US 12,247,987 B2
Methods for detection of aberrant results caused by incomplete dispersion of immunoassay reagent
Tie Wei, Wilmington, DE (US)
Assigned to Siemens Healthcare Diagnostics Inc., Tarrytown, NY (US)
Appl. No. 17/250,347
Filed by Siemens Healthcare Diagnostics Inc., Tarrytown, NY (US)
PCT Filed Jun. 28, 2019, PCT No. PCT/US2019/039790
§ 371(c)(1), (2) Date Jan. 8, 2021,
PCT Pub. No. WO2020/014012, PCT Pub. Date Jan. 16, 2020.
Claims priority of provisional application 62/697,651, filed on Jul. 13, 2018.
Prior Publication US 2021/0270849 A1, Sep. 2, 2021
Int. Cl. G01N 33/72 (2006.01)
CPC G01N 33/723 (2013.01) 23 Claims
 
1. A method of detecting aberrant results caused by incomplete dispersion of a polyhapten reagent used in a turbidimetric HbA1c immunoassay, the method comprising the steps of:
(A) reacting, within a reaction cuvette, a biological sample suspected of containing a target analyte comprising glycated hemoglobin (HbA1c) with an anti-HbA1c antibody to the target analyte, thereby forming a soluble HbA1c-antibody complex;
(B) adding a polyhapten reagent to the reaction cuvette, wherein the polyhapten reagent reacts with excess anti-HbA1c antibody to form an insoluble polyhapten/target analyte-specific binding partner polyhapten/anti-HbA1c antibody complex;
(C) irradiating the reaction cuvette with light at a first wavelength and a second wavelength;
(D) turbidimetrically detecting the insoluble polyhapten/target analyte-specific binding partner polyhapten/anti-HbA1c antibody complex and undispersed polyhapten reagent in the reaction mixture by measuring an absorbance value at the first wavelength and an absorbance value at the second wavelength;
(E) comparing the measured first and second wavelength absorbance values to predicted values of an established regression obtained from first and second wavelength absorbances at known HbA1c control values for the target analyte having different concentrations during assay calibration; and
(F) flagging, as unacceptable, a concentration value for the target analyte HbA1c obtained by a separate algorithm if the measured second wavelength absorbance value exceeds the predicted value by an established flag constant.
 
12. A method of detecting aberrant results caused by incomplete dispersion of a polyhapten reagent used in a turbidimetric HbA1c immunoassay, the method comprising the steps of:
(A) reacting, within a reaction cuvette, a biological sample suspected of containing a target analyte comprising glycated hemoglobin (HbA1c) with an anti-HbA1c antibody to the target analyte, thereby forming a soluble HbA1c antibody HbA1c/anti-HbA1c antibody complex;
(B) adding a polyhapten reagent to the reaction cuvette, wherein the polyhapten reagent reacts with excess anti-HbA1c antibody to form an insoluble polyhapten/target analyte-specific binding partner polyhapten/anti-HbA1c antibody complex;
(C) irradiating the reaction cuvette with light at a first wavelength and a second wavelength;
(D) turbidimetrically detecting the insoluble polyhapten/target analyte-specific binding partner polyhapten/anti-HbA1c antibody complex and undispersed polyhapten reagent in the reaction mixture by measuring an absorbance value at the first wavelength and an absorbance value at the second wavelength;
(E) comparing the measured first and second wavelength absorbance values to predicted values of an established regression obtained from first and second wavelength absorbances at known HbA1c control values for the target analyte having different concentrations during assay calibration;
(F) flagging and suppressing a concentration value for the target analyte HbA1c obtained by a separate algorithm if the measured second wavelength absorbance value exceeds the predicted value by an established flag constant; and
(G) reporting a target analyte an HbA1c concentration if the measured second wavelength absorbance value does not exceed the predicted value by the established flag constant.