Reaction cassette for preforming sequential analytical assays by noncentrifugal and noncapillary manipulations

Claim 3

The method of claim 1 wherein said flow disrupting means is situated along said reaction channel a sufficient distance from said reagent zone that said liquid mixture can be in contact with said reagent zone without simultaneously being in contact with said flow disrupting means.

Claim 4

The method of claim 1 wherein said flow disrupting means and said reagent zone are situated sufficiently proximate to each other that said liquid mixture can be in contact with said flow disrupting means and said reagent zone simultaneously.

Claim 5

The method of claim 1 wherein said flow disrupting means comprises an obstruction or a point of convergence in said reaction channel configured such that the gravity flow of a liquid is redirected upon contact of said liquid therewith.

Claim 6

The method of claim 1 wherein said flow disrupting means is a corner in an outer perimeter wall portion of said reaction channel which forms an angle of between about 75 degrees and about 105 degrees.

Claim 7

The method of claim 5 wherein said corner in said outer perimeter wall portion of said reaction channel forms and angle of about 90 degrees.

Claim 8

The method of claim 1 wherein the speed of rotation of said reaction cassette about said horizontal axis used to transport said liquid mixture along said reaction channel and into contact with said reagent and flow disrupting means is sufficiently slow to be essentially noncentrifugal and due substantially only to gravitational force.

Claim 9

The method of claim 7 wherein said speed of rotation is between about 1 r.p.m. and about 60 r.p.m.

Claim 10

The method of claim 1 wherein the maximum speed of rotation of said reaction cassette during its oscillation to agitate the liquid mixture in contact with said flow disrupting means is between about 15 r.p.m. and about 40 r.p.m.

Claim 11

The method of claim 1 wherein said reagent zone is incorporated with a dried form of said analytical reagent.

Claim 12

The method of claim 1 wherein said reaction cassette also comprises means for introducing a liquid analytical reagent into said reaction channel.

Claim 13

The method of claim 1 wherein said reaction cassette also comprises means for introducing a liquid test sample into said reaction channel through said inlet means.

Claim 14

The method of claim 1 wherein said reaction channel further comprises a viewing zone in open liquid flow communication with said reaction zone.

Claim 15

The method of claim 13 comprising the step of rotating said reaction cassette about said horizontal axis whereby said liquid mixture is transported by gravity along said reaction channel into said reaction mixture viewing zone.

Claim 16

The method of claim 13 wherein a second outer perimeter wall portion of said reaction channel is configured as a corner forming an angle of between about 75 degrees and about 105 degrees, and wherein said corner serves as said reaction mixture viewing zone.

Claim 17

The method of claim 1 wherein said reaction channel comprises a plurality of reagent zones incorporated with a plurality of analytical reagents, and wherein rotation of the reaction cassette about its horizontal axis is used to bring the liquid mixture in the reaction channel into contact with each such reagent zones by gravity flow.

Claim 18

The method of claim 16 wherein after being brought into contact with a reagent zone, the liquid mixture is agitated in contact with flow disrupting means by oscillation of the reaction cassette about its horizontal axis.

Claim 19

An immunoturbidometric method of determining the relative amount of glycated hemoglobin in a whole blood test sample, said method comprising the steps of: (a) providing a capillary holder that comprises a capillary sampling tube, (b) drawing said whole blood test sample into the capillary sampling tube in said capillary holder, (c) providing a reaction cassette having a substantially horizontal axis of rotation, said reaction cassette comprising (1) a reaction channel situated in the plane of rotation about said horizontal axis and comprising (i) an outer perimeter wall having first and second corners which form angles in said perimeter wall of between about 75 degrees and about 105 degrees, (ii) a first reagent zone proximal to said first corner and incorporated with an oxidant capable of converting hemoglobin into its met-hemoglobin form, (iii) a second reagent zone incorporated with an antibody-particle reagent, and (iv) a third reagent zone incorporated with an agglutinator reagent; (2) liquid delivery means for introducing an aqueous solution of hemoglobin denaturant into the reaction channel, and (3) an inlet port engageable with the capillary holder to position the capillary sampling tube proximate to said first corner of the reaction channel, (d) engaging said capillary holder in said inlet port of the reaction vessel, (e) activating said liquid delivery means to release the denaturant solution into the reaction channel of the reaction vessel, (f) rotating the reaction cassette about its horizontal axis to transport by gravity the denaturant solution along the reaction channel to said first corner of the reaction channel and into contact with the capillary sampling tube and said oxidant in said first reagent zone, (g) oxcillating the reaction cassette about its horizontal axis to agitate the denaturant solution in said first corner in contact with said capillary sampling tube and the oxidant-incorporated first reagent zone sufficiently to extract the blood sample from the capillary tube and to form a reaction mixture with the denaturant and oxidant, (h) measuring the absorbance of met-hemoglobin formed in the resulting liquid mixture, (i) rotating the reaction cassette about its horizontal axis to transport by gravity the liquid reaction mixture along the reaction channel into contact with said second and third reagent zones and said second corner, (j) oscillating the reaction cassette about its horizontal axis to agitate the liquid reaction mixture in said second corner in contact with the second and third reagent zones sufficiently to thoroughly mix said liquid mixture and the antibody-particle and agglutinator reagents, (k) measuring the absorbance of the resulting liquid mixture and relating the ratio of the absorbance measured in step (h) to such final absorbance measurement as a function of the relative amount of glycatedhemoglobin in the blood sample tested.

Claim 20

An immunoturbidometric method of determining the relative amount of glycated hemoglobin in a whole blood test sample, said method comprising the steps of: (a) providing a capillary holder that comprises a capillary sampling tube, (b) drawing said whole blood test sample into the capillary sampling tube in said capillary holder, (c) providing a reaction cassette having a substantially horizontal axis of rotation, said reaction cassette comprising (1) a reaction channel situated in the plane of rotation about said horizontal axis and comprising (i) an outer perimeter wall having first and second corners which form angles in said perimeter wall of between about 75 degrees and about 105 degrees, (ii) a first reagent zone proximal to said first corner and incorporated with an oxidant capable of converting hemoglobin into its met-hemoglobin form, (iii) a second reagent zone incorporated with an antibody-particle reagent, and (iv) a third reagent zone incorporated with an agglutinator reagent; (2) liquid delivery means for introducing an aqueous solution of hemoglobin denaturant into the reaction channel, and (3) an inlet port engageable with the capillary holder to position the capillary sampling tube proximate to said first corner of the reaction channel, (d) engaging said capillary holder in said inlet port of the reaction vessel, (e) activating said liquid delivery means to release the denaturant solution into the reaction channel of the reaction vessel, (f) rotating the reaction cassette about its horizontal axis to transport by gravity the denaturant solution along the reaction channel to said first corner of the reaction channel and into contact with the capillary sampling tube and said oxidant in said first reagent zone, (g) oxcillating the reaction cassette about its horizontal axis to agitate the denaturant solution in said first corner in contact with said capillary sampling tube and the oxidant-incorporated first reagent zone sufficiently to extract the blood sample from the capillary tube and to form a reaction mixture with the denaturant and oxidant, (h) measuring the absorbance of met-hemoglobin formed in the resulting liquid mixture, (i) rotating the reaction cassette about its horizontal axis to transport by gravity the liquid reaction mixture along the reaction channel into contact with said second and third reagent zones and said second corner, (j) oscillating the reaction cassette about its horizontal axis to agitate the liquid reaction mixture in said second corner in contact with the second and third reagent zones sufficiently to thoroughly mix said liquid mixture and the antibody-particle and agglutinator reagents, (k) measuring the absorbance of the resulting liquid mixture and relating the ratio of the absorbance measured in step (h) to such final absorbance measurement as a function of the relative amount of glycatedhemoglobin in the blood sample tested.

Claim 21

The method of claim 18 wherein said first or second corner in the reaction channel provides a viewing zone for measuring the absorbance of liquid mixtures in the reaction channel, and wherein the liquid mixtures are transported by gravity to such corner by rotation of the reaction vessel about its horizontal axis.

Claim 22

The method of claim 18 wherein said oxidant comprises a ferricyanide salt.

Claim 23

The method of claim 18 wherein said liquid denaturant comprises a thiocyanate salt.

Claim 24

The method of claim 18 wherein said antibody particle reagent comprises an antibody, or fragment thereof, to the glycated N-terminal peptide sequence in the beta-subunit of hemoglobin, which antibody or fragment is immobilized to a water suspendable particle.

Claim 25

The method of claim 22 wherein said water suspendable particle is a latex particle.

Claim 26

The method of claim 18 wherein said agglutinator reagent comprises a plurality of epitopic binding sites for said antibody reagent.

Claim 27

The method of claim 24 wherein said epitopic sites are glycated peptide residues corresponding to the glycated N-terminal peptide sequence of hemoglobin.

Claim 28

The method of claim 18 wherein said liquid delivery means is centrally located in said reaction cassette.

Claim 29

The method of claim 18 wherein said liquid delivery means is in the form of a syringe.

Claim 30

The method of claim 18 wherein said liquid delivery means is in the form of a reservoir body closed by a removable, fluid tight seal.

Claim 31

The method of claim 18 wherein said corners each form an angle of about 90 degrees.

Claim 32

The method of claim 18 wherein the speed of rotation of said reaction cassette about said horizontal axis used in steps (f) and (i) to transport liquid along the reaction channel is sufficiently slow to be essentially noncentrifugal and due substantially only to gravitational force.

Claim 33

The method of claim 30 wherein said speed of rotation is between about 1 r.p.m. and about 60 r.p.m.

Claim 34

The method of claim 18 wherein the maximum speed of rotation of said reaction vessel during its oscillation to agitate the reaction liquid in performing steps (g) and (j) is between about 15 r.p.m. and about 40 r.p.m.

Claim 35

An analytical reaction cassette for performing analytical reactions to determine an analyte in a liquid test sample, said reaction cassette having a substantially horizontal axis of rotation and comprising: (1) inlet means for introducing a liquid test sample into said reaction cassette, and (2) a reaction channel in open liquid communication with said inlet means and comprising (i) a reagent zone incorporated with an analytical reagent which interacts with said analyte to produce a detectable response as a function of the analyte, and (ii) means for disrupting the flow of a liquid by gravity along said reaction channel sufficient to agitate said liquid upon contact therewith, whereby a liquid disposed in said reaction channel is transported by gravity along said reaction channel by rotating said reaction vessel about said horizontal axis and thereby brought into contact with said reagent zone and said flow disrupting means.

Claim 36

An analytical reaction cassette for performing analytical reactions to determine an analyte in a liquid test sample, said reaction cassette having a substantially horizontal axis of rotation and comprising: (1) inlet means for introducing a liquid test sample into said reaction cassette, and (2) a reaction channel in open liquid communication with said inlet means and comprising (i) a reagent zone incorporated with an analytical reagent which interacts with said analyte to produce a detectable response as a function of the analyte, and (ii) means for disrupting the flow of a liquid by gravity along said reaction channel sufficient to agitate said liquid upon contact therewith, whereby a liquid disposed in said reaction channel is transported by gravity along said reaction channel by rotating said reaction vessel about said horizontal axis and thereby brought into contact with said reagent zone and said flow disrupting means.

Claim 37

The reaction cassette of claim 33 wherein said flow disrupting means is situated along said reaction channel a sufficient distance from said reagent zone that a liquid can be in contact with said reaction zone without simultaneously being in contact with said flow disrupting means.

Claim 38

The reaction cassette of claim 33 wherein said flow disrupting means and said reagent zone are situated sufficiently proximate to each other that a liquid can be in contact with said flow disrupting means and said reagent zone simultaneously.

Claim 39

The reaction cassette of claim 33 wherein said flow disrupting means comprises an obstruction or a point of convergence in said reaction channel configured such that the gravity flow of a liquid is redirected upon contact of said liquid therewith.

Claim 40

The reaction cassette of claim 33 wherein said flow disrupting means is a corner in an outer perimeter wall portion of said reaction channel which forms an angle of between about 75 degrees and about 105 degrees.

Claim 41

The reaction cassette claim 37 wherein said corner in said outer perimeter wall portion of said reaction channel forms an angle of about 90 degrees.

Claim 42

The reaction cassette of claim 33 wherein said reagent zone is incorporated with a dried form of said analytical reagent.

Claim 43

The reaction cassette of claim 33 also comprising means for introducing a liquid analytical reagent into said reaction channel.

Claim 44

The reaction cassette of claim 33 also comprising means for introducing a liquid test sample into said reaction channel through said inlet means.

Claim 45

The reaction cassette of claim 33 wherein said reaction channel further comprises a reaction mixture viewing zone in open liquid flow communication with said reaction zone.

Claim 46

The reaction cassette of claim 42 wherein a second outer perimeter wall portion of said reaction channel is configured as a corner forming an angle of between about 75 degrees and about 105 degrees, and wherein said corner serves as said reaction mixture viewing zone.

Claim 47

The reaction cassette of claim 33 wherein said reaction channel comprises a plurality of reagent zones incorporated with a plurality of analytical reagents.

Claim 48

An immunoturbidimetric reaction cassette kit for determining the relative amount of glycated hemoglobin in a whole blood test sample, said kit comprising: (A) a capillary holder that comprises a capillary sampling tube, and (B) a reaction cassette body having a substantially horizontal axis of rotation and comprising (1) a reaction channel situated in the plane of rotation about said horizontal axis and comprising (i) an outer perimeter wall having first and second corners which form angles in said perimeter wall of between about 75 degrees and about 105 degrees, (ii) a first reagent zone proximal to said first corner and incorporated with an oxidant capable of converting hemoglobin into its met-hemoglobin form, (iii) a second reagent zone incorporated with an antibody-particle reagent, and (iv) a third reagent zone incorporated with an agglutinator reagent, whereby a liquid disposed in the reaction channel is transported by gravity along said reaction channel and into contact with said corners and said reagent zones by rotation of the reaction cassette about its horizontal axis, (2) liquid delivery means for introducing an aqueous solution of hemoglobin denaturant into the reaction channel, and (3) an inlet port engageable with the capillary holder to position the capillary sampling tube proximate to said first corner of the reaction channel.

Claim 49

An immunoturbidimetric reaction cassette kit for determining the relative amount of glycated hemoglobin in a whole blood test sample, said kit comprising: (A) a capillary holder that comprises a capillary sampling tube, and (B) a reaction cassette body having a substantially horizontal axis of rotation and comprising (1) a reaction channel situated in the plane of rotation about said horizontal axis and comprising (i) an outer perimeter wall having first and second corners which form angles in said perimeter wall of between about 75 degrees and about 105 degrees, (ii) a first reagent zone proximal to said first corner and incorporated with an oxidant capable of converting hemoglobin into its met-hemoglobin form, (iii) a second reagent zone incorporated with an antibody-particle reagent, and (iv) a third reagent zone incorporated with an agglutinator reagent, whereby a liquid disposed in the reaction channel is transported by gravity along said reaction channel and into contact with said corners and said reagent zones by rotation of the reaction cassette about its horizontal axis, (2) liquid delivery means for introducing an aqueous solution of hemoglobin denaturant into the reaction channel, and (3) an inlet port engageable with the capillary holder to position the capillary sampling tube proximate to said first corner of the reaction channel.

Claim 50

The reaction kit of claim 45 wherein said liquid delivery means is centrally located in said reaction cassette.

Claim 51

The reaction kit of claim 45 wherein said liquid delivery means is in the form of a syringe.

Claim 52

The reaction kit of claim 45 wherein said liquid delivery means is in the form of a reservoir body closed by a removable, fluid tight seal.

Claim 53

The reaction kit of claim 45 wherein one of said corners serves as a reaction viewing zone from which a detectable response can be detected and measured.

Claim 54

The reaction kit of claim 45 wherein said oxidant, said antibody particle reagent, and said agglutinator reagent are incorporated as substantially dry, dissolvable forms thereof.

Claim 55

The reaction kit of claim 45 wherein said oxidant comprises a ferricyanide salt.

Claim 56

The reaction kit of claim 45 wherein said liquid denaturant comprises a thiocyanate salt.

Claim 57

The reaction kit of claim 45 wherein said antibody particle reagent comprises an antibody, or fragment thereof, to the glycated N-terminal peptide sequence in the beta-subunit of denatured hemoglobin, which antibody or fragment is immobilized to a suspendable particle.

Claim 58

The reaction kit of claim 53 wherein said suspendable particle is a water suspendable latex particle.

Claim 59

The reaction kit of claim 45 wherein said agglutinator reagent comprises a plurality of epitopic binding sites for said antibody reagent.

Claim 60

The reaction kit of claim 55 wherein said agglutinator reagent comprises glycated peptide residues of glycated hemoglobin.

Claim 61

The reaction kit of claim 45 wherein said corners each form an angle of 90 degrees.