Detection of false results in assays

• US 8,084,223 B2
• Filed: 03/23/2009
• Issued: 12/27/2011
• Est. Priority Date: 03/23/2009
• Status: Active Grant

• Current Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Original Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Inventors: Wei, Tie Quan
• Primary Examiner(s): Gabel, Gail R
• Application Number:
  US12/409,302
• Publication Number:
  US 20100240073A1
• Time in Patent Office: 1,009 Days
• Field of Search: 435/7.1, 435/7.2, 435/7.25, 435/7.93, 436/520, 436/16, 436/17, 436/70, 436/175, 436/177, 436/522, 436/523, 436/526, 424/9.2
• US Class Current: 435/7.25
• CPC Class Codes: G01N 33/9493 Immunosupressants
• CPC Class Codes: Y10T 436/106664 Blood serum or blood plasma standard or control
• CPC Class Codes: Y10T 436/107497 Preparation composition [e.g., lysing or precipitation, etc.]
• CPC Class Codes: Y10T 436/25125 Digestion or removing interfering materials
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

Methods and reagents are disclosed for detecting a false result in an assay for determining a concentration of an analyte in a whole blood sample suspected of containing the analyte. The method comprises determining by means of the assay a concentration of the analyte utilizing a hemolyzed portion of the blood sample to obtain concentration value 1 and determining by means of the assay a concentration of the analyte utilizing a non-hemolyzed portion of the blood sample and multiplying the concentration times a hematocrit factor to obtain concentration value 2. A ratio of concentration value 1 divided by concentration value 2 is determined and is compared to a predetermined ratio of known reliability. If the ratio is less than the predetermined ratio, a false result is indicated.

Claims

1. A method for detecting a false result in an assay for determining a concentration of an erythrocytophilic analyte in a whole blood sample suspected of containing the erythrocytophilic analyte, the method comprising:

(a) determining by means of the assay a concentration of the erythrocytophilic analyte utilizing a hemolyzed portion of the blood sample to obtain concentration value 1,

(b) determining by means of the assay a concentration of the erythrocytophilic analyte utilizing a non-hemolyzed portion of the blood sample and multiplying the concentration times a hematocrit factor to obtain concentration value 2,

(c) determining a ratio of concentration value 1 divided by concentration value 2, and

(d) comparing the ratio from (c) to a predetermined ratio of known reliability wherein, if the ratio is less than the predetermined ratio, a false result is indicated.

2. The method of claim 1 wherein the predetermined ratio of known reliability is determined by a method that is independent of factors that produce a false result in the assay and wherein the predetermined ratio is a ratio of a concentration of the erythrocytophilic analyte in a hemolyzed portion of a different whole blood sample that contains the erythrocytophilic analyte divided by a concentration of erythrocytophilic analyte in a non-hemolyzed portion of the different whole blood sample.

3. The method according to claim 1 wherein the hematocrit factor is the following equation:

1/(1−

hematocrit % in decimal form).

4. The method according to claim 1 wherein the assay comprises:

(i) combining the blood sample of step (a) with reagents for determining the concentration of the erythrocytophilic analyte in the blood sample of step (a) wherein the reagents comprise at least one antibody for the erythrocytophilic analyte, and combining the blood sample of step (b) with reagents for determining the concentration of the erythrocytophilic analyte in the blood sample of step (b) wherein the reagents comprises at least one antibody for the erythrocytophilic analyte, and

(ii) measuring an amount of a complex comprising the erythrocytophilic analyte and the antibody for the erythrocytophilic analyte for the blood sample of step (a) and the blood sample of step (b) and relating the amount of the complex to the concentration of the erythrocytophilic analyte in the blood sample of step (a) and the blood sample of step (b).

5. The method according to claim 4 wherein the reagents in step (i) further comprise an analog of the erythrocytophilic analyte that comprises a label.

6. The method according to claim 4 wherein in step (i) a second antibody is added to the blood sample and reagents in step a) and step b) wherein the second antibody binds to the antibody for the erythrocytophilic analyte in the complex.

7. The method according to claim 6 wherein at least one of either the antibody for the erythrocytophilic analyte or the second antibody comprises a label.

8. The method according to claim 4 wherein one of the reagents comprises a label.

9. The method according to claim 4 wherein one of the reagents comprises a particle.

10. The method according to claim 4 wherein one of the reagents is an erythrocytophilic analyte analog linked to an enzyme label and one of the reagents is an antibody for the erythrocytophilic analyte attached to a magnetic particle.

11. A method for detecting a false result in an assay for determining a concentration of an erythrocytophilic analyte in a whole blood sample suspected of containing the erythrocytophilic analyte, the method comprising:

(a) determining by means of the assay a concentration of the erythrocytophilic analyte utilizing a hemolyzed portion of the blood sample to obtain concentration value 1,

(b) determining by means of the assay a concentration of the erythrocytophilic analyte utilizing a non-hemolyzed portion of the blood sample and multiplying the concentration times a hematocrit factor to obtain concentration value 2 wherein the hematocrit factor is the following equation;

1/(1−

hematocrit % in decimal form),

wherein the assay comprises;

(i) combining the blood sample of step (a) with reagents for determining the concentration of the erythrocytophilic analyte in the blood sample of step (a) wherein the reagents comprise at least one antibody for the erythrocytophilic analyte, and combining the blood sample of step (b) with reagents for determining the concentration of the erythrocytophilic analyte in the blood sample of step (b) wherein the reagents comprises at least one antibody for the erythrocytophilic analyte, and

(ii) measuring an amount of a complex comprising the erythrocytophilic analyte and the antibody for the erythrocytophilic analyte for the blood sample of step (a) and the blood sample of step (b) and relating the amount of the complex to the concentration of the erythrocytophilic analyte in the blood sample of step (a) and the blood sample of step (b),

(c) determining a ratio of concentration value 1 divided by concentration value 2, and

(d) comparing the ratio from (c) to a predetermined ratio of known reliability wherein, if the ratio is less than the predetermined ratio, a false result is indicated and wherein the predetermined ratio of known reliability is determined by a method that is independent of factors that produce a false result in the assay and wherein the predetermined ratio is a ratio of a concentration of the erythrocytophilic analyte in a hemolyzed portion of a different whole blood sample that contains the erythrocytophilic analyte divided by a concentration of erythrocytophilic analyte in a non-hemolyzed portion of the different whole blood sample.

12. The method according to claim 11 wherein the erythrocytophilic analyte is selected from the group consisting of tacrolimus, sirolimus and cyclosporin.

13. The method according to claim 11 wherein the erythrocytophilic analyte is an immunosuppressant drug.

14. The method according to claim 11 wherein the assay method is a heterogeneous assay method.

15. The method according to claim 11 wherein the reagents in step (i) further comprise an analog of the erythrocytophilic analyte that comprises a label.

16. The method according to claim 11 wherein in step (i) a second antibody is added to the blood sample and reagents in step a) and step b) wherein the second antibody binds to the antibody for the erythrocytophilic analyte in the complex.

17. The method according to claim 16 wherein at least one of either the antibody for the analyte or the second antibody comprises a label.

18. The method according to claim 11 wherein one of the reagents comprises a label.

19. The method according to claim 11 wherein one of the reagents comprises a particle.

20. The method according to claim 11 wherein one of the reagents is an erythrocytophilic analyte analog linked to an enzyme label and one of the reagents is an antibody for the erythrocytophilic analyte attached to a magnetic particle.