CN1252475C - Agglutination reaction and separating container - Google Patents

Agglutination reaction and separating container Download PDF

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Publication number
CN1252475C
CN1252475C CNB03140653XA CN03140653A CN1252475C CN 1252475 C CN1252475 C CN 1252475C CN B03140653X A CNB03140653X A CN B03140653XA CN 03140653 A CN03140653 A CN 03140653A CN 1252475 C CN1252475 C CN 1252475C
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CN
China
Prior art keywords
chamber
lining
box
plug
post
Prior art date
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Expired - Lifetime
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CNB03140653XA
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Chinese (zh)
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CN1487298A (en
Inventor
W·米尔彻尼奥斯基
M·祖列克
K·J·雷斯
D·E·比尔托尔德
L·戴维斯
T·M·塞卡维杰
D.M.戴维斯
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Ortho Clinical Diagnostics Inc
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Ortho Clinical Diagnostics Inc
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Priority claimed from US08/595,719 external-priority patent/US5780248A/en
Application filed by Ortho Clinical Diagnostics Inc filed Critical Ortho Clinical Diagnostics Inc
Publication of CN1487298A publication Critical patent/CN1487298A/en
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Publication of CN1252475C publication Critical patent/CN1252475C/en
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    • 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
    • G01N33/5302—Apparatus specially adapted for immunological test procedures
    • G01N33/5304—Reaction vessels, e.g. agglutination plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/026—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations having blocks or racks of reaction cells or cuvettes

Abstract

A vessel for conducting blood cell agglutination assays is disclosed. A barrier retains reactants in an upper chamber during incubation, then, in response to a force, permits reagents to enter a lower chamber containing a matrix for separating agglutination.

Description

Agglutinating reaction and separation container
The application is dividing an application of application number 97110916.8.The applying date of original application is on February 2nd, 1997, and the denomination of invention of original application is " agglutinating reaction and a separation container ".
The application is that application number is 08/093,106, the applying date is the part subsequent application of the U.S. Patent application on July 16th, 1993, and this piece U.S. Patent application to be application number be 08/092,157, the applying date is the subsequent application of on July 15th, 1993, existing resigned U.S. Patent application.
Technical field
The present invention relates to the agglutination assay field, relate in particular to the container that is used to carry out CA and separates agglutinator.
Blood group serology need be measured the haemocyte compatibility between blood donor and the receptor before to patient blood transfusion or organ transplant.By confirming antibody that contains among the patients serum and the compatibility of measuring cell from no immune response between the antigen in blood donor's the haemocyte.
On the erythrocytic surface of each individuality, find many different blood group antigens.Blood group is identified and is normally checked red blood cell so that measure the process whether antigen exists.Usually utilize known specific antibody to finish this process.
For the serum of checking patient or the antibody in the blood plasma, the reagent that will contain the haemocyte with known antigens mixes with blood serum sample.With reactant incubation a period of time, when the antibody at those antigens exists, be enough to make red blood cell generation aggegation during this period of time.Then that potpourri is centrifugal, if exist by the red blood cell of aggegation, so can be clearly visual to these agglutinators in the bottom of reaction vessel, so just show to have direct antibody in the sample at known antigens on the red blood cell.If there is not direct antibody in the sample at known antigens on the red blood cell, aggegation can not take place just, this shows the centrifugal red blood cell that does not have later aggegation.
Recently, worked out such system already: promptly in a part of container, carry out agglutinating reaction, a kind ofly will be separated the red blood cell of institute's aggegation from other component of reagent/specimen potpourri by the matrix that the cell of aggegation is separated and in another part of same container, utilize.U.S. Patent application No.08/407 in common unexamined, 747 and 08/112, disclose and described such system in 402, these the two pieces of document number of being application numbers are 08/023, the subsequent application of 500 United States Patent (USP) (abandoning now), the holder that these applications are generally the application has.Therefore the content of every piece of application is incorporated herein by reference.Also can be used in the aforementioned application text in the disclosed invention according to agglutinating reaction of the present invention and separation container, is by Ortho Diagnostic SystemsInc., Raritan, New Jersey, under thetrademark BIOVUE TMProduce and market.Such reaction vessel is to have a cylindricality that goes up a chamber and a following chamber, and it is bigger than following chamber wherein to go up the diameter of chamber.Following chamber contains and is useful on the matrix that the aggegation cell is never separated in the aggegation cell.The diameter of following chamber is narrow as to be enough to when joining reagent and sample in the chamber (adding with transfer pipet typically), if do not apply extra power, reagent and sample be just as staying in the chamber, and can not enter down the chamber.
Indirect antiglobulin test (being called antiglobulin test) is one and is used for measuring the patients serum and whether has blood test at specific antigen IgG antibody on the red blood cell surface.In antiglobulin test, incubation serum under the situation that has red blood cell reagent is so that combine the lip-deep antigen of antibody and red blood cell.These IgG antibody itself can make erythrocyte agglutination hardly, and perhaps only aggegation a little is not enough to use the routine techniques sight check.Usually need to add second antibody, to be convenient to obtain observable aggegation at human IgG.
In the red blood cell somatotype, a blood test is used to measure the red blood cell surface and whether has specific antigen, red blood cell to be analyzed is joined in the chamber, with an after-applied power, centrifugal force for example enters red blood cell to contain the following chamber at the antibody and the isolation medium of specific red blood cell antigen.If red blood cell has the antigen that combines with specific antibody in the following chamber on its surface, then form agglutinator, and separate agglutinator with matrix.In the blood measuring method of other type (for example reverse somatotype), be measure among the patients serum directly aggegation, at the antibody of red blood cell antigen, the patients serum is joined in the chamber with the red blood cell reagent that its surface has known antigens, and apply a power (for example centrifugal force), so that reactant enters the following chamber of containing liquid medium and isolation medium but not having antibody.In this measured, the antibody of existing direct aggegation produced the agglutinator that utilizes matrix to separate among the patients serum.
In the blood measuring of another kind of type, make to have known specific antibody reagent at red blood cell antigen and be deposited in the chamber with patient's red blood cell.If antibody reagent is direct agglutinating antibody, then apply a power (for example centrifugal force), and do not have former incubation process, inclusions is forced into down in the chamber, and the isolation medium that is in the aqueous solution is contained in following chamber.Separate agglutinator with matrix then.In addition, also can make patient's red blood cell be deposited in the chamber and add and have known specific IgG antibody reagent, carry out incubation subsequently, so that make antibodies to the red blood cell surface on the predetermined antigen.After the incubation, apply a power (for example centrifugal force), so that make reactant enter down the chamber, following chamber is contained isolation medium and is specific to the anti--IgG antibody of the erythrocytic IgG antibody reagent that is used for chamber on the incubation.If antibody reagent is present on patient's the cell surface, then descend the anti--IgG antibody in the chamber to be easy to form the agglutinator that utilizes matrix to separate.
With sample and one section time enough of reagent incubation, so that take place direct aggegation (as in) or antibody-antigen-reactive (as in antiglobulin test) in cell typing test afterwards by centrifugal to the reaction vessel pressurization, thereby make reactant be pressed against the bottom of post and on isolation medium.Centrifugal result is, UA material moves downwards by isolation medium, and the cell of institute's aggegation is retained in the top of isolation medium or distribute in the substrate according to the aggegation degree.Stronger agglutinating reaction causes cell to be retained on the upper position near isolation medium, and more weak agglutinating reaction cause agglutinator be distributed in the position that has various different distance above the matrix on.
Why to be retained in the top of post be the capillary result who passes the termination of post bottom for sample and reagent between incubation period, and the diameter of its center pillar bottom reduces with respect to top.Two potential error sources when having pointed out already to measure with this post.At first, if with the power of transition reagent and sample are directly dripped to the center of reaction chamber, then reactant can directly be deposited in down above the isolation medium in the chamber and not be retained between incubation period in the chamber.Like this, reactant began to enter isolation medium before finishing aggegation.Secondly, the thinning agent or the solution that contain isolation medium might enter the chamber.For example when transporting or handle container, produce this error by spilling or other Recombinant Interferon.When the solution that contains isolation medium or thinning agent also had antibody or other directly to influence the reagent of test findings, such spilling meeting caused using some the reagent coupled columns from other post to cause cross pollution.This phenomenon occurs in when the user inserts the transfer pipet tip reaction chamber, the reagent contamination of spilling tip, this tip is transferred in another container by transfer pipet again then.This can cause the error result in the CA.
Summary of the invention
Therefore, an object of the present invention is to provide a kind of improved mechanism that between the incubation period of CA, keeps sample to separate with reagent.A further object of the present invention provides the device that is used for preventing the material displacement that contains the post bottom.
The invention provides a kind of improved container that is used to carry out agglutinating reaction and separates agglutinator.This container comprises that one is held the last chamber of reactant, the following chamber of a separation agglutinator and the barrier device of a split cavity, this barrier is before the inclusions that will go up the chamber is incorporated on the isolation medium, reactant is retained in the chamber, and work as a power (promptly, greater than atmospheric pressure) when being applied on the barrier, the inclusions of chamber is moved to down the chamber from last chamber.In a preferred embodiment, this barrier is comprising a narrow path between the chamber up and down.The plug-in unit of plug-in unit, crimping or the spirality by will having slot or other similar geometric casting during manufacture or insert up and down and can form narrow gap between the chamber.This plug-in unit provides a physical barriers that can reduce the hole dimension between following, the following chamber, thereby avoids down the inclusions of chamber to pollute upward chamber.And plug-in unit also makes sample and reagent separate when incubation.When plug-in unit was spirality, the path that is limited by screw, central shaft and the post jamb of spiral was little as to be enough to be retained in the chamber at normal gravity and atmospheric pressure ground next time liquid.For example, the diameter of spirality plug-in unit is in the scope of about 0.110-0.140 inch.The diameter of spirality inserter shaft is approximately the 0.030-0.090 inch.The nearly 6-30 of a spirality plug-in unit per inch screw thread.
Carry out ultra-sonic welded by coupled columns, load post with reagent subsequently and also can form narrow gap.Path also can comprise a shelf-shaped barrier.In the present embodiment, dividing plate has little a center pit that is enough under normal gravity and atmospheric pressure fluid is retained in the chamber.Dividing plate is silicon rubber preferably.Path also can comprise the barrier of a porous stopper.Porous plug has the path that passes it, and these paths are little as to be enough under normal gravity and atmospheric pressure fluid is retained in the chamber.Porous plug is preferably poly.
In another embodiment, the present invention includes a lining, this lining is to have a conical part form of passing the hole of narrow tip.Before moving on to the sample suction in the container, this lining is placed on the tip and stretches into the reaction vessel post, thereby the cross pollution of having avoided thinning agent in the container or solution to enter another container and having caused, otherwise another container may be by cross pollution in moving the liquid process.The final user is inserted into described lining the top of post expediently before suction moves reagent.Lining has a unit form in six conical parts that are arranged side by side or pond.The total area of lining and shape and BIOVUE TMThe top side of box is identical.
Lining comprises a main body and at least one conical part that stretches out from main body, wherein said conical part has a hole on its narrow top, and described conical part with described narrow top spaced positions on packoff is arranged.Conical part comprise one spaced apart and have first end of a perforation and one and isolated second end of described first end on its narrow top with described main body, wherein said packoff is positioned on second end or near second end.Packoff comprises an O shape ring around described conical part, and O shape ring can be connected as a single entity with conical part.In a preferred embodiment, lining has six linearly conical parts that stretch out on the main body, and the reaction vessel of the size of lining and box is complementary.Lining is preferably acrylic acid.
The present invention further imagination utilize a metal foil seal, be included in the box that wherein is linearly aligned six reaction vessels, and one comprise that a main body and six are the lining of the conical part that linearity stretches out from it, and wherein conical part comprises a narrow tip that is suitable for passing described metal foil seal.And at this moment this lining of Cha Ruing frictionally is engaged in the described box, so that the bonding part between seal box and the lining.The device that is used for sealed engagement part be around the O shape ring of each conical part and preferably and each conical part be integrated.
Description of drawings
Fig. 1 shows reaction and the separation container with a plug-in unit, and this plug-in unit has a narrow perforation in last reaction chamber.
Fig. 2 is the top view that the box of six reaction vessels is arranged, and wherein storage container does not have plug-in unit, and a container (left side is played second) contains a plug-in unit as shown in Figure 1, and a container (left side is played the 3rd) contains reactant.
Fig. 3 is the sectional view along the box with reaction vessel of the 3-3 line of Fig. 2.
Fig. 4 is the side view with box of reaction vessel.
Fig. 5 is the sectional view along the 5-5 line of Fig. 2, shows an indoor plug-in unit with a narrow perforation on reaction vessel.
Fig. 6 shows the last chamber of the reaction vessel with a narrow perforation.
Fig. 7 shows the plug-in unit with an extension, and this extension has a perforation in the chamber down.
Fig. 8 is the sectional view along the 8-8 line of Fig. 3.
Fig. 9 shows reaction and the separation container that has been crimped under last chamber.
Figure 10 is the sectional view along the 10-10 line of Fig. 9.
Figure 11 has the reaction of a spirality plug-in unit 1 and the side view of separation container in last chamber 2, the bottom 3 of last chamber has been modified, is used to hold described plug-in unit.
Figure 12 is the side view of the structure of spirality plug-in unit; (A) showing overall diameter 1 is 0.120 inch and interior shaft diameter 2 is 0.060 inch a plug-in unit; (B) showing overall diameter 1 is 0.120 inch and interior shaft diameter 2 is 0.08 inch a plug-in unit.
Figure 13 is that overall diameter 1 is 0.120 inch and interior shaft diameter is the side view of 0.06 inch spirality plug-in unit.
Figure 14 shows a pond of the lining in the last chamber of the reaction vessel post of packing into.Has O shape ring 1 around the pond under the lining main body of each conical part or pond.
Figure 14 A is the planimetric map in a marked price pond of the lining in a kind of last chamber of the reaction vessel post that is designed to pack into.All there is an O shape ring 1 around the pond that is positioned under the lining main body in each pond.The figure shows the narrow angle that pointed top is arranged in lining pond, design is in order to pierce through the seal of basin like this.
Figure 14 B is the top view in a pond of lining, and innermost circle further illustrates the perforation by it.
Figure 14 C is the planimetric map in a pond of real cover, shows the angle on lining top, and design is the box for puncture through seal like this.
Figure 15 shows and comprises that six have the conical part on pointed top or the lining in pond, packs in the last chamber of box with six reaction vessel posts in described pointed top.Has O shape ring 1 around the parts of each parts under the lining main body.Each parts also has tip 2 to be used to pierce through the metal foil seal of the reaction vessel post that covers box.
Figure 15 A is the top view that comprises the lining in six conical parts or pond.
Figure 15 B is the planimetric map in an independent lining pond, shows the angle of understanding the lining top that is designed to make the box of wearing sealing.
Figure 15 C is the planimetric map that comprises the lining in six conical parts with pointed top or pond, packs into by the seal that pierces through box in the last chamber of box with six reaction vessel posts in described pointed top.
According to the present invention, will the container that be used to carry out agglutinating reaction and separate agglutinator be described with various embodiment form.By to agglutinating reaction with by Ortho DiagnosticSystems Inc., Raritan, New Jersey, under thd trademark BIOVUE TMThe description of the box-like separation container of production and selling can be expressly understood some embodiment of the present invention.
Container of the present invention can be made with any appropriate materials (for example glass or various plastics) of agglutinating reaction or separation and visual result of not disturbing.In a preferred embodiment, container is made by polypropylene.
But the last chamber Any shape and the size of container are used for holding reagent and sample when carrying out incubation.Typically, be columniform on the most of in the above parts in last chamber.Barrier between the chamber limits the following interface of chamber and the last interface of following chamber usually up and down.In a preferred embodiment, the barrier of the bottom of chamber is taper in the formation, and its top stretches to or stretch into down the chamber, shown in Fig. 1,5,6,7.The part of barrier is used for keeping the reagent and the sample of chamber under normal gravity and the atmospheric pressure conditions between incubation period, and when applying a power (for example increased pressure or centrifugal force), can make fluid flow to second Room from first Room.This can pass through various devices, and for example the combination of aperture, film, piston, obturator, the plug-in unit with any geometric configuration or filter screen and any of these device is finished.In a preferred embodiment, barrier comprises a hole, and the diameter in this hole is little as to be enough to avoid under normal gravity or atmospheric pressure fluid to flow to second Room from first Room, and under increased pressure fluid is flow through.Hole 1 is positioned at the top of the tapering part of chamber, both can also can contain at (shown in Fig. 1,5 or 7) in the plug-in unit 2 to be one (as shown in Figure 6) in last chamber.
The diameter of aperture little be enough to make the fluid in the chamber surface tension avoid under normal gravity or the atmospheric pressure fluid from the chamber flow to chamber down, and under increased pressure or gravity, can overcome surface tension, so inclusions flows to down the chamber from last chamber.Change the diameter in hole according to the size of being exerted oneself, that is, bore dia is less when applying bigger power, and bore dia is bigger when applying less power.Diameter can also change, so that adapt to particles with different sizes in the reagent.In a preferred embodiment, the diameter in hole is in the scope of about 0.010-0.050 inch.In a particularly preferred embodiment, the diameter in hole is 0.020 inch.
In another embodiment, separating up and down, the barrier device of chamber comprises a spirality plug-in unit; But this spiral screw-shaped structure.Though it is circular or columniform that also available elliptical configuration, spiral are preferably on the diameter.Be similar to aperture described above, by axle, the screw thread of spiral, and its diameter of path of limiting of locular wall little be enough to stop under normal gravity or the atmospheric pressure fluid from the chamber flow to chamber down, and fluid is flowed.Another effect of plug-in unit is to have reduced thinning agent or solution to pollute the possibility that goes up the chamber from chamber spilling down thus to last chamber.This spilling can take place transporting with operating period for example.Spiral in last chamber, be positioned at as shown in figure 11 on the bottom 3 of chamber, perhaps be cast into one in this position and reaction vessel.
In the situation of chamber, available any appropriate materials (for example glass or plastics) of agglutinating reaction or separation or visual result of not disturbing is cast plug-in unit on the spirality plug-in unit individually is inserted into.Material is plastics preferably, for example polypropylene, polyamide (for example nylon), acetate resin (Delrin for example TMOr Delrin P TM), crosslinked tygon/divinylbenzene (Rexolit-e for example TM), poly-carbonic acid is tygon.In a preferred embodiment, material is a polypropylene.
Spiral is any such geometric configuration: promptly, the surface tension that makes fluid in the chamber stop under normal gravity or the atmospheric pressure fluid from the chamber flow under the chamber, and under increased pressure or gravity, can overcome surface tension again, so inclusions flows into chamber down from last chamber.For example, the angle of pitch of screw is any such angle: promptly, can make fluid (for example containing blood cell) flow to chamber down from last chamber under increased pressure, avoid simultaneously from the fluid or the matrix of chamber are polluted upward chamber down.Can change the angle of pitch according to the size of being exerted oneself, promptly when applying bigger power by screw axis and screw thread and locular wall limit regional less, and this zone is bigger when applying less power.Also can change the geometric configuration of spiral, so that adapt to particles with different sizes in the reagent.And, plug-in unit strengthened avoid post inclusions (containing isolation medium and thinning agent or solution) spilling to last indoor possibility.
In the embodiment of spirality plug-in unit of the present invention, only stop the effect (low side of numerical range) that last chamber polluted from the fluid of chamber down and the ability of sample (for example red blood cell) by spiral (numerical range high-end) can limit per inch under increased pressure the screw thread number and the depth of thread with this spiral.Because previous reasons, for make red blood cell by barrier device and other agglutinator be easy to by, preferably make limit path screw thread, central shaft and post jamb on quality and final stage comparatively level and smooth.In the face of the wall of the threaded portion of the spirality plug-in unit of post jamb point or flat.
In a preferred embodiment, spiral is for the BIOVUE of the present universal architecture that is used to comprise six reaction vessels TMIn the box, the nearly 6-30 of its a per inch screw thread, the nearly 12-20 of a per inch screw thread more preferably, be approximately 0.033 from a threaded top in about 0.166 inch scope to the following pitch of measuring than the top of low thread, more preferably be approximately 0.050 to about 0.083 inch.
The overall diameter of spirality plug-in unit arrives in about 0.140 inch scope about 0.110, more preferably is approximately 0.120 to about 0.130 inch.
The spirality inserter shaft arrives in about 0.090 inch scope about 0.030, more preferably is approximately 0.060 to about 0.080 inch.Then with reference to shown in Figure 11,12 and 13, these several width of cloth illustrate the relative geometric configuration of insertion parts.With reference to Figure 13, the following approximate geometry (unit: inch) that shows:
Threads/inch DIM3 DIM6
12 14 16 0.083 0.070 0.060 0.020 0.020 0.015
As the improvement of putting into last chamber as shown in figure 11 a kind of alternative of round diameter spirality plug-in unit of bottom, can also be with the oval spiral of pitch at this with similar geometric.As the description of front to round diameter spirality plug-in unit, oval barrier device is the spirality obturator that also is cast into a unit with the chamber of a card format both.
Replace as another, can carry out ultrasonic bond by coupled columns after the post that reagent is packed into and fetch the formation pinch path of asking of chamber up and down.In addition, barrier can comprise dish or the piston that a porosint that is positioned at the top of following chamber or post is made.Piston is columniform, and between the chamber, inclusions (for example reagent and the isolation medium) spilling of chamber was in last chamber under being suitable for stoping about its big I was installed in.Perforated piston also be suitable for preventing sample prematurely (for example centrifugal before) pass through, and should be that sample (for example centrifugal down) under the pressure greater than atmospheric pressure passes through piston.Perforated piston can not disturb agglutinating reaction or separation or any visual result or makes with the material that wherein any component does not have specificity to combine with any, and these examples of material have glass, and particularly scintillation glass also has plastics (for example polypropylene).
Other suitable barrier has solid flange, the bench formula grid that centers on chamber interior walls or other similar distortion path in spiral, and these barrier forms all have inclusions contaminated effect in chamber in the foregoing prevention.Barrier also is placed on ring valve or the dividing plate in the post.This dividing plate for example can have a center pit or perforation, and radially rules for from the post jamb to the central hole dividing plate.When applying power, dividing plate is opened, and makes the sample inclusions flow into chamber down from last chamber.Dividing plate can be made with any appropriate materials of agglutinating reaction or separation or any visual result of not disturbing.Dividing plate can be made with any suitable pliable and tough plastic material (for example silicon rubber).
When container of the present invention is used to carry out agglutinating reaction and separate, reagent and sample is joined carry out incubation in the chamber.When barrier carries out incubation sample and reagent are retained in the chamber.In a preferred embodiment, barrier has a hole, perhaps barrier is the spirality card format, and the diameter in hole or the geometric configuration of spiral are respectively little that the surface tension that is enough to make liquid and sample pass hole or spiral is retained in inclusions in the chamber under normal gravity and atmospheric pressure.Enough incubations are after the time, utilize any different mode (for example centrifugal, pressurization or absorb) basically along axially applying a power to barrier from last chamber to chamber down.This power must be enough to overcome barrier and make the inclusions in the chamber enter down the chamber.In a preferred embodiment, barrier comprises a hole, and perhaps barrier device comprises a spirality plug-in unit, and surface tension can be made every effort to overcome clothes, and inclusions enters down the chamber from last chamber and flows to the isolation medium.
The important part of barrier is that also it has reduced thinning agent in time chamber or the solution possibility to last chamber contamination, otherwise, for example transporting with operating period because this pollution can take place for spilling or other interference.
Another embodiment of the present invention is a lining that can insert the reaction vessel top before adding sample.This lining comprises a main body and one or preferably a plurality of parts or pond of stretching out from main body, and this pond is taper or funnel shaped.When the narrow top in the pond that will contain a hole was inserted in the box, it was towards the inner orientation of container, shown in Figure 14 and 15.Lining is applied to or does not have in the container of barrier device (for example crimping or plug-in unit).In a preferred embodiment, lining comprises a pond with a perforation, 1. the diameter in this hole is little as to be enough to stop the sample that will be transported in the container, be subjected to the effect of power (for example by centrifugal) up to container, and can not prolonged the incubation time by the heat insulating function of the clearance between the isolation medium in top and the post; Perhaps 2. big must to be enough to make sample to be carried easily flow into reaction vessel indoor in this hole, and allow pilot system to contact with the spilling liquid of reagent prematurely.The pond of lining with narrow top spaced positions on preferably have a packoff under (for example its outer tip end), the lining main body.About this point can refer to figs. 14 and 15.Packoff is a bulge loop best and lining is connected as a single entity.The pond of lining is installed in the last chamber of reaction vessel, so as normal operating period the pond can not be removed easily.Bulge loop or O shape ring are installed around the edge of container at the upper surface of box, have so just sealed the bonding part between lining and the box.Bulge loop or O shape loop resistance end the capillary action of post inclusions from a post to another post of any spilling.
As mentioned above, when plug-in unit not being installed in the container, thinning agent or the solution that contains isolation medium are transporting or operating period can enter the chamber.In this case, isolation medium also contains antibody or other directly influences the reagent of test findings, and this splatter can cause from the reagent of other post this post being caused cross pollution.When the last chamber that the user inserts reaction vessel with the transfer pipet tip, the reagent contamination of spilling tip, this cross pollution can take place when tip was transferred in another container with transfer pipet again then.And the result that the latter can lead to errors in CA.
The purpose of lining is the cross pollution that prevents that reagent from causing to next container from a container in moving the liquid process; Transport with operating period reagent can upwards be splashed in the chamber.
Lining is conical part or pond, and they are positioned at the top of each reaction vessel separately.Yet in a preferred embodiment, with reference to Figure 15, lining is a unit alone that has six conical tanks of arranging side by side, is connected as a single entity with the lining main body.This structure makes a lining unit that contains six single ponds be positioned at the BIOVUE of six reaction vessels TMThe top of box.
With reference to Figure 14 and 15, each taper pond of lining has a narrow tip, and a perforation by it is arranged on tip.Work as BIOVUE TMContainer is a box that comprises six posts, and when sealing the top of each post with metallic foil, the metal foil seal band on all six posts will be pierced through in the tip of lining under normal pressure.Then by move liquid with sample transfer in the pond.Like this, clean lining pond can not be subjected to any pollution of reagent or isolation medium, otherwise reagent or isolation medium can contact sample, are brought to again in another pillar then.
Final user's manual manipulation or utilize one the band prong instrument lining can be inserted in the post expediently.For the lining with six ponds inserts BIOVUE TMIn the box, wear the metal forming of box and insert in the box fully by shaking lining with the lining spine.Because the area of lining main body is identical with the top surface of box with shape, so lining can be held in place in the process of operating case easily.Use by this way lining can the interference measurement performance or result's (for example: centrifugal, UA red blood cell enters in the post by the free path of isolation medium, the red blood cell of aggegation) yet, can not stop the cross pollution of reagent coupled columns during use with the post of O shape ring.Referring to the band O shape ring of example 10 and 11 and the contrast performance test of not encircling with O shape.Result herein confirms that O shape ring can stop the cross pollution of reagent coupled columns, and this pollution is because the metal forming of box and the reagent between the lining " siphon " act on, and consequently reagent flows by the top of box.
Available any appropriate materials (for example glass or plastics) of agglutinating reaction or separation of not disturbing is made lining and its pond.Material must be suitable for piercing through metal on the box seal that spreads out.In order effectively sample to be transferred to from the pond of lining in the chamber, the pool wall of lining smoother all on quality and in the finished product preferably.Material is for example polyester, acetal, acrylic acid, acrylic nitrile-butadiene-benzene (ABS), nylon, polycarbonate, polyamide or polypropylene of plastics preferably.In a preferred embodiment, material is an acrylic acid.
At BIOVUE TMIn the system, 10 μ l red blood cell reagent are joined among the 40 μ l patients serums and, so that the patient IgG antibodies that makes prediction is to erythrocyte surface antigen with the low ion temperature solution of 40 μ l incubation 10 minutes in last chamber.Separately add these and measure composition, and importantly, it is retained in the chamber,, thereby make the ratio of low ionic-strength solution and red blood cell and serum in the mensuration process, keep constant so that mix.Barrier is easy to accomplish this point under normal gravity and pressure.It also can reduce during application of sample any mensuration composition and be forced into probability in the chamber down.Barrier can also make the mensuration composition be retained in the chamber between whole incubation period.
The important part of barrier also be to stop anti-human immunoglobulin(HIg) (IgG) antibody be attached to prematurely on the anti erythrocyte antibody of prediction before red blood cell combines, thereby reduced the aggegation probability that occurs in down the chamber at last.After the incubation, apply centrifugal force, make the inclusions of chamber enter down the chamber by barrier, the anti-human immunoglobulin(HIg) (IgG) that combines with the lip-deep patient's immunoglobulin (Ig) of red blood cell reagent (IgG) is contained in following chamber, thereby the formation agglutinator, agglutinator can not flow to down the bottom of chamber by matrix.
Embodiment
Following example is just in order to understand the present invention, rather than limiting the scope of the invention.
Example 1
The BIOVUE that will have plug-in unit TMPost compares with the post that does not have a plug-in unit, so that measure every kind of structure is used to keep space air bound that reactant and isolation medium are separated between incubation period effect.Utilization has the plug-in unit in 0.040 inch hole.40 microlitre damping fluids are joined in each pillar of 840 pillars that are used for testing.The angle that approximately is 45 ° with the Z-axis with post is used transfer pipet artificially, so that carry this 40 microlitre solution.Observe post then, whether remain on the airspace under the assaying reaction chamber.Table 1 has provided the number of " spilling ".
Table 1
The experiment number Spill number Spill number percent
The pillar that has plug-in unit 840 0 0％
Not with the pillar of plug-in unit 840 231 27.5％
Example 2
Also reagent is joined in post (have and do not have plug-in unit) and 37 ℃ of following incubations 10 minutes.Each root pillar to 480 pillars that are used for testing adds 40 microlitre buffering agents, 40 microlitre serum and 10 microlitre red blood cell suspensions.Use transfer pipet with about 45 ° angle, so that pipette reactant.Incubation is checked pillar, so that whether maintain the airspace under the assaying reaction chamber after a period of time.Table 2 has provided " spilling " frequency.
Table 2
The experiment number Spill number Spill number percent
The pillar that has plug-in unit 840 0 0％
Not with the pillar of plug-in unit 840 16 3.3％
Example 3
With aupette 40 microlitre buffering agents are charged into pillar with about 45 ° angle.Typically used than the manual mode of operation feeding force of aupette is big.Observe after filling, to be determined at whether maintain the airspace under the reaction chamber.Table 3 has provided the result of the pillar of the pillar that has plug-in unit and no plug-in unit.
Table 3
The experiment number Spill number Spill number percent
The pillar that has plug-in unit 240 0 0％
Not with the pillar of plug-in unit 240 103 43％
Example 4
Vertically 40 microlitre buffering agents are charged in 240 pillars with a transfer pipet.Owing to vertically hold transfer pipet, so liquid stream is with bigger pressure compression perforation, so probably break through the airspace that reaction chamber and separation chamber are separated.Table 4 has provided the result of test this time.
Table 4
The experiment number Spill number Spill number percent
The pillar that has plug-in unit 240 0 0％
Not with the pillar of plug-in unit 240 144 60％
Example 5
Utilize aupette vertically 40 microlitre buffering agents also to be charged in the reaction chamber of 240 pillars, more likely destroy following airspace than holding aupette like this with an angle.Table 5 has provided and has utilized the result who has the pillar of plug-in unit and do not carry out these tests with the pillar of plug-in unit.
Table 5
The experiment number Spill number Spill number percent
The pillar that has plug-in unit 240 0 0％
Not with the pillar of plug-in unit 240 204 85％
Effect of the present invention is, except the airspace that can keep between the incubation period of test between reaction chamber and the isolation medium, also can be used as the device that prevents spilling, this spilling can take place transporting, cause down the part inclusions of separation chamber upwards to be splashed in the reaction chamber with operating period.In order to test the validity that prevents spilling, will have the box of plug-in unit and not be transported to markon Fu Niya from the New Jersey, and then transport back with the box of plug-in unit.Transport by aviation and land, this process comprises loading, unloads and delivers to the laboratory.After transporting back and forth, check box, whether the liquid of spilling is arranged in the reaction chamber.The results are shown in Table 6.
Table 6
The experiment number The pillar number that the liquid spilling is arranged The number percent that the pillar of liquid spilling is arranged
The pillar that has plug-in unit 816 30 3.7％
Not with the pillar of plug-in unit 768 571 74.3％
Example 7
Carry out another and transport research,, can reduce the possibility of spilling so that test utilizes the plug-in unit with hole that size reduces.Opening diameter between reaction chamber and the isolation medium is 0.025,0.020 and 0.015 inch.Fill 600 pillars with each plug-in unit.Control group is provided with plug-in unit.The packing box also carries out indoor substitute and transports research, that is, box is fallen down 10 times from 3 meters high places.The angle of control box is so that drop on all it six planes and angle and 3 limits container.Destruction situation during this standardization experiment representative is transported and operated.Inverse relation between the size of the indication window as a result that provides in the table 7 and spilling reduce.
Table 7
The experiment number The pillar number that the liquid spilling is arranged The number percent that the pillar of liquid spilling is arranged
Post with 0.015 plug-in unit 600 75 13％
Post with 0.020 plug-in unit 600 120 20％
Post with 0.025 plug-in unit 600 132 22％
The post of no plug-in unit 600 600 100％
Example 8
Another kind of mode is can reduce aperture between reaction chamber and the following separation chamber by " curling " box.Utilize punching press can realize this point, wherein exactly the box neck under reaction chamber is impacted.The degree that power of impacting and duration decision opening are reduced.The shape of the shape decision opening of percussion tool.It is available that several structures are arranged.Can on production line, finish curly course after in post that reagent and beaded glass are packed into.
Be curled from 816 pillars on the production line, thereby as mentioned above, but the opening between defined reaction chamber and the isolation medium.This curling causes passing the section shape (as shown in figure 10) in a represented zone of bracket among Fig. 9.The control group post that these posts and 768 are curled is packaged together and be transported to markon Fu Niya, and then transports (as previously mentioned) back from that.Table 8 has provided the result that the liquid spilling that caused by shipping conditions reduces to the possibility in the reaction chamber.
Table 8
The experiment number The pillar number that the liquid spilling is arranged The number percent that the pillar of liquid spilling is arranged
The pillar that crimping is arranged 816 548 67％
The pillar of no crimping 768 571 74％
Example 9
Utilize spirality plug-in unit barrier device come defined reaction (on) chamber and the orifice size that separates between (descending) chamber.The spirality plug-in unit exactly is cast into and is inserted by polypropylene on improved in the box neck under the chamber.In manufacture process, the spirality plug-in unit is put into the box neck after in reagent and isolation medium (for example beaded glass) post of packing into.
Example 10
The BIOVUE that lining is had O shape ring TMThe post that post and lining do not have O shape ring compares, so that measure the cross pollution whether O shape ring can stop the reagent coupled columns.
By box is patted or hit on solid working surface, can imitate when transporting box and in last chamber He on the metal forming, produce spilling.
Observing lining does not have 192 boxes of O shape ring, has seen after lining is in the post that inserts box whether siphon of the reagent in the post.Half (96) lining is manually to insert in the box, and second half is to utilize two prong instruments of above-mentioned discussion to insert.
Observing lining has 336 boxes of O shape ring, looks at whether to have the siphon of reagent in the aforesaid post.Lining is manually inserted in half (168) box, and insert with two prong instruments second half (168).
Come the siphon of visual measuring column by the top side of checking box.Can measure this siphon effect by the metal forming top of observation box and the reagent fluid between the lining main body downside.
Table 9 shows number and the number percent that the sum that at every turn is used for the box tested has the box of siphon.The method that lining inserts does not influence uses the result with O shape lining papers cover, but the result of no O shape lining papers cover is used in influence.As shown, when lining manually inserts (box is on inverted position), double when approximately to be lining with instrument insert (box is on the upright position) of the number with box of siphon reagent.
Table 9
The method that lining inserts The lining of no O shape ring The lining that O shape ring is arranged
Use instrument 39/96(40.6％) 0/168
Manually 74/96(77.1％) 0/168
Example 11
The box and the lining that lining are not had O shape ring have the box of O shape ring to carry out functional trial relatively.Box is transported in simulation, produces the reagent spilling in last chamber and on the metal forming, and for example 10 is described.
As mentioned above, with two these dual modes of prong instrument of utilization lining is inserted in the post of box artificially.In the box of half, utilize BIOVUE TM(NJ) transfer pipet pipettes the red blood cell (in normal physiological saline) of 10 μ l 40% in each pillar BioHit for orthoDiagnostic Systems Inc., Raritan.In second half box, the red blood cell as surface antigen listed in the following table 10 (in normal physiological saline) that has of 50 μ l 80% is pipetted in the post of box.At half box that is used for testing, use Ortho BIOVUE TMThe international box of ABE (Ortho Diagnostic Systems Inc., Raritan, NewJersey).At second half box that is used for testing, use (Ortho BIOVUE TMBltk box (Ortho Diagnostic SystemsInc., Raritan, New Jersey).ABE and BHK box all have antibody (shown in following table 10) in each post
Table 10
The ABE box
Post Antibody That is estimated has an A 1The result of rr cell
1 Anti--A +
2 Anti--B -
3 Anti--AB +
4 Anti--D -
5 Anti--CDE -
6 Contrast (thinning agent) -
The RHK box
Post Antibody That is estimated has a R 1R 1The result of K (-) cell
1 Anti--C +
2 Anti--E -
3 Anti--C -
4 Anti--e +
5 Anti--K -
6 Contrast (thinning agent)
When using the ABE box, A 1The rr cell is as sample.When using the RHK box, R 1R1K (-) cell is as sample.Move liquid and be performed until the right from the pillar of leftmost box.Box is at Ortho BIOVUE TMIn the hydro-extractor (Ortho DiagnosticSystems Inc., Raritan NJ) with 794+/-centrifugal 2 minutes of the power of 16xg.And then centrifugal 3 minutes with the power of 1510+/30xg.Assess the post that each has negative reaction (red blood cell is aggegation not), see that whether red blood cell is fully by whole post.If for example, anti--H antibody is transferred to post 2 and just reaction post 2 this moment from the post 1 of ABE box, then false positive reaction can occur.
The results are shown in the table 11, in the post with O shape ring, the positive reaction of all expectations all is positive.Yet the negative reaction of all expectations not all is negative also.Though this reason is not identified, is not the syphonic effect of reagent.Table 11 shows the sum that at every turn is used for the box tested and has the box of false positive reaction and the number and the number percent of post.The insertion method of lining does not influence the result who uses the lining with O shape ring, but remote effect use the result of the lining of no O shape ring, this be because of all false positive reactions all occur in have visible by in the box of the reagent of siphon.
They are 11 years old
Lining insertion method The lining of no O shape ring The lining that O shape ring is arranged
Instrument (vertically) Box 1/96(1％) 1/96(1％)
Post 1/384(0.3％) 1/384(0.3％)
Manually (counter-rotating) Box 9/96(10％) 0/96
Post 10/384(2.6％) 0/384
Amount to Box 10/192(5.2％) 1/192(0.5％)
Post 11/768(1.4％) 1/768(0.1％)

Claims (3)

1. container that is used to carry out CA comprises:
A) one has a last chamber that is used to receive the opening of fluid reaction agent;
B) one is used for the following chamber that receives fluid from last chamber and contain the matrix of separating agglutinator; And
C) will go up the barrier that chamber and following chamber separate for one, this barrier can remain on fluid in the chamber under normal gravity and atmospheric pressure, and can make under greater than the pressure of atmospheric pressure fluid from the chamber flow under the chamber, wherein said barrier comprises a dividing plate.
2. the container of claim 1, wherein said dividing plate have little a center pit that is enough under normal gravity and atmospheric pressure fluid remained in the chamber.
3. the container of claim 2, its median septum is a silicon rubber.
CNB03140653XA 1996-02-02 1997-02-02 Agglutination reaction and separating container Expired - Lifetime CN1252475C (en)

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US08/595,719 US5780248A (en) 1993-07-15 1996-02-02 Foil sealed cassette for agglutination reactions and liner therefor

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US7850917B2 (en) * 2008-03-11 2010-12-14 Ortho-Clinical Diagnostics, Inc. Particle agglutination in a tip
IT1393104B1 (en) * 2009-02-25 2012-04-11 Sentinel Ch S P A TEST TUBE FOR THE COLLECTION, TRANSPORT AND EXTRACTION OF FECI SAMPLES
CN102608336B (en) * 2011-01-19 2014-12-24 刘大基 Disposable transfusion and cross matching experiment combiner
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