A method for detecting abnormal results in immunoassays caused by insufficient delivery of polyhapten reagents.

Abstract

To disclose a method of detecting aberrant results caused by delivery issues for a polyhapten reagent in the context of immunoassays.SOLUTION: Operation is added to an assay parameter for estimating an equipment signal when a polyhapten reagent is delivered. This method predicts a signal at 0 hour by using regression of two antecedent readings of the time, subtracts a signal contribution from a sample and an antibody reagent in a reaction mixture, and provides accurate monitoring of the signal by adding the polyhapten reagent to reaction. By removing the signal contribution from the sample and the antibody reagent, equipment software can compare signals from the delivery of each test polyhapten reagent and can stand a flag to the test affected by a problem of reagent delivery. This method is effective because a change of the signal relative to the time is linear at the point of time when it is used.SELECTED DRAWING: Figure 1

Claims

1. A reagent kit for use in a method for detecting abnormal results caused by insufficient delivery of particle agglutination assay reagents used in immunoassays, wherein the method is (A) A step of reacting a biological sample suspected to contain the target analyte with a target analyte-specific binding partner in a reaction cuvette, thereby forming a soluble analyte / specific binding partner complex, (B) A step of adding a particle agglutination assay reagent to the reaction cuvette, wherein the particle agglutination assay reagent reacts with an excess of the target analyte-specific binding partner to form an insoluble particle agglutination assay reagent / target analyte-specific binding partner complex, (C) A step of irradiating the reaction cuvette with light, (D) A step of measuring absorbance values ​​at at least three wavelengths at multiple time points after the addition of the particle agglutination assay reagent, wherein the first wavelength detects the insoluble particle agglutination assay reagent / target analyte-specific binding partner complex by turbidimetry, the second wavelength detects proteins, and the third wavelength functions as a blank, (E) A step of estimating the absorbance value at the time of delivery of the particle agglutination assay reagent by using the regression of absorbance values ​​measured at the second and third wavelengths at two time points after the addition of the particle agglutination assay reagent, (F) If the estimated absorbance value of the particle agglutination assay reagent at the time of delivery differs from the predicted value by an established flag constant, the concentration value of the target analyte obtained by the immunoassay is flagged as unacceptable. The reagent kit comprises a particle agglutination assay reagent that can specifically bind to a target analyte-specific binding partner, and the reagent kit comprises at least one target analyte-specific binding partner and at least one particle agglutination assay reagent.

2. The target analytes are glycated hemoglobin (HbA1c), albumin, human chorionic gonadotropin (hCG), ferritin, growth hormone, prolactin, thyroglobulin (Tg), C-reactive protein (CRP), rheumatoid factor (RF), gentamicin, tobramycin, CRP, digoxin, amikacin, caffeine, carbamazepine, digitoxin, The reagent kit according to claim 1, selected from the group consisting of disopyramide, ethosuxamide, lidocaine, lithium methotrexate, NAPA, phenobarbital, phenytoin, primidone, procainamide, quinidine, theophylline, tobramycin, valproic acid, and vancomycin.

3. The reagent kit according to claim 1, wherein the target analyte-specific binding partner is an antibody against the target analyte.

4. The reagent kit according to claim 3, wherein the target analyte is glycated hemoglobin (HbA1c), the target analyte-specific binding partner is an anti-HbA1c antibody, and the particle agglutination assay reagent comprises a plurality of HbA1c epitopes.

5. The reagent kit according to claim 1, wherein the first wavelength is in the range of 300 nm to 650 nm, the second wavelength is in the range of 190 nm to 300 nm, and the third wavelength is in the range of 650 nm to 850 nm.

6. The reagent kit according to claim 5, wherein the first wavelength is 340 nm, the second wavelength is 293 nm, and the third wavelength is 700 nm.

7. The reagent kit according to claim 1, wherein in step (E), the first of two time points after the addition of the particle agglutination assay reagent is 7.2 seconds after the addition of the particle agglutination assay reagent, and the second of the two time points is 7.2 seconds after the first time point.

8. The reagent kit according to claim 1, wherein the biological sample is selected from the group consisting of urine, whole blood or any part thereof, whole blood cells or lysed blood cells, saliva, sputum, cerebrospinal fluid, intestinal fluid, intraperitoneal fluid, cystic fluid, sweat, interstitial fluid, tears, mucus, bladder lavage fluid, semen, and combinations thereof.

9. A reagent kit for use in a method for detecting abnormal results caused by insufficient delivery of particle agglutination assay reagents used in glycated hemoglobin (HbA1c) immunoassays, wherein the method is: (A) A step of reacting a biological sample suspected to contain a target analyte including HbA1c with an anti-HbA1c antibody in a reaction cuvette to form a soluble HbA1c-antibody complex, (B) A step of adding a particle agglutination assay reagent to the reaction cuvette, wherein the particle agglutination assay reagent reacts with an excess of anti-HbA1c antibody to form an insoluble particle agglutination assay reagent / anti-HbA1c antibody complex, (C) A step of irradiating the reaction cuvette with light, (D) A step of measuring absorbance values ​​at at least three wavelengths at multiple time points after the addition of the particle agglutination assay reagent, wherein the first wavelength is used to detect the insoluble particle agglutination assay reagent / anti-HbA1c antibody complex by turbidimetry, the second wavelength is used to detect proteins, and the third wavelength functions as a blank, (E) A step of estimating the absorbance value at the time of delivery of the particle agglutination assay reagent by using the regression of absorbance values ​​measured at the second and third wavelengths at two time points after the addition of the particle agglutination assay reagent, (F) If the estimated absorbance value of the particle agglutination assay reagent at the time of delivery differs from the predicted value by an established flag constant, the concentration value of the target analyte obtained by the immunoassay is flagged as unacceptable. The reagent kit comprises at least one anti-HbA1c antibody and at least one particle agglutination assay reagent.

10. The reagent kit according to claim 9, wherein the first wavelength is in the range of 300 nm to 650 nm, the second wavelength is in the range of 190 nm to 300 nm, and the third wavelength is in the range of 650 nm to 850 nm.

11. The reagent kit according to claim 10, wherein the first wavelength is 340 nm, the second wavelength is 293 nm, and the third wavelength is 700 nm.

12. The reagent kit according to claim 9, wherein in step (E), the first of two time points after the addition of the particle agglutination assay reagent is 7.2 seconds after the addition of the particle agglutination assay reagent, and the second of the two time points is 7.2 seconds after the first time point.

13. The reagent kit according to claim 9, wherein the biological sample is selected from the group consisting of urine, whole blood or any part thereof, whole blood cells or lysed blood cells, saliva, sputum, cerebrospinal fluid, intestinal fluid, intraperitoneal fluid, cystic fluid, sweat, interstitial fluid, tears, mucus, bladder lavage fluid, semen, and combinations thereof.

14. A reagent kit for use in a method for detecting abnormal results caused by insufficient delivery of particle agglutination assay reagents used in glycated hemoglobin (HbA1c) immunoassays, wherein the method is: (A) A step of reacting a biological sample suspected to contain a target analyte including HbA1c with an anti-HbA1c antibody in a reaction cuvette to form a soluble HbA1c-antibody complex, (B) A step of adding a particle agglutination assay reagent to the reaction cuvette, wherein the particle agglutination assay reagent reacts with an excess of anti-HbA1c antibody to form an insoluble particle agglutination assay reagent / anti-HbA1c antibody complex, (C) A step of irradiating the reaction cuvette with light, (D) A step of measuring absorbance values ​​at at least three wavelengths at multiple time points after the addition of the particle aggregation assay reagent, (i) The first wavelength is in the range of 300 nm to 650 nm when the insoluble particle agglutination assay reagent / anti-HbA1c antibody complex is detected by turbidimetry, (ii) The second wavelength detects proteins and is in the range of 190 nm to 300 nm. (iii) The process described above, wherein a third wavelength functions as a blank and is in the range of 650 nm to 850 nm, (E) A step of estimating the absorbance value at the time of delivery of the particle agglutination assay reagent by using the regression of absorbance values ​​measured at the second and third wavelengths at two time points after the addition of the particle agglutination assay reagent, (F) If the estimated absorbance value of the particle agglutination assay reagent at the time of delivery differs from the predicted value by an established flag constant, the concentration value of the target analyte obtained by the immunoassay is flagged as unacceptable. The reagent kit comprises at least one anti-HbA1c antibody and at least one particle agglutination assay reagent.

15. The reagent kit according to claim 14, wherein the first wavelength is 340 nm, the second wavelength is 293 nm, and the third wavelength is 700 nm.

16. The reagent kit according to claim 14, wherein in step (E), the first of two time points after the addition of the particle agglutination assay reagent is 7.2 seconds after the addition of the particle agglutination assay reagent, and the second of the two time points is 7.2 seconds after the first time point.

17. The reagent kit according to claim 14, wherein the biological sample is selected from the group consisting of urine, whole blood or any part thereof, whole blood cells or lysed blood cells, saliva, sputum, cerebrospinal fluid, intestinal fluid, intraperitoneal fluid, cystic fluid, sweat, interstitial fluid, tears, mucus, bladder lavage fluid, semen, and combinations thereof.

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