determined quantity of blood (more particularly by means of a pipette) so as, according to a second step, to dilute the blood taken by adding a determined quantity of diluent solution (in particular physiological saline), usually by predilution inside a suitable additional container.
- This protocol consists, finally and in a third step, in taking a given quantity of solution (blood + diluent solution) inside this container and depositing it inside the reservoir and on the filtering membrane of the test device. .
- the present invention is intended to overcome the disadvantages of devices of the state of the art.
- the invention relates to a device for detecting at least one element contained in a solution at least partly constituted by at least one component of the blood, in particular a patient, this device comprising: a reception zone of the solution at which a means for receiving this solution is defined;
- a detection zone distinct from the reception zone, and at which a means is defined for detecting the element contained in this solution; means for supplying the detection zone in solution contained in the reception zone;
- a housing at which the receiving zone, the detection zone and the feeding means are defined; means for regulating the supply of the detection zone in solution contained in the reception zone.
- This device is characterized in that the receiving zone is situated upstream of the detection zone and the receiving means is constituted by a reservoir comprising a wall at the level of which is provided a notch, constituting the means for feeding the detection zone, and arranged upstream of the detection zone, this to allow the flow of the solution contained in the receiving means of the receiving zone to the detection zone.
- this device further comprises means for regulating the supply of the detection zone in solution contained in the receiving zone.
- the detection means comprises means for ensuring filtration, more particularly an immunofiltration, the solution from the receiving means.
- the means for ensuring an immunofiltration is, at least in part, constituted by an immunofiltration membrane comprising, on the one hand, a filtration membrane, more particularly a nitrocellulose membrane or the like, and, on the other hand means, associated with the filtration membrane, and adapted to fix the element to be detected.
- the invention relates, in fact and more particularly, to a device having the aforementioned characteristics and designed for the detection of a naturally colored element contained in a solution at least partly constituted by a blood sample of a patient.
- the invention also relates to a system for evaluating the compatibility between, on the one hand, a blood solution that can be administered by infusion to a patient and, on the other hand, the blood group of this patient, system comprising two detection devices having the above characteristics.
- the invention relates to a device having the above-mentioned characteristics and designed for the detection of a naturally non-colored, weakly colored or weakly contrasted element contained in a solution at least partly constituted, either by a blood sample of a patient or by serum contained in the blood of a patient.
- this device further comprises, and at the level of the receiving means, means for binding to the element to be detected and designed to allow visual, magnetic or fluorimetric detection, at the level of the detection zone.
- the solution, containing the element to be detected is contained in the reception means defined at a receiving zone while the detection of this element is carried out by a detection means. defined in a separate detection area of this reception zone, but supplied with solution by means of a feeding means.
- This also makes it possible, advantageously, to deposit a sample containing an element to be detected directly in the reception zone (in particular by depositing, in the reception means which this reception zone comprises, a drop of blood pearling at the finger of the a patient, for example by direct finger contact with this receiving means), for, then (and if necessary), adding a diluent solution (for example physiological saline) to this sample in order to ensure dilution of this sample , this before feeding the detection means in solution constituted by the diluted sample and containing the element to be detected.
- a diluent solution for example physiological saline
- Such a procedure advantageously makes it possible to ensure dilution of the sample containing the element to be detected directly at the level of the detection device according to the invention, this avoiding a predilution of this sample in an additional container and deported.
- the dilution of the sample in the receiving means (and out of the detection means) advantageously makes it possible to avoid an immediate saturation of this detection means.
- Another advantage is that it is possible to detect an element contained in a solution made, either from a particularly limited amount (a few microliters suffice) of sample (especially blood or serum), or from a larger amount (up to at least 100 microliters) of such a sample.
- the use of the device according to the invention makes it possible to deposit a sample in the receiving zone without necessarily controlling, in a precise manner, the volume of this sample to then dilute this sample with a diluent solution (again with a quantity of diluent solution that it is not necessary to control accurately), this before feeding the detection means in solution constituted by the diluted sample and containing the element to be detected.
- Such a procedure advantageously makes it possible to dispense with the use of a pipette to carry out the detection of an element contained in a blood solution, particularly in the context of the determination of the blood group of a sample of blood.
- the receiving zone is located upstream of the detection zone and that the feeding means is constituted by a recess formed at the wall of the receiving means that comprises this receiving zone.
- the supply of the detection means that comprises the detection zone is then advantageously ensured by a flow of the solution contained in the receiving means of the reception zone towards the detection zone, this by gravity and through said notch.
- the device comprises means for regulating the supply of the detection zone in solution containing the element to be detected.
- a control means advantageously makes it possible to feed the detection zone with an appropriate quantity of solution for the smooth running of this detection and, more particularly, to avoid too rapid saturation of the detection means.
- the presence of this control means allows, advantageously and in the presence of a large amount of solution contained in the receiving means, on the one hand, to retain this solution (more particularly the red blood cells contained in this solution) in the receiving means and, on the other hand, to release, in particular periodically, prolonged or deferred, a sufficient amount of solution to carry out the detection.
- the device according to the invention advantageously makes it possible to detect an element contained in a solution at least partly constituted by at least one component of the blood, this also for very small amounts of solution and / or sample only for very large amounts of solution and / or sample.
- the device according to the invention is in the form of a compact housing particularly convenient for use.
- FIG. 1 is a schematic perspective view of a device according to the invention.
- Figure 2 is a schematic and longitudinal section of the device shown in Figure 1;
- Figure 2a is a view similar to Figure 2 and corresponds to a variant of the device shown in Figure 2;
- Figures 3, 3a, 3b are schematic representations of the device illustrated in Figures 1 and 2, this for the 3 essential steps of the protocol of use of the device according to the invention;
- FIG. 4 is a schematic view of a system for evaluating the compatibility between a blood solution to be administered to a patient and the blood group thereof;
- FIG. 5 is a schematic and perspective view of the system illustrated in FIG. 4.
- the present invention relates to the field of manufacturing devices designed to detect at least one element contained in a solution at least partly constituted by at least one component of the blood, in particular a patient.
- a solution is, more particularly, constituted by a sample (pure or diluted) of blood of a patient or by serum (pure or diluted) contained in this blood.
- such a device 1 is in the form of a housing 2 (preferentially made of plastic material) comprising, on the one hand, an area 3 for receiving the solution containing the element to be detected, on the other hand, an area 4 (distinct from the receiving zone 3) for the detection of the element contained in this solution and, secondly, a means 5 for supplying the detection zone 4 in solution contained in the receiving zone 3.
- the latter comprises a means 31 for receiving the solution containing the element to be detected.
- this receiving means 31 is designed to receive at least one sample of blood or serum that may constitute the solution containing the element to be detected.
- This receiving means 31 is, again, designed to receive a diluent solution (in particular physiological saline) that should be added to such a sample for the realization, by dilution in this means of reception 31, of such a solution. .
- said receiving means 31 is advantageously designed (in particular shaped) so as to allow a finger (more particularly that of a patient at the level of which a drop of blood constituting the sample containing the element to be detected), in particular at the end of this finger, to be introduced inside this reception means 31.
- this receiving means 31 takes the form of a reservoir 310 having a bottom 311 and delimited by at least one side wall 312 extending from the bottom 311.
- the device 1 has a substantially planar surface P defined at the housing 2 of the device 1. As can be seen more particularly in FIG. 2a, it is, in particular, at the level of this flat surface P that can be defined the bottom 311 of the tank 310 of the receiving means 31 while the side wall 312 of the tank 310 protrudes (more particularly upwards) from this flat surface P and has substantially the shape of a dike or the like.
- the device 1 comprises, more particularly at the receiving zone 3, a means 32 for regulating the supply of the detection zone 4 in solution containing the element to be detected.
- this regulation means 32 is defined at the receiving means 31, in particular (and preferably) at the bottom 311 of this receiving means 31.
- this means 32 for regulating the supply is constituted by a means designed able to hold the solution (more particularly the element to be detected contained in this solution) in the means 31. reception.
- a second characteristic of this control means 32 is that it consists of a means designed to release a quantity (which, by design of the device and / or depending on its use, can be determined or variable) solution contained in the receiving means 31, this periodically, extended or deferred.
- this quantity can, then and according to the case, correspond to both a low test portion (less than 10 microliters of solution) and a test sample. medium or high (up to at least 100 microliters).
- the regulating means 32 the latter 32 is, in fact, constituted by an absorbent support, in particular of textile type, contained in the bottom 311 of the receiving means 31.
- such a textile absorbent support is constituted by a felt or the like.
- the aforementioned reception means 31 is then designed (in particular shaped) so as to allow a finger, in particular at the end of this finger (more particularly at the level of which a drop of blood beads constituting the sample containing the element to be detected), to come into contact with the regulating means 32, in particular constituted by such an absorbent support.
- the device 1 comprises a means
- this supply means being able to adopt different forms, in particular related to the structure of this device 1.
- the device 1 comprises a housing 2 at which the receiving zone 3 is located upstream of the detection zone 4.
- This detection zone 4 is therefore located at a level lower than that of the reception zone 3 and / or below this reception zone 3.
- this detection zone 4 is at least partially defined by a cavity 42 extending below and / or below the level of the plane surface P that comprises the device 1.
- This cavity 42 is delimited by edges extend in a reentrant manner (more particularly downwards) from this flat surface P.
- the means 5 for supplying this detection zone 4 may advantageously be constituted by a means for allowing the flow (more particularly by gravity) of the solution contained in the means 31 for receiving the reception zone 3 towards the detection zone 4.
- this supply means 5 consists of a notch 51 formed at the wall 312 of the reservoir 310 of the receiving means 31, more particularly of the partition wall 312 between the zone 3 of the reception area and zone 4 detection.
- This notch 51 is, in fact, positioned upstream of the detection zone 4 as can be seen in FIGS. 1, 2 and 2a.
- Such a supply means 5 may, again, be constituted by a channel 52 connecting the reception zone 3 to the detection zone 4 (situated below this reception zone 3), more particularly the reservoir 310 of this zone 3 and the cavity 42 of this detection zone 4.
- this channel 52 extends from this reception zone 3 (more particularly from the reservoir 310, in particular from the bottom 311 of this reservoir 310) and towards the detection 4 (more particularly towards the bottom of the cavity 42 that includes this detection zone 4).
- this channel 52 can extend substantially parallel to the flat surface P and then be substantially horizontal, or even comprise (particularly at or near the receiving means 3) a portion oriented with a given angle (more particularly at 90 °) with respect to this flat surface P
- such a channel 52 preferably adopts a slope descending towards this detection zone 4 thus facilitating the gravity flow of the solution towards the detection zone 4.
- Such a channel 52 is bordered, on both sides and longitudinally, by dikes able to contain and channel the flow of this solution.
- FIG. 2a One particular embodiment of the invention illustrated in FIG. 2a consists in the fact that such a supply means 5 is, in fact, constituted by such a channel 52 coming to complete an indentation 51 as mentioned above and extending up to the flat surface P, or even to channel 52, especially to the bottom thereof 52.
- the device 1 has a detection zone 4 comprising a means 41 for detecting an element contained in a solution.
- this detection means 41 comprises means 411 for filtering, more particularly an immunofiltration (also called immunoconcentration), of the solution coming from the receiving means 31.
- This detection means 41 further comprises a means 412 for absorbing the filtrate resulting from the filtration of the solution by the filtration means 411.
- this means 412 absorption extends below the means 411 filtration and is preferably constituted by cotton, blotting paper or the like.
- the device 1 according to the invention may also comprise a bottom (not shown) designed to receive such a means of absorption 412, to maintain it 412 below the filter means 411 and in contact therewith 411, and to be associated (in particular by clipping, gluing or engagement force) to the remainder of this device 1 (more particularly to the remaining of the housing 2 of this device 1).
- the means 411 for ensuring an immunofiltration the latter 411 is, at least in part, constituted by an immunofiltration membrane comprising, firstly, a filtration membrane.
- This filtration membrane is, more particularly, constituted by a nitrocellulose membrane or the like, and / or has pores whose diameter is greater than 8 microns.
- this immunofiltration membrane comprises a means, associated with the filtration membrane, and designed to fix the element to be detected.
- This means for fixing the element to be detected is associated with the filtration membrane by deposition, fixation, incorporation or the like.
- such a means for fixing the element to be detected is constituted by a ligand, more particularly a capture antigen or a capture antibody.
- the detection device 1 can, according to a first embodiment, be designed to detect a naturally colored element contained in a solution at least partly constituted by a sample (pure or diluted) of a patient's blood.
- Such a device 1 may then, more particularly, be designed to detect at least one colored element consisting of a red blood cell belonging to a given group or to a specific subgroup of red blood cells (groups A, B, O Rhesus or other groups minors).
- a device is preferably designed to detect a plurality of types of colored elements each corresponding to a type of red blood cell belonging to a specific group or to a specific subgroup of red blood cells.
- Such a device 1 then makes it possible to detect at least one group or at least one subgroup of red blood cells contained in such a blood sample.
- the means for fixing the element to be detected is constituted by a ligand defined by a capture antibody.
- a capture antibody may be specifically adapted to capture a type of red blood cell belonging to a particular group or to a specific subgroup of red blood cells. More particularly, such a capture antibody can then be of the antiA and / or antiB type.
- the device 1 designed to detect such a colored element is, in fact, a device 1 designed to ensure the blood grouping of red blood cells belonging to at least one group or at least one subgroup of red blood cells, this within the framework of blood group test of a blood sample.
- the invention also relates to the use of such a device 1 having the above-mentioned characteristics for the detection of at least one colored element constituted by a red blood cell belonging to a specific group or to a specific subgroup of globules. red and contained in a solution at least partly constituted by a blood sample of a patient.
- Such a device 1 may then be used to group or typify the red blood cells contained in such a solution, and will thus make it possible to detect at least one group or at least one subgroup of red blood cells to which the red blood cells contained in this solution, in order to determine the blood group of such a solution at least partly constituted by a blood sample of a patient.
- the present invention also relates to a system 6 for evaluating the compatibility between, on the one hand, a blood solution (in particular blood contained in a blood bag) that can be administered by infusion to a patient and, on the other hand, the blood group of this patient.
- a blood solution in particular blood contained in a blood bag
- such an evaluation system comprises two detection devices 1 having the characteristics described above and designed to determine a blood group.
- such a system 6 comprises, on the one hand, a first test device 1 dedicated to the determination of the blood group of the blood solution that can be administered and, on the other hand, a second test device dedicated to the determination of the blood group of a patient to whom such a blood solution is likely to be administered.
- a system 6 preferably adopts the shape of a single apparatus at which the two test devices (1; 1 ') are defined, in particular in a juxtaposed manner or (and preferably as visible in FIG. 4) substantially spaced.
- Each of these devices (1; l ') then comprises a reception zone 3 (provided with a reception means 31, a tank 310, a bottom 311, a side wall 312, a means for regulating 32), a detection zone 4 (provided with a detection means 41, a means for filtering 411, a means for absorbing 412, a cavity 42) and a supply means 5 (constituted by a notch 51, preferably completed by a channel 52), these means having the aforementioned characteristics.
- this system 6 has a flat surface P common to both test devices (1; l ').
- this system comprises, on the one hand, a first means 61 for identifying the detection device 1 corresponding to the blood solution to be administered and, on the other hand, a second means 62 to identify the detection device corresponding to the patient.
- system 6 includes means 63 provided with instructions for using this system 6.
- each device (1; 1 ') included in this system 6 is completed by a peripheral wall (64; 64') making it possible to delimit such a device (1; 1 ') and to maintain each blood solution to be tested at the same time. inside the device (1; l ') which must carry out the test.
- Such a peripheral wall (64; 64 ') projects (more particularly upwards) from the common plane surface P and is substantially in the form of a dike or the like.
- the detection device 1 is, however, in no way limited to such detection.
- the device 1 according to the invention may, again, be designed to detect a naturally non-colored element, weakly colored or weakly contrasted, contained in a solution at least partly constituted by at least one blood component.
- a solution can be constituted either by a sample (pure or diluted) of a patient's blood, either by serum (pure or diluted) contained in the blood of a patient.
- Such a device 1 may, then, more particularly be designed and used to detect a serum molecule or a type of blood cell element, in particular consisting of a lymphocyte, a monocyte, a polymorphonuclear element or other.
- the element to be detected is not colored, weakly colored or weakly contrasted, it is necessary to ensure the presence, at the level of the detection device 1, of a means designed to allow a visual, magnetic or fluorimetric detection of the element to be detected, this at the level of the detection zone 4.
- This means for the detection is, in fact, constituted by a means designed to bind to the element to be detected (referred to as connecting means in the following description) and comes in addition to the aforementioned fixing means which, in the case of a non-colored, weakly colored or weakly contrasting detecting element consists of a ligand defined by a capture antigen or a capture antibody present at the level of the immunofiltration means 411.
- such a connecting means can be added to the solution containing the element to be detected, more particularly by being injected or discharged into the receiving means 31.
- a particular embodiment consists, in fact, in incorporating such a means of binding into a solution intended to ensure the dilution of the sample of blood or serum containing the element to be detected.
- the detection device 1 which comprises, at its receiving means 31, such a connecting means, especially in dried form.
- this means designed to bind to the element to be detected is constituted by a labeled ligand.
- a labeled ligand is constituted by an antibody or an antigen coupled to either colored particles, in particular colloidal gold, latex, polystyrene or other, or to an enzyme.
- an enzyme is, more particularly, supplemented by a chromogenic means, in particular present at the level of the means 41 of detection.
- FIGS. 3, 3a and 3b show the essential steps of the protocol for using the detection device 1 according to the invention.
- such a blood sample can either come from a patient and adopt, in particular, the form of a drop of blood whose volume is between 1 and 100 microliters (such a drop of blood can be deposited in the receiving means 31), or correspond to a blood solution contained in a transfusion bag and packaged inside a segment of tubing (the blood contained in such a tubing can then be deposited in the receiving means 31, this with the aid of a tubing piercing device).
- the second step of this protocol consists in adding to this sample (of blood or serum) and inside the means of reception, a diluent solution (in particular saline solution) as illustrated in FIG. 3a, this for the realization , by dilution, of a blood solution containing an element to be detected.
- a diluent solution in particular saline solution
- the solution thus produced feeds the detection zone 4 via the means 5 of supply.
- this usage protocol can, again, be completed by a final step of adding a diluent solution on the detection zone 4 so as to eliminate more effectively the elements to be characterized (in particular the red blood cells in the case of a blood group test) and not fixed specifically on the membrane.
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR0853085A FR2931244A1 (en) | 2008-05-13 | 2008-05-13 | DEVICE FOR DETECTING AT LEAST ONE ELEMENT CONTAINED IN A BLOOD SOLUTION |
PCT/FR2009/050882 WO2009147352A2 (en) | 2008-05-13 | 2009-05-13 | Device for detecting at least one element contained in a blood solution |
Publications (1)
Publication Number | Publication Date |
---|---|
EP2277052A2 true EP2277052A2 (en) | 2011-01-26 |
Family
ID=40184849
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP09757728A Withdrawn EP2277052A2 (en) | 2008-05-13 | 2009-05-13 | Device for detecting at least one element contained in a blood solution |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP2277052A2 (en) |
FR (1) | FR2931244A1 (en) |
WO (1) | WO2009147352A2 (en) |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5073484A (en) * | 1982-03-09 | 1991-12-17 | Bio-Metric Systems, Inc. | Quantitative analysis apparatus and method |
US4943522A (en) * | 1987-06-01 | 1990-07-24 | Quidel | Lateral flow, non-bibulous membrane assay protocols |
GB8903046D0 (en) * | 1989-02-10 | 1989-03-30 | Vale David R | Testing of liquids |
ES2151902T3 (en) * | 1992-03-10 | 2001-01-16 | Quidel Corp | MEANS OF SEPARATION OF RED BALLOONS TO DOSAGE THEM BY SPECIFIC UNION. |
WO1997032213A1 (en) * | 1996-03-01 | 1997-09-04 | Majesco Biologicals, Inc. | Blood testing kit |
WO2006105110A2 (en) * | 2005-03-29 | 2006-10-05 | Inverness Medical Switzerland Gmbh | Assay device and methods |
CA2658795A1 (en) * | 2006-07-26 | 2008-02-07 | Abon Biopharm (Hangzhou) Co., Ltd. | A test device for detecting an analyte in a liquid sample |
-
2008
- 2008-05-13 FR FR0853085A patent/FR2931244A1/en not_active Withdrawn
-
2009
- 2009-05-13 EP EP09757728A patent/EP2277052A2/en not_active Withdrawn
- 2009-05-13 WO PCT/FR2009/050882 patent/WO2009147352A2/en active Application Filing
Non-Patent Citations (1)
Title |
---|
See references of WO2009147352A3 * |
Also Published As
Publication number | Publication date |
---|---|
FR2931244A1 (en) | 2009-11-20 |
WO2009147352A3 (en) | 2010-03-25 |
WO2009147352A2 (en) | 2009-12-10 |
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