CA2117204A1 - Reaction vessel for performing analytical assays - Google Patents
Reaction vessel for performing analytical assaysInfo
- Publication number
- CA2117204A1 CA2117204A1 CA002117204A CA2117204A CA2117204A1 CA 2117204 A1 CA2117204 A1 CA 2117204A1 CA 002117204 A CA002117204 A CA 002117204A CA 2117204 A CA2117204 A CA 2117204A CA 2117204 A1 CA2117204 A1 CA 2117204A1
- Authority
- CA
- Canada
- Prior art keywords
- reaction
- analytical
- reagent
- chambers
- channel
- 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.)
- Abandoned
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
-
- 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
- G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
- G01N33/721—Haemoglobin
- G01N33/723—Glycosylated haemoglobin
-
- 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
- G01N33/72—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood pigments, e.g. haemoglobin, bilirubin or other porphyrins; involving occult blood
- G01N33/721—Haemoglobin
- G01N33/726—Devices
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Hematology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Clinical Laboratory Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
ABSTRACT OF THE DISCLOSURE
Disclosed are a method and device for performing sequential analytical reactions involving a first dry reagent and a second dry reagent comprised of two or more components having different rates of solubilization.
The invention enables one to fully solubilize the components of the second reagent before they are brought into contact with each other to thereby avoid interference with the reaction kinetics which result when one or both of the components are not fully dissolved prior to their being brought into contact.
The invention is especially useful in conjunction with immunoassay formats involving latex bound antibodies and polymeric agglutinators.
Disclosed are a method and device for performing sequential analytical reactions involving a first dry reagent and a second dry reagent comprised of two or more components having different rates of solubilization.
The invention enables one to fully solubilize the components of the second reagent before they are brought into contact with each other to thereby avoid interference with the reaction kinetics which result when one or both of the components are not fully dissolved prior to their being brought into contact.
The invention is especially useful in conjunction with immunoassay formats involving latex bound antibodies and polymeric agglutinators.
Description
2 ~ ~ 72 V ~
REACTION VESSEL FOR
PERFORMING ANALYTICAL ASSAYS
Background of the Inventlon The present invention relates to an analytical assay procedure for determining the amount of an analyte present in a liquid test sample. In pa~ticular, .
the present invention relates to the determination of an analyte in a liquid test sample involving analytical reactions between the analyte and one or more analytical 10 reagents requiring sequential manipulative steps to - ~
make such determination. ~ ~:
. .
In United States Patent 4,990,075 there is dis~
closed a self contained analytical reaction vessel or device and mPthod fbr performing analytical assay - :
:: 15 procedures involving sequential analytical reactions between an analyte in a liquid test sample and one or more analytical reagents which interact with the `~
analyte:to produce a detectable response as a function of the analyte.~ The~ device is described as being particularly useful for performing immunoassaysjwhich : typically require a number of cumbersome manipulative steps such as pipetting, mixing and incubation of the liquid test sample with the analytical reagents.
;~ : MSE-#1836 '' "" '' - '` "'~
~ .
More particularly, this device is described as comprising a closed container having a substantially horizontal axis of rotation and an analytical reagent reaction channel, liquid test sample delivery means for facilitating the flow of a liquid test sample into the reaction channel. The analytical reaction channel comprises one or more reaction zones incorporated with one or more analytical reagents in the dry form. A
first analytical reagent is incorporated into the first 10 reaction zone and a second reagent or reagents into the second reaction zone which is in liquid communication with the first reaction zone. A liquid test sample disposed in the reaction channel can be transported by gravity along the reaction channel between the reaction 15 zones by rotating the device along its horizontal axis of rotation. This device works well when only one reagent is in the second reaction zone since after dissolution of the first analytical reagent the device is simply tilted to bring the re!action fluid into contact wi~h the second dry reagent in the second reaction zone and allowed to remain in this configura~
tion until the second rea~ent completely dissolves.
However, in certain analytical procedures, there must be two or more dry reagents in the second reaction zone. When these reagents have different rates of dissolution, and are being dissolved simultaneously in ~ -the same fluid, certain problems are encountered. For example, if the dissolution rates of the two reagents are slow and the reaction kinetics are fast, most of the reaction will have been compl~ted before all of the reactants are dlssolved thereby interfering with the MSE-#1836 , - ~ ; : . . , : . .
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measurement of the initial kinetics of the reaction, which are the most sensitive part thereof.
, ' ~, In United States patent 4,970,171 there is dis~
closed an analytical method for determining glycated 5 hemoglobin wherein the amounts of both total hemoglobin and glycated hemoglobin derivative are measured and ~
related as a percentage. In this method, a blood ~;
sample is treated with a thiocyanate salt and an oxidant to denature the hemoglobin in the sample 10 thereby converting it to met-hemoglobin. The met-hemoglobin is measured spectrophotometrically to give ~ ~
the total amount of hemoglobin in the sample while the ~-denatured glycated hemoglobin can be measured by immunoassay. This patent describes a par~icle 15 agglutination inhibition assay based on the specific -~ -interaction of an antibody particle reagent and an aggluti~ator. The antibody paxticle reagent comprises ;
the antibody, ox a fragment thereof, bound to a water suspensible particle, e.g. a po].ystyrene or other 20 latex, and the agglu~inator comprises a polymeric material bearing a plurality of epitopic binding sites for the antibody reagent. This sort of immunosassay format is well known ~o those skilled in this art. The above described assay for glycated hemoglobin is well -suited for adaptation to the reaction vessel for perfoxming sequential analytical assays. Thus, placing the dry oxidant/isothiocyanate in the first reaction ~ `
zone'and dissolving it in the reaction fluid containing a blood sample to thereby cause the denaturation of the blood's hemoglobin and rotating the vessel to cause the reaction fluid to come into contact with dried antibody MSE-$1836 ~ ': '. ~
2l~72~
bearing latex and agglutinator as the second reagents in the second reaction zone, facilitates the immuno-agglutination assay upon dissolution of the latex and agglutinator. ~owever, since the latex and agglutinator 5 go into suspension at different rates, a problem can be encount~red since the latex reagent dissolves much more slowly than the agglutinator, the majority of the reaction occurs before all of the reactants are dis-solved. Accordingly, the initial agglutination reac- -~
10 tion, which is the most sensitive part of the assay, cannot be measured with the maximum degree of accuracy attainable for this type of immunoa~say.
It is an object of the pr~sent invention to proYide a me~hod and device which are useful for 15 carrying out the above descrlbed sequential assay while obviating the problem~ associated with the dissolution of two or more analytical reagerlts in the second reaction zone.
The presen~ invention is described in greater 20 detail in the following discussion. Since the latex -~
reagent does not form a true solution with the reaction fluid, the terms dissolution and solution as use~
herein are intended to include the colloidal suspension of the latex particles having antibody bound thereto as 25 well as tru~ solutions.
, I , : . : .: ~ ~:
~ummary of the Inventlon `~
The present invention involves a method for performing sequential analytical reactions for the MSE-#1836 - ~
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determination of an analyte in a test sample comprising the steps of:
(a) providing a closed analytical reagent reaction vessel having a substan~ially horizontal axis of :~
s rotation, which reaction vessel comprises:
i. an analytical reagent reaction channel containing first and second reaction zones in fluid communication with each other which reaction zones are incorporated with a firs~
dry analytical reagent in the first reaction zone and dry compon~nts of a second analytical :~
reagent in the second reaction ~one which zone is divided by one or more septa posi~
tioned therein into at least two integral ~ 8 :~:
solubiliza~ion chambers each of said chambers :
containing a componen~ of the second analyt~
ical reagent, whereby a liquid test sample disposed in said reaction channel can be :~
transported by gravity along the reaction channel between the first and second reaction ;~
æones into and out of the integral solubili~
zation chambers in the second reaction zone by rotating the reaction vessel about its t'~
horizontal axis;
ii. inlet means in fluid communication with the reaction channel for introducing a liquid ; ~est sample into the reaction channel;
~, MSE-#1836 '~ ~
2 ~ ~
(b) introducing ~he liquid test sample into the -~
reaction vessel through the inlet means;
(c) bringing the liquid test sample into contact with the first dry analytical reagent in the first reaction zone to solubilize the first reagent and thereby form a first reaction solution;
(d) rotating the reaction vessel about its axis of rotation in a first direction so that the first reaction solution is transported by gravity away from the first reaction zone along the reaction channel and into contact with the components of :~
the second analytical reagent in the integral reaction chambers to form second, third and optionally additional reaction solutions which are separated from each other by the septum or septa;
(e) maintaining the second, third and optional additional reaction solutions separate from each other for a time sufficient to permit a desired amount of dissolution of the components of the ~
: 20 second analytical reagent in the liguid test ~ :
: sample to take place;
(f) ro~ating the reaction vessel about its axis of rotation in the direction opposite to that of .
the first direction to thereby cause the reaction ::
fluid bearing the components of the second ana~
~ lytical reagent to flow out of the inte~ral :~ chambers, enter the reaction channel and mix with ~:
~: each other so that the second, third and optional ~ :
~, .
MSE-#1836 ~ ~
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-- 7 ~
additional analytical reagents undergo a reaction with each other to provide a detectable response;
and (g) measuring the detectable response.
~- ;. :: : ~ ', .
Also included within the scope of the present ~-~
invention is the reaction vessel designed for carrying .
out the above described method.
~rlef Descriptlon of the Drawlngs Figure 1 represents one embodiment of the prior art reaction vessel. Figure lA depicts the improved reaction vessel of the present invention.
F~gures ~ a~d 2A represent a~o her embodiment of ~he prior ar~ ~nd the improved reaction ve~sel of the present invention.
:~ 15 Figures 3 and 3A-3C represent views of the presently disclosed de~ice through its sidewall during various stages (A-D~ of the method of the present invention.
: De~rip~lon of the Inv~ntlon :
,20 Referring to Figs. 1, lA, 2 and 2A, the device 10 has inner walls 14 which form a delivery chamber 23 : :~
permitting the introduction of a liquid test sample, such as a small amount of blood or other biological fluid to be analyzed, into the device, and, since the delivery chamber is in fluid commullication with reaction MSE - # 18 3 6 : ~ '`";''''~'' . . `.' ' ~, ~` 2:~7~
channel 21, the liquid test sample can enter the reaction channel through the delivery chamber and be caused to flow along the reaction channel by clockwise rotation of the device along its horizontal axis of 5 rotation. ~he test sample is conveniently delivered through capillary dispenser 12 as depicted in Fig. 1. :
Since only a small amount of blood or other biological fluid will typically be introduced through delivery channel 23, additional fluid can be introduced either - ~ ~:
10 through the delivery channel or from another source such as liquid delivery reservoir 26 adapted to contain a buffer and/or liquid reagent for pexforming an : ~
analytical assay procedure. The liquid delivery ~ :
reservoir comprises a reservoir body 27 having a ~ ~
15 depression therein 26 to act as fluid reservoir for ~.
holding the fluid until needed which is covered by a seal or membrane (not shown) which can be removed to:: -allow the fluid in the reservoir 26 to flow into reaction channel 21. Simple manipulation of the device ~ :
20 will cause the liquid test sample optionally carried by the fluid from liquid reservoir 26 to flow into position for viewing through viewing cha~er 42.
In each embodiment, the device has a firs~ reaction :~
zone 29, which is typically disposed in the reaction channel 210 The first reaction zone contains first analytical reagent 28 which is in the dry form and is attached to the innPr wall 16 of said reaction channel or, alternatively, to one of the side walls depicted as -~:
18 a&b in Figs. 3A and 3D. Suitable rotation of the . .
device will bring the reaction fluid in contact with the first dry analytical reagent 28 to facilitate its -:~ :
~ ~.
MSE-#1336 -^ 2 ~ 2 0 ~
g .. ..
dissolution therein. When first dry reagent 28 is adequately dissolved in the reaction fluid carrying the liquid test sample, the device 10 is rotated in the direction opposite to that of the first rotation to ~ ;
5 cause it to be carried by gravity out of the first reaction ~one and into the second reaction zone 34 which is circumscribed by endwall 30, side walls 18 a &
b (depicted in Fig. 3 only) and the outer wall 40 of the reaction channel 21. Disposed in second reaction 10 zone 34 are two separate analytical reagents 32 a & b which, when dissolved in the reaction fluid, take part ~
in a series of chemical or biochemical reactions which ~-facilitate the d~tection and/or quantita~ion of one or more analy~es suspect~d of being present in the biologi-15 cal fluid being analyzed. The reaction chamber of the ~;
devices depicted in Figs. 1 and 2 is a single ~hamber in which dry reagen~s 32 a & b dissolve in the reaction fluid while in contact with each other. As previously ;
mentioned, this can be problemat:ical under certain 20 circumstances. In the device of the present invention, the second reaction zone 34 is clivided into two integral solubilization chambers 20a and 20b by septum 36 which, when the device is rotated so as to cause the reaction : .
liquid to be positioned in ~he ~;econd reaction zone 34, 25 will prevent any intermixing of reayents 32a and 32b while they are dissolving. After the desired degree of dissolution has occurredl the now dissolved reagents 32a and 32b can be mixed by rotating the device back in the opposite direction to cause the separate reagent solutions to flow out of the reaction zone 34 and into reaction channel 21. When reagents 32a and ~2b have reac~ed for a sufficient time to provide the desired MSE-#1836 ,.. ~ ., :,:
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detectable response, the reaction fluid is brought in line with viewing port 42, through which the detectable response is measured. While the drawings and foregoing description depict a singly septum 36 which divides the 5 second reaction zone into two integral reaction chambers, there can be more than one septum included in the second reaction zone, so that the number of septa (n) will divide this zone into n~1 integral reaction chambers.
The device of the present invention is particularly useful in performing an immunoturbidimetric assay for determining the relative amount of a particular~hemo-globin derivative such as the glycated hemoglobin HbAlc in a blood sample. Hemoglobin is a long lived plasma 15 protein that couples non-enzymatically to glucose in the blood. The amount of coupled product (HbAlc) formed increases with increasing blood glucose concen~
tration. Levels of this modified protein are thus indicative of the long term concentration of blood 20 glucose. Such an assay as disclosed in previously mentioned U.S. Patent 4,970,171 involved the steps of~
.
a) treating the blood sample with a thiocyanate salt to denature the hemoglobin present in the ~;~ blood sample in the presence of an oxidant ~o convert the denatured hemoglobin to its met form, b) assaying~the denatured blood sample to determine the total amount of met-hemoglobin present therein; ~
`, :.'"''.:':
MSE-#1836 '""' .;, 21 ~2~ :
. , c) assaying the denatured blood sample by immunoassay for the amount of hemoglobin Alc derivative present therein;
d) calculating the relative amount of hemoglobin ~ ;
that is in the form of glycated hemoglobin Alc compared to the total amount of hemoglobin present in the blood sample.
The test device of previously mentioned U.S.
Patent 4r990~075 is well suited for carrying out this ; ; `~
lO sort of assay since the blood sample can be mixed with the thiocyanate/oxidizer in the first reactionlzone and, when dissolution is complete, the resulting solution can be analyzed for total denatured met hemo-globin and then transported to the second reaction zone - ~5 where it is contacted with the reagents necessary for carrying out the immuno assay. This method works well when the immunoassay is of the ELISA type. However, ;~
when the immunoassay is of the latex bound antibody agglutination type where an antibody, or fragment 20 thereof, specific for hemoglobin Alc is bound to a water suspensible particle (e.g. polystyrene or other ~ -latex) and the agglutinator contains a plurality of epitopic binding sites for the antibody, the previously described difficulties are encountered due to the -~
~ 25 varying of the dissolution rates of the latex bound ; antibody and the agglutinator which are predisposed in the second reaction zone. ~he problems associated with `;
the differential dissolution rates is eliminated by the present invention. Figs. 3 and 3a-3c represent a view of second reaction zone 34 of the present invention MSE-#1836 2~172~
,.
taken through its bottom wall 40. The following description of the method and device of the present invention involves the determination of hemoglobin Alc in a blood sample using the latex bound antibody/
agglutinator immunoassay technique. Referring to Fig.
la, a drop of blood to be analyzed is intr~duced into the device 10 via capillary 12 through delivery channel 23 and the reagent fluid comprising lithium thiocyanate in a glycine buffer solution is introduced by removing 10 the covering layer (not shown) from the fluid reservoir 26 to provide a solution of the blood sample in the reaction fluid. The fluid is brought in contact with the first dry reagent 28 which is a ferricyanide salt as oxidant which in combination with the thiocyanate 15 causes denaturation of the hemoglobin and its conver-sion to denatured met-hemoglobin. After sufficient time for dissolution of the ferricyanate/thiocyanate ~-with the resulting denaturation of the hemoqlobln has passed (typically about two to five minutes) the device ~ ~ -is rotated l/8 turn clockwise to cause the reaction fluid to cover viewing port 42. At this point a measurement of total met-hemoglobin is taken such as by measuring the reaction fluid's absorbance at a wavelength of 540 nm. After gaking this measurement, the device is rotated another 3/8 turn clockwise to cause the reaction fluid to flow into the second reaction zone 34 `~
which is divided into two solubilization chambers by septum 36 located in the middle of the reaction zone.
Referring to Fig. 3, there is depicted a septum 36 and the reagent components 32a (the latex bound antibody) and 32b Ithe agglutinator) deposited in the two separate solubilization chambers 20 a & b of the second reaction zone 34.
MSE-#1836 `
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In Fig. 3B there is represented the reaction fluid ~ :
being delivered toward the second reaction zone 34 but before reaching the septum 36. Figure 3C represents the liquid reagent having flowed past the septum 36 and : .
5 into the solubilization chambers 20a and 20b. One chamber is enclosed by the walls 36, 18a and 30 and the ~ :
second chamber i~ enclosed by the walls 36, 18b and 30.
One portion of the reaction fluid dissolves reagent component 32a to give a solution 42a while the other 10 portion of the liquid dissolves reagent component 32b to give solution 42b. Since the solubilization of the two reagent components occurs separately in the ~ -integral chambers, no mixing of the two compone~ts takes place at this stage. Figure 3C represents the 15 device having been rotated 3/8 turn in a counter :
clockwise direction thereby causing the liquid reagents 42a and 42b to flow out of the ~olubilizatlon chambers : ~:
into the reactio~ channel 21 where they mix to form a homogenous reaction mixture. In the case of the agglutination immunoassay for hemoglobin Alc under con~idera~ion, the homogenous reaction mixture will undergo a reaction with the HbAlc analyte in which the :
analyte will compete for the epitope sites on the : agglutinator wlth the HbAlc antibody bound to the :
latex~ Since the analyte concentration and degrae of agglutination will be in an inverse relationship, the : higher the concentration of analyte the less agglutination will occur. By measuring the degree of ~:
agglutination, such as by well known naphthometric or light density techniques, an accurate measurement of the analyte, hemoglobin Alc in the present case, can be made.
MSE-#1836
REACTION VESSEL FOR
PERFORMING ANALYTICAL ASSAYS
Background of the Inventlon The present invention relates to an analytical assay procedure for determining the amount of an analyte present in a liquid test sample. In pa~ticular, .
the present invention relates to the determination of an analyte in a liquid test sample involving analytical reactions between the analyte and one or more analytical 10 reagents requiring sequential manipulative steps to - ~
make such determination. ~ ~:
. .
In United States Patent 4,990,075 there is dis~
closed a self contained analytical reaction vessel or device and mPthod fbr performing analytical assay - :
:: 15 procedures involving sequential analytical reactions between an analyte in a liquid test sample and one or more analytical reagents which interact with the `~
analyte:to produce a detectable response as a function of the analyte.~ The~ device is described as being particularly useful for performing immunoassaysjwhich : typically require a number of cumbersome manipulative steps such as pipetting, mixing and incubation of the liquid test sample with the analytical reagents.
;~ : MSE-#1836 '' "" '' - '` "'~
~ .
More particularly, this device is described as comprising a closed container having a substantially horizontal axis of rotation and an analytical reagent reaction channel, liquid test sample delivery means for facilitating the flow of a liquid test sample into the reaction channel. The analytical reaction channel comprises one or more reaction zones incorporated with one or more analytical reagents in the dry form. A
first analytical reagent is incorporated into the first 10 reaction zone and a second reagent or reagents into the second reaction zone which is in liquid communication with the first reaction zone. A liquid test sample disposed in the reaction channel can be transported by gravity along the reaction channel between the reaction 15 zones by rotating the device along its horizontal axis of rotation. This device works well when only one reagent is in the second reaction zone since after dissolution of the first analytical reagent the device is simply tilted to bring the re!action fluid into contact wi~h the second dry reagent in the second reaction zone and allowed to remain in this configura~
tion until the second rea~ent completely dissolves.
However, in certain analytical procedures, there must be two or more dry reagents in the second reaction zone. When these reagents have different rates of dissolution, and are being dissolved simultaneously in ~ -the same fluid, certain problems are encountered. For example, if the dissolution rates of the two reagents are slow and the reaction kinetics are fast, most of the reaction will have been compl~ted before all of the reactants are dlssolved thereby interfering with the MSE-#1836 , - ~ ; : . . , : . .
2.~ 1 72 0 ~
~ . .
.. : .
measurement of the initial kinetics of the reaction, which are the most sensitive part thereof.
, ' ~, In United States patent 4,970,171 there is dis~
closed an analytical method for determining glycated 5 hemoglobin wherein the amounts of both total hemoglobin and glycated hemoglobin derivative are measured and ~
related as a percentage. In this method, a blood ~;
sample is treated with a thiocyanate salt and an oxidant to denature the hemoglobin in the sample 10 thereby converting it to met-hemoglobin. The met-hemoglobin is measured spectrophotometrically to give ~ ~
the total amount of hemoglobin in the sample while the ~-denatured glycated hemoglobin can be measured by immunoassay. This patent describes a par~icle 15 agglutination inhibition assay based on the specific -~ -interaction of an antibody particle reagent and an aggluti~ator. The antibody paxticle reagent comprises ;
the antibody, ox a fragment thereof, bound to a water suspensible particle, e.g. a po].ystyrene or other 20 latex, and the agglu~inator comprises a polymeric material bearing a plurality of epitopic binding sites for the antibody reagent. This sort of immunosassay format is well known ~o those skilled in this art. The above described assay for glycated hemoglobin is well -suited for adaptation to the reaction vessel for perfoxming sequential analytical assays. Thus, placing the dry oxidant/isothiocyanate in the first reaction ~ `
zone'and dissolving it in the reaction fluid containing a blood sample to thereby cause the denaturation of the blood's hemoglobin and rotating the vessel to cause the reaction fluid to come into contact with dried antibody MSE-$1836 ~ ': '. ~
2l~72~
bearing latex and agglutinator as the second reagents in the second reaction zone, facilitates the immuno-agglutination assay upon dissolution of the latex and agglutinator. ~owever, since the latex and agglutinator 5 go into suspension at different rates, a problem can be encount~red since the latex reagent dissolves much more slowly than the agglutinator, the majority of the reaction occurs before all of the reactants are dis-solved. Accordingly, the initial agglutination reac- -~
10 tion, which is the most sensitive part of the assay, cannot be measured with the maximum degree of accuracy attainable for this type of immunoa~say.
It is an object of the pr~sent invention to proYide a me~hod and device which are useful for 15 carrying out the above descrlbed sequential assay while obviating the problem~ associated with the dissolution of two or more analytical reagerlts in the second reaction zone.
The presen~ invention is described in greater 20 detail in the following discussion. Since the latex -~
reagent does not form a true solution with the reaction fluid, the terms dissolution and solution as use~
herein are intended to include the colloidal suspension of the latex particles having antibody bound thereto as 25 well as tru~ solutions.
, I , : . : .: ~ ~:
~ummary of the Inventlon `~
The present invention involves a method for performing sequential analytical reactions for the MSE-#1836 - ~
,,~ .' '.'... ,.',.
",~, 2~172~
determination of an analyte in a test sample comprising the steps of:
(a) providing a closed analytical reagent reaction vessel having a substan~ially horizontal axis of :~
s rotation, which reaction vessel comprises:
i. an analytical reagent reaction channel containing first and second reaction zones in fluid communication with each other which reaction zones are incorporated with a firs~
dry analytical reagent in the first reaction zone and dry compon~nts of a second analytical :~
reagent in the second reaction ~one which zone is divided by one or more septa posi~
tioned therein into at least two integral ~ 8 :~:
solubiliza~ion chambers each of said chambers :
containing a componen~ of the second analyt~
ical reagent, whereby a liquid test sample disposed in said reaction channel can be :~
transported by gravity along the reaction channel between the first and second reaction ;~
æones into and out of the integral solubili~
zation chambers in the second reaction zone by rotating the reaction vessel about its t'~
horizontal axis;
ii. inlet means in fluid communication with the reaction channel for introducing a liquid ; ~est sample into the reaction channel;
~, MSE-#1836 '~ ~
2 ~ ~
(b) introducing ~he liquid test sample into the -~
reaction vessel through the inlet means;
(c) bringing the liquid test sample into contact with the first dry analytical reagent in the first reaction zone to solubilize the first reagent and thereby form a first reaction solution;
(d) rotating the reaction vessel about its axis of rotation in a first direction so that the first reaction solution is transported by gravity away from the first reaction zone along the reaction channel and into contact with the components of :~
the second analytical reagent in the integral reaction chambers to form second, third and optionally additional reaction solutions which are separated from each other by the septum or septa;
(e) maintaining the second, third and optional additional reaction solutions separate from each other for a time sufficient to permit a desired amount of dissolution of the components of the ~
: 20 second analytical reagent in the liguid test ~ :
: sample to take place;
(f) ro~ating the reaction vessel about its axis of rotation in the direction opposite to that of .
the first direction to thereby cause the reaction ::
fluid bearing the components of the second ana~
~ lytical reagent to flow out of the inte~ral :~ chambers, enter the reaction channel and mix with ~:
~: each other so that the second, third and optional ~ :
~, .
MSE-#1836 ~ ~
': ' ', 2 ~
-- 7 ~
additional analytical reagents undergo a reaction with each other to provide a detectable response;
and (g) measuring the detectable response.
~- ;. :: : ~ ', .
Also included within the scope of the present ~-~
invention is the reaction vessel designed for carrying .
out the above described method.
~rlef Descriptlon of the Drawlngs Figure 1 represents one embodiment of the prior art reaction vessel. Figure lA depicts the improved reaction vessel of the present invention.
F~gures ~ a~d 2A represent a~o her embodiment of ~he prior ar~ ~nd the improved reaction ve~sel of the present invention.
:~ 15 Figures 3 and 3A-3C represent views of the presently disclosed de~ice through its sidewall during various stages (A-D~ of the method of the present invention.
: De~rip~lon of the Inv~ntlon :
,20 Referring to Figs. 1, lA, 2 and 2A, the device 10 has inner walls 14 which form a delivery chamber 23 : :~
permitting the introduction of a liquid test sample, such as a small amount of blood or other biological fluid to be analyzed, into the device, and, since the delivery chamber is in fluid commullication with reaction MSE - # 18 3 6 : ~ '`";''''~'' . . `.' ' ~, ~` 2:~7~
channel 21, the liquid test sample can enter the reaction channel through the delivery chamber and be caused to flow along the reaction channel by clockwise rotation of the device along its horizontal axis of 5 rotation. ~he test sample is conveniently delivered through capillary dispenser 12 as depicted in Fig. 1. :
Since only a small amount of blood or other biological fluid will typically be introduced through delivery channel 23, additional fluid can be introduced either - ~ ~:
10 through the delivery channel or from another source such as liquid delivery reservoir 26 adapted to contain a buffer and/or liquid reagent for pexforming an : ~
analytical assay procedure. The liquid delivery ~ :
reservoir comprises a reservoir body 27 having a ~ ~
15 depression therein 26 to act as fluid reservoir for ~.
holding the fluid until needed which is covered by a seal or membrane (not shown) which can be removed to:: -allow the fluid in the reservoir 26 to flow into reaction channel 21. Simple manipulation of the device ~ :
20 will cause the liquid test sample optionally carried by the fluid from liquid reservoir 26 to flow into position for viewing through viewing cha~er 42.
In each embodiment, the device has a firs~ reaction :~
zone 29, which is typically disposed in the reaction channel 210 The first reaction zone contains first analytical reagent 28 which is in the dry form and is attached to the innPr wall 16 of said reaction channel or, alternatively, to one of the side walls depicted as -~:
18 a&b in Figs. 3A and 3D. Suitable rotation of the . .
device will bring the reaction fluid in contact with the first dry analytical reagent 28 to facilitate its -:~ :
~ ~.
MSE-#1336 -^ 2 ~ 2 0 ~
g .. ..
dissolution therein. When first dry reagent 28 is adequately dissolved in the reaction fluid carrying the liquid test sample, the device 10 is rotated in the direction opposite to that of the first rotation to ~ ;
5 cause it to be carried by gravity out of the first reaction ~one and into the second reaction zone 34 which is circumscribed by endwall 30, side walls 18 a &
b (depicted in Fig. 3 only) and the outer wall 40 of the reaction channel 21. Disposed in second reaction 10 zone 34 are two separate analytical reagents 32 a & b which, when dissolved in the reaction fluid, take part ~
in a series of chemical or biochemical reactions which ~-facilitate the d~tection and/or quantita~ion of one or more analy~es suspect~d of being present in the biologi-15 cal fluid being analyzed. The reaction chamber of the ~;
devices depicted in Figs. 1 and 2 is a single ~hamber in which dry reagen~s 32 a & b dissolve in the reaction fluid while in contact with each other. As previously ;
mentioned, this can be problemat:ical under certain 20 circumstances. In the device of the present invention, the second reaction zone 34 is clivided into two integral solubilization chambers 20a and 20b by septum 36 which, when the device is rotated so as to cause the reaction : .
liquid to be positioned in ~he ~;econd reaction zone 34, 25 will prevent any intermixing of reayents 32a and 32b while they are dissolving. After the desired degree of dissolution has occurredl the now dissolved reagents 32a and 32b can be mixed by rotating the device back in the opposite direction to cause the separate reagent solutions to flow out of the reaction zone 34 and into reaction channel 21. When reagents 32a and ~2b have reac~ed for a sufficient time to provide the desired MSE-#1836 ,.. ~ ., :,:
~' ~. ' ''''',~' 2 ~ 1 7 ~
detectable response, the reaction fluid is brought in line with viewing port 42, through which the detectable response is measured. While the drawings and foregoing description depict a singly septum 36 which divides the 5 second reaction zone into two integral reaction chambers, there can be more than one septum included in the second reaction zone, so that the number of septa (n) will divide this zone into n~1 integral reaction chambers.
The device of the present invention is particularly useful in performing an immunoturbidimetric assay for determining the relative amount of a particular~hemo-globin derivative such as the glycated hemoglobin HbAlc in a blood sample. Hemoglobin is a long lived plasma 15 protein that couples non-enzymatically to glucose in the blood. The amount of coupled product (HbAlc) formed increases with increasing blood glucose concen~
tration. Levels of this modified protein are thus indicative of the long term concentration of blood 20 glucose. Such an assay as disclosed in previously mentioned U.S. Patent 4,970,171 involved the steps of~
.
a) treating the blood sample with a thiocyanate salt to denature the hemoglobin present in the ~;~ blood sample in the presence of an oxidant ~o convert the denatured hemoglobin to its met form, b) assaying~the denatured blood sample to determine the total amount of met-hemoglobin present therein; ~
`, :.'"''.:':
MSE-#1836 '""' .;, 21 ~2~ :
. , c) assaying the denatured blood sample by immunoassay for the amount of hemoglobin Alc derivative present therein;
d) calculating the relative amount of hemoglobin ~ ;
that is in the form of glycated hemoglobin Alc compared to the total amount of hemoglobin present in the blood sample.
The test device of previously mentioned U.S.
Patent 4r990~075 is well suited for carrying out this ; ; `~
lO sort of assay since the blood sample can be mixed with the thiocyanate/oxidizer in the first reactionlzone and, when dissolution is complete, the resulting solution can be analyzed for total denatured met hemo-globin and then transported to the second reaction zone - ~5 where it is contacted with the reagents necessary for carrying out the immuno assay. This method works well when the immunoassay is of the ELISA type. However, ;~
when the immunoassay is of the latex bound antibody agglutination type where an antibody, or fragment 20 thereof, specific for hemoglobin Alc is bound to a water suspensible particle (e.g. polystyrene or other ~ -latex) and the agglutinator contains a plurality of epitopic binding sites for the antibody, the previously described difficulties are encountered due to the -~
~ 25 varying of the dissolution rates of the latex bound ; antibody and the agglutinator which are predisposed in the second reaction zone. ~he problems associated with `;
the differential dissolution rates is eliminated by the present invention. Figs. 3 and 3a-3c represent a view of second reaction zone 34 of the present invention MSE-#1836 2~172~
,.
taken through its bottom wall 40. The following description of the method and device of the present invention involves the determination of hemoglobin Alc in a blood sample using the latex bound antibody/
agglutinator immunoassay technique. Referring to Fig.
la, a drop of blood to be analyzed is intr~duced into the device 10 via capillary 12 through delivery channel 23 and the reagent fluid comprising lithium thiocyanate in a glycine buffer solution is introduced by removing 10 the covering layer (not shown) from the fluid reservoir 26 to provide a solution of the blood sample in the reaction fluid. The fluid is brought in contact with the first dry reagent 28 which is a ferricyanide salt as oxidant which in combination with the thiocyanate 15 causes denaturation of the hemoglobin and its conver-sion to denatured met-hemoglobin. After sufficient time for dissolution of the ferricyanate/thiocyanate ~-with the resulting denaturation of the hemoqlobln has passed (typically about two to five minutes) the device ~ ~ -is rotated l/8 turn clockwise to cause the reaction fluid to cover viewing port 42. At this point a measurement of total met-hemoglobin is taken such as by measuring the reaction fluid's absorbance at a wavelength of 540 nm. After gaking this measurement, the device is rotated another 3/8 turn clockwise to cause the reaction fluid to flow into the second reaction zone 34 `~
which is divided into two solubilization chambers by septum 36 located in the middle of the reaction zone.
Referring to Fig. 3, there is depicted a septum 36 and the reagent components 32a (the latex bound antibody) and 32b Ithe agglutinator) deposited in the two separate solubilization chambers 20 a & b of the second reaction zone 34.
MSE-#1836 `
: . '" ''~ ,, ~
'' ~ ;'''~;' 2~.~72~
In Fig. 3B there is represented the reaction fluid ~ :
being delivered toward the second reaction zone 34 but before reaching the septum 36. Figure 3C represents the liquid reagent having flowed past the septum 36 and : .
5 into the solubilization chambers 20a and 20b. One chamber is enclosed by the walls 36, 18a and 30 and the ~ :
second chamber i~ enclosed by the walls 36, 18b and 30.
One portion of the reaction fluid dissolves reagent component 32a to give a solution 42a while the other 10 portion of the liquid dissolves reagent component 32b to give solution 42b. Since the solubilization of the two reagent components occurs separately in the ~ -integral chambers, no mixing of the two compone~ts takes place at this stage. Figure 3C represents the 15 device having been rotated 3/8 turn in a counter :
clockwise direction thereby causing the liquid reagents 42a and 42b to flow out of the ~olubilizatlon chambers : ~:
into the reactio~ channel 21 where they mix to form a homogenous reaction mixture. In the case of the agglutination immunoassay for hemoglobin Alc under con~idera~ion, the homogenous reaction mixture will undergo a reaction with the HbAlc analyte in which the :
analyte will compete for the epitope sites on the : agglutinator wlth the HbAlc antibody bound to the :
latex~ Since the analyte concentration and degrae of agglutination will be in an inverse relationship, the : higher the concentration of analyte the less agglutination will occur. By measuring the degree of ~:
agglutination, such as by well known naphthometric or light density techniques, an accurate measurement of the analyte, hemoglobin Alc in the present case, can be made.
MSE-#1836
Claims (10)
1. A method for performing sequential analytical reactions to determine an analyte in a test sample, which method comprises the steps of:
(a) providing a closed analytical reagent reaction vessel having a substantially horizontal axis of rotation and comprising:
i. an analytical reagent reaction channel containing first and second reaction zones in fluid communication with each other which reaction zones are incorporated with a first dry analytical reagent in the first reaction zone and dry components of a second analytical reagent in the second reaction zone which zone is divided by one or more septa posi-tioned therein into at least two integral solubilization chambers each of said chambers containing a component of the second analyt-ical reagent whereby a liquid test sample disposed in said reaction channel can be transported by gravity along the reaction channel between the first and second reaction zones into and out of the integral solubili-zation chambers in the second reaction zone by rotating the reaction vessel about its horizontal axis;
ii. inlet means in fluid communication with the reaction channel for introducing a liquid test sample into the reaction channel;
(b) introducing the liquid test sample into the reaction vessel through the inlet means;
(c) bringing the liquid test sample into contact with the first analytical reagent in the first reaction zone to solubilize the first reagent and thereby form a first reaction mixture;
(d) rotating the reaction vessel about its axis of rotation in a first direction so that the first reaction mixture is transported by gravity away from the first reaction zone along the reaction channel and into contact with the components of the second analytical reagent in the integral reaction chambers to form second and third reaction mixtures which are separated from each other by the septum or septa;
(e) maintaining the second and third reaction mixtures separate from each other for a time sufficient to permit a desired amount of dissolu-tion of the components of the second analytical reagent in the liquid test sample to take place;
(f) rotating the reaction vessel about its axis of rotation in the direction opposite to that of the first direction to thereby cause the reaction fluid bearing the components of the second analytical reagent to leave the integral chambers, enter the reaction channel and mix with each other so that the first and second analytical reagents undergo a reaction with each other to provide a detectable response; and (g) measuring the detectable response.
(a) providing a closed analytical reagent reaction vessel having a substantially horizontal axis of rotation and comprising:
i. an analytical reagent reaction channel containing first and second reaction zones in fluid communication with each other which reaction zones are incorporated with a first dry analytical reagent in the first reaction zone and dry components of a second analytical reagent in the second reaction zone which zone is divided by one or more septa posi-tioned therein into at least two integral solubilization chambers each of said chambers containing a component of the second analyt-ical reagent whereby a liquid test sample disposed in said reaction channel can be transported by gravity along the reaction channel between the first and second reaction zones into and out of the integral solubili-zation chambers in the second reaction zone by rotating the reaction vessel about its horizontal axis;
ii. inlet means in fluid communication with the reaction channel for introducing a liquid test sample into the reaction channel;
(b) introducing the liquid test sample into the reaction vessel through the inlet means;
(c) bringing the liquid test sample into contact with the first analytical reagent in the first reaction zone to solubilize the first reagent and thereby form a first reaction mixture;
(d) rotating the reaction vessel about its axis of rotation in a first direction so that the first reaction mixture is transported by gravity away from the first reaction zone along the reaction channel and into contact with the components of the second analytical reagent in the integral reaction chambers to form second and third reaction mixtures which are separated from each other by the septum or septa;
(e) maintaining the second and third reaction mixtures separate from each other for a time sufficient to permit a desired amount of dissolu-tion of the components of the second analytical reagent in the liquid test sample to take place;
(f) rotating the reaction vessel about its axis of rotation in the direction opposite to that of the first direction to thereby cause the reaction fluid bearing the components of the second analytical reagent to leave the integral chambers, enter the reaction channel and mix with each other so that the first and second analytical reagents undergo a reaction with each other to provide a detectable response; and (g) measuring the detectable response.
2. The method of Claim 1 wherein the reaction vessel has a reaction viewing zone in the form of an area having transparent walls in fluid communication with the first and second reaction zones and the detectable response is measured by taking readings through the transparent walls.
3. The method of Claim 1 wherein there is a single septum in the second reaction zone dividing it into two integral chambers.
4. The method of Claim 1 wherein the analyte is hemoglobin Alc and the first analytical reagent is capable of denaturing hemoglobin.
5. The method of Claim 4 wherein the first analytical reagent is an oxidant/isothiocyanate combination.
6. The method of Claim 1 wherein the components of the second analytical reagent are a latex bound antibody and an agglutinator comprising a polymeric material bearing a plurality of epitopic binding sites for the antibody.
7. The method of Claim 6 wherein the antibody is specific for hemoglobin Alc.
8. A method for performing a sequential analytical reaction to determine the amount of hemoglobin Alc in a blood sample, which method comprises the steps of:
(a) providing a closed analytical reaction vessel having a substantially horizontal axis of rotation and comprising:
i. an analytical reagent reaction channel containing first and second reaction zones in fluid communication with each other which reaction zones are incorporated with a dry oxidant/isothiocyanate capable of denaturing hemoglobin in the first reaction zone and dry latex bound antibody specific for HbAlc/polymeric agglutinator containing a plurality of epitopic binding sites for the HbAlc antibody in the second reaction zone which is divided into two integral solubili-zation chambers by a septum in its midsection with one chamber containing the dry latex bound antibody and the other containing the dry agglutinator, whereby the blood sample disposed in the reaction channel can be transported by gravity along the reaction channel between the first and second reaction zones into and out of the integral solubiliza-tion chambers by rotating the reaction vessel about its horizontal axis;
ii. inlet means in fluid communication with the reaction vessel for introducing the blood sample into the reaction channel;
(b) introducing the blood sample into the reaction vessel through the inlet means;
(c) bringing the blood sample into contact with the oxidant isothiocyanate in the first reaction zone to solubilize it thereby forming a first reaction mixture containing denatured hemoglobin, and determining the amount of hemoglobin in the blood sample;
(d) rotating the reaction vessel about its axis of rotation in a first direction so that the first reaction mixture is transported by gravity away from the first reaction zone along the reaction channel and into contact with the latex bound antibody and polymeric agglutinator while keeping them separated by the septum;
(e) maintaining the separated latex bound antibody and agglutinator in contact with the reaction fluid for a time sufficient to permit the desired amount of dissolution to take place;
(f) rotating the reaction vessel about its axis of rotation in the direction opposite to that of the first direction to thereby cause the reaction fluid bearing the latex bound antibody and polymeric agglutinator to leave the integral solubilization chambers, enter the reaction channel and mix with each other so that the latex bound antibody and agglutinator enter into a competitive immunoreac-tion with each other and the hemoglobin Alc in the blood sample; and (g) measuring the amount of agglutination which takes place to thereby determine the amount of hemoglobin Alc in the blood sample.
(a) providing a closed analytical reaction vessel having a substantially horizontal axis of rotation and comprising:
i. an analytical reagent reaction channel containing first and second reaction zones in fluid communication with each other which reaction zones are incorporated with a dry oxidant/isothiocyanate capable of denaturing hemoglobin in the first reaction zone and dry latex bound antibody specific for HbAlc/polymeric agglutinator containing a plurality of epitopic binding sites for the HbAlc antibody in the second reaction zone which is divided into two integral solubili-zation chambers by a septum in its midsection with one chamber containing the dry latex bound antibody and the other containing the dry agglutinator, whereby the blood sample disposed in the reaction channel can be transported by gravity along the reaction channel between the first and second reaction zones into and out of the integral solubiliza-tion chambers by rotating the reaction vessel about its horizontal axis;
ii. inlet means in fluid communication with the reaction vessel for introducing the blood sample into the reaction channel;
(b) introducing the blood sample into the reaction vessel through the inlet means;
(c) bringing the blood sample into contact with the oxidant isothiocyanate in the first reaction zone to solubilize it thereby forming a first reaction mixture containing denatured hemoglobin, and determining the amount of hemoglobin in the blood sample;
(d) rotating the reaction vessel about its axis of rotation in a first direction so that the first reaction mixture is transported by gravity away from the first reaction zone along the reaction channel and into contact with the latex bound antibody and polymeric agglutinator while keeping them separated by the septum;
(e) maintaining the separated latex bound antibody and agglutinator in contact with the reaction fluid for a time sufficient to permit the desired amount of dissolution to take place;
(f) rotating the reaction vessel about its axis of rotation in the direction opposite to that of the first direction to thereby cause the reaction fluid bearing the latex bound antibody and polymeric agglutinator to leave the integral solubilization chambers, enter the reaction channel and mix with each other so that the latex bound antibody and agglutinator enter into a competitive immunoreac-tion with each other and the hemoglobin Alc in the blood sample; and (g) measuring the amount of agglutination which takes place to thereby determine the amount of hemoglobin Alc in the blood sample.
9. The method of Claim 8 wherein the percent of glycated hemoglobin in the blood sample is determined by dividing the concentration of HbAlc by the concentra-tion of total hemoglobin.
10. In combination with a reaction vessel for per-forming analytical assays comprising a closed container having a substantially horizontal axis of rotation said vessel containing an analytical reagent reaction channel having first and second reaction zones in fluid communication with each other and incorporated with first and second analytical reagents which interact with an analyte in a liquid test sample disposed in the reaction channel to provide a detectable response the improvement which comprises a septum or septa in the second reaction zone positioned therein to divide it into two or more integral solubilization chambers, with each of said chambers containing a dry component of the second analytical reagent whereby the liquid test sample disposed in the reaction channel can be trans-ported by gravity between the first and second reaction zones and into and out of the integral solubilization chambers by rotating the reaction vessel about its axis of rotation and the dry components of the second analytical reagent can be separately solubilized in the liquid test sample to form reagent component solutions which can be combined in the reagent reaction channel to form the second analytical reagent.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US08/032,643 | 1993-03-17 | ||
| US08/032,643 US5372948A (en) | 1993-03-17 | 1993-03-17 | Assay and reaction vessel with a compartmentalized solubilization chamber |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2117204A1 true CA2117204A1 (en) | 1994-09-18 |
Family
ID=21866035
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002117204A Abandoned CA2117204A1 (en) | 1993-03-17 | 1994-03-08 | Reaction vessel for performing analytical assays |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US5372948A (en) |
| EP (1) | EP0616216A3 (en) |
| JP (1) | JP3447360B2 (en) |
| AU (1) | AU669603B2 (en) |
| CA (1) | CA2117204A1 (en) |
Families Citing this family (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR970706902A (en) * | 1995-09-12 | 1997-12-01 | 로드릭 리차드 제이 | DEVICE AND METHOD FOR DNA AMPLIFICATION AND ASSAY |
| EP0848979A4 (en) * | 1996-05-31 | 1999-01-13 | Toray Industries | Filter cloth and filter |
| EP1752755B1 (en) * | 2005-08-10 | 2015-05-06 | Roche Diagnostics GmbH | sampling and dosing device with an integrate container for fluid |
| WO2009044552A1 (en) | 2007-10-04 | 2009-04-09 | Panasonic Corporation | Analysis device, and analysis apparatus and method using the same |
| JP5288768B2 (en) * | 2007-10-18 | 2013-09-11 | パナソニック株式会社 | Analysis container and analyzer |
| KR100798471B1 (en) * | 2007-10-08 | 2008-01-28 | 주식회사 인포피아 | Reaction cassette for measuring glycated hemoglobin and measuring method thereof |
| KR100799354B1 (en) * | 2007-11-08 | 2008-01-30 | 주식회사 인포피아 | Reagent vessel |
| WO2009112038A2 (en) * | 2008-03-14 | 2009-09-17 | Scandinavian Micro Biodevices Aps | A microfluidic system and a method of performing a test |
| US9182353B2 (en) | 2010-07-22 | 2015-11-10 | Hach Company | Lab-on-a-chip for alkalinity analysis |
| TWI476406B (en) * | 2011-11-10 | 2015-03-11 | Apex Biotechnology Corp | Reaction cassette and assay device |
| US9180449B2 (en) | 2012-06-12 | 2015-11-10 | Hach Company | Mobile water analysis |
| USD768872S1 (en) | 2012-12-12 | 2016-10-11 | Hach Company | Cuvette for a water analysis instrument |
| CN105021544B (en) | 2015-07-03 | 2018-02-02 | 艾康生物技术(杭州)有限公司 | Detecting system |
| CN105013546B (en) | 2015-07-03 | 2017-12-08 | 艾康生物技术(杭州)有限公司 | Biological sample reaction box |
| US10775370B2 (en) * | 2015-07-17 | 2020-09-15 | Stat-Diagnostica & Innovation, S.L. | Fluidic system for performing assays |
| MX2019000592A (en) * | 2016-07-18 | 2019-09-27 | Siemens Healthcare Diagnostics Inc | Liquid analytical reagent dispensing apparatus and analytical kits and methods of use related thereto. |
| EP3521830A4 (en) * | 2016-09-29 | 2020-06-17 | Green Cross Medical Science | Separable cassette for measuring glycated hemoglobin |
| EP3537156A4 (en) | 2016-11-04 | 2020-07-15 | Green Cross Medical Science | Measurement method for glycated hemoglobin ratio |
| WO2020180552A1 (en) * | 2019-03-04 | 2020-09-10 | Siemens Healthcare Diagnostics Inc. | Dye-based liquid reagent volume indicator for use in analyte detection assays |
| CN110596409B (en) * | 2019-08-16 | 2020-12-29 | 深圳市朴瑞生物科技有限公司 | Reagent pack applied to rapid instant detection and detection equipment thereof |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4918025A (en) * | 1987-03-03 | 1990-04-17 | Pb Diagnostic Systems, Inc. | Self contained immunoassay element |
| US4847209A (en) * | 1987-11-09 | 1989-07-11 | Miles Inc. | Latex agglutination immunoassay in the presence of hemoglobin |
| US4970171A (en) * | 1987-11-09 | 1990-11-13 | Miles Inc. | Denaturant reagents for convenient determination of hemoglobin derivatives in blood |
| US5162237A (en) * | 1988-04-11 | 1992-11-10 | Miles Inc. | Reaction cassette for preforming sequential analytical assays by noncentrifugal and noncapillary manipulations |
| US4990075A (en) * | 1988-04-11 | 1991-02-05 | Miles Inc. | Reaction vessel for performing sequential analytical assays |
| US5084397A (en) * | 1988-04-11 | 1992-01-28 | Miles Inc. | Method and apparatus for controlled reagent deposition in reaction cassettes and the like |
| US5104813A (en) * | 1989-04-13 | 1992-04-14 | Biotrack, Inc. | Dilution and mixing cartridge |
| IL94408A0 (en) * | 1989-07-11 | 1991-03-10 | Miles Inc | Method,reaction cassette and kit for performing analytical assays |
| US5120503A (en) * | 1989-07-14 | 1992-06-09 | Eastman Kodak Company | Extracting device for extracting antigens |
-
1993
- 1993-03-17 US US08/032,643 patent/US5372948A/en not_active Expired - Fee Related
-
1994
- 1994-03-08 CA CA002117204A patent/CA2117204A1/en not_active Abandoned
- 1994-03-09 EP EP94103595A patent/EP0616216A3/en not_active Withdrawn
- 1994-03-11 AU AU57762/94A patent/AU669603B2/en not_active Ceased
- 1994-03-16 JP JP04587194A patent/JP3447360B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| EP0616216A3 (en) | 1995-07-05 |
| AU669603B2 (en) | 1996-06-13 |
| JPH075178A (en) | 1995-01-10 |
| AU5776294A (en) | 1994-09-22 |
| EP0616216A2 (en) | 1994-09-21 |
| US5372948A (en) | 1994-12-13 |
| JP3447360B2 (en) | 2003-09-16 |
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| EEER | Examination request | ||
| FZDE | Discontinued | ||
| FZDE | Discontinued |
Effective date: 20000308 |