Methods and test devices for determination of glycated haemoglobin

• US 6,399,293 B1
• Filed: 09/13/1999
• Issued: 06/04/2002
• Est. Priority Date: 09/21/1998
• Status: Expired due to Term

• Current Assignee: Roche Diagnostics GmbH (Roche Holding AG)
• Original Assignee: Roche Diagnostics GmbH (Roche Holding AG)
• Inventors: Horn, Carina, Pachl, Rudolf
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US09/395,228
• Time in Patent Office: 995 Days
• Field of Search: 435/4, 435/7.92, 435/7.95, 435/25, 435/970, 422/56-58, 422/60, 436/66, 436/67, 436/169, 436/177, 436/539, 436/828
• US Class Current: 435/4
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/54391 based on vertical flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/723 Glycosylated haemoglobin
• CPC Class Codes: G01N 33/726 Devices
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

This invention relates to a test element and a method for determining the ratio of glycated to non-glycated haemoglobin in a sample, for example, in a blood sample that is provided as a fresh sample, a dried sample or a haemolysed sample. The test element can include different zones for receiving the sample, serving as a depot for reagents such as non-immobilized signal-generating molecules or haemolysis reagents, separating reagents from the sample, and detecting heamoglobin. The zones are provided on a porous matrix and in fluid contact with each other. The method uses the test element and determines the ratio of glycated to non-glycated haemoglobin in a sample, by contacting the sample with an excess of signal generating molecules to bind to the glycated haemoglobin. The non-bound signal generating molecules are separated from the sample, and the amount or concentration of haemoglobin and/or glycated haemoglobin bound signal generating molecules are determined. The signal generating molecules can be selected to be a direct label, an enzyme label or a radiolabel, which can be attached to the glycated haemoglobin via a boronic acid group or another specific binding partner or antibody.

Claims

1. An analytical test element for determining the ratio of glycated to non-glycated haemoglobin in a sample, the test element comprising:

a sample application zone;

a reagent zone in which non-immobilized signal-generating molecules are present in excess relative to an expected amount of glycated haemoglobin in the sample or which contains one or more haemolysis reagents to release cell-bound haemoglobin or both a fraction of the non-immobilized signal-generating molecules dissolve into the sample and bind to the glycated haemoglobin in the sample;

a separation zone for separating signal-generating molecules that are dissolved in the sample but not bound to glycated haemoglobin from the sample which contains signal-generating molecules bound to glycated haemoglobin and non-glycated haemoglobin; and

a detection zone for detecting the amount of non-glycated haemoglobin and for detecting the amount of signal-generating molecules bound to glycated haemoglobin, wherein one or more of the zones are located on or in at least one porous matrix material and are in contact with one another enabling liquid transfer or can be brought into such contact enabling liquid transfer from one another.

2. The analytical test element of claim 1 wherein the separation zone is a porous matrix material comprising positively charged groups.

3. The analytical test element of claim 1 wherein a retardation zone is located between the reagent zone and the separation zone.

4. The analytical test element of claim 1 wherein the at least one porous matrix material is mounted on an inert support.

5. The analytical test element of claim 1 wherein the sample comprises non-immoblized signal generating molecules prior to introduction in the sample application zone.

6. An analytical test element comprising:

a sample application zone for receiving a sample comprising haemoglobin and glycated haemoglobin;

a reagent zone for contacting the sample with a reagent comprising non-immobilized signal-generating molecules or haemolysis reagents or both, said non-immobilized signal generating molecules provided in an excess amount sufficient to convert substantially all of the glycated haemoglobin in the sample to a detectable complex comprising signal generating molecules bound to glycated haemoglobin;

a separation zone comprising a porous matrix material including positively charged groups provided to retain any non-immoblized signal generating molecules in the sample; and

a detection zone for detecting the amount of detectable complex or a total amount of haemoglobin or both in the sample, wherein one or more of the zones are located on or in at least one porous matrix material and are in contact with one another or can be brought into contact with one another to allow liquid transfer through the zones.

7. The analytical test element of claim 6 wherein the detection zone is adapted to detect a total amount of glycated haemoglobin, non-glycated haemoglobin and detectable complex in the sample.

8. The analytical test element of claim 6 wherein the sample comprises non-immoblized signal generating molecules prior to introduction in the sample application zone.

9. The analytical test element of claim 6 wherein the porous matrix material comprises a nylon membrane.

10. The analytical test element of claim 9 wherein the nylon membrane contains quarternary ammonium groups.

11. The analytical test element of claim 6 wherein a retardation zone is located between the reagent zone and the separation zone.

12. The analytical test element of claim 6 comprising at least one porous matrix material mounted on an inert support.

13. The analytical test element of claim 6 wherein the reagent zone comprises at least one buffer substance.

14. The analytical test element of claim 6 wherein the signal generating molecules comprise a conjugate of one or several dihydroxyboryl residues or salts thereof, which are bound to a signal-generating label.

15. The analytical test element of claim 6 wherein the signal generating molecules comprise boronic acid conjugate compounds with an absorption maximum of not less than 600 nm or salt thereof wherein the compounds have the general Formula I