Procedure for the determination of the reaction lag phase in an analyte-dependent reaction

• US 7,981,622 B2
• Filed: 04/16/2008
• Issued: 07/19/2011
• Est. Priority Date: 04/17/2007
• Status: Expired due to Fees

• Current Assignee: Siemens Healthcare Diagnostics Products GmbH (Siemens AG)
• Original Assignee: Siemens Healthcare Diagnostics Products GmbH (Siemens AG)
• Inventors: Henckel, Thilo, Weyl, Andreas
• Primary Examiner(s): Prouty; Rebecca
• Assistant Examiner(s): MARTIN, PAUL C
• Application Number:
  US12/081,470
• Publication Number:
  US 20080261254A1
• Time in Patent Office: 1,189 Days
• Field of Search: None
• US Class Current: 435/7.1
• CPC Class Codes: C12Q 1/56 involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
• CPC Class Codes: G01N 33/557 using kinetic measurement, i.e. time rate of progress of an antigen-antibody interaction
• CPC Class Codes: G01N 33/86 involving blood coagulating time or factors, or their receptors

Abstract

The invention is in the field of analytical technique and relates to an improved procedure for determining the concentration or activity of an analyte, an analyte-dependent detection reaction being set in motion and the reaction kinetics determined being evaluated. The procedure makes possible an individual determination of the lag phase of the reaction kinetics for each measurement.

Claims

1. A method for determining the concentration or activity of an analyte in a sample, the method comprising:

mixing a sample with at least one reagent, whereby an analyte-dependent reaction is set in motion;

measuring a signal (x) changing over time (t) as a result of the analyte-dependent reaction and storing a resultant signal-time curve;

determining a maximum increase in the signal-time curve;

determining a reaction lag phase of the signal-time curve; and

determining a concentration or activity of the analyte by means of at least one parameter of the signal-time curve, wherein the parameter occurs after the reaction lag phase in terms of time, and

wherein determining the reaction lag phase includes determining a period of time from a time of a first signal (t₀) up to a time t_lag,

by determining a time t_max of the maximum increase in the signal-time curve; and

subsequently determining from the time t_max, the time t_lag, where t_lag is smaller than t_max and where t_lag corresponds to the time at which the increase in the signal-time

curve for the first time reaches a threshold value and where the threshold value corresponds to a test-specific, predetermined fraction of the maximum increase in the signal-time curve.

2. The method of claim 1, wherein the threshold value corresponds to a fraction of 1 to 45% of the maximum increase in the signal-time curve.

3. The method of claim 1, wherein determining the concentration or activity of an analyte includes determining the concentration or activity of an analyte in a biological sample selected from the group consisting of whole blood, plasma, serum, urine, feces, saliva or cerebrospinal fluid.

4. The method of claim 1, wherein the analyte-dependent reaction is an antigen-antibody reaction or an enzyme-substrate reaction.

5. The method of claim 1, wherein measuring a signal (x) includes measuring a signal selected from the group consisting of scattered light, turbidity, fluorescence and chemiluminescence.

6. The method of claim 1, wherein the method for determining the concentration or activity of an analyte includes determining the concentration or activity of thrombin.

7. The method of claim 6, further comprising mixing the sample with a reagent for the activation of prothrombin to thrombin.

8. The method of claim 7, where the sample is mixed with a reagent comprising a thrombin substrate.

9. The method of claim 8, wherein measuring the signal (x) changing over time (t) includes measuring a signal (x) changing over time (t) as a result of the thrombin-dependent substrate cleavage and storing the resulting signal-time curve.

10. The method of claim 6, wherein the method for determining the concentration or activity of thrombin further comprises determining the formation and inhibition of endogenous thrombin.

11. The method of claim 1, further comprising determining a clotting time.

12. The method of claim 11, wherein determining the clotting time includes determining a clotting time selected from the group consisting of prothrombin time, activated partial thromboplastin time, thrombin time, batroxobin time and ecarin time.

13. The method of claim 1, wherein the at least one parameter is at least one of a maximum reaction rate, a time at which the maximum reaction rate occurs, an area under a first derivative of the signal-time curve, a time at which the signal (x) exceeds a predetermined signal, and a signal which is achieved at a predetermined time.

14. The method of claim 1, wherein the threshold value is determined by calculating a preliminary reaction lag phase and comparing the preliminary reaction lag phase to an expected lag phase over a number of tests.

15. The method of claim 1, wherein the maximum increase in the signal time curve is a slope of the signal time curve.