Methods of blood sample suspension

Patent metadata

Field	Value
Publication number	US-12474356-B2
Application number	US-202017310233-A
Country	US
Kind code	B2
Filing date	Feb 3, 2020
Priority date	Feb 6, 2019
Publication date	Nov 18, 2025
Grant date	Nov 18, 2025

Abstract

Disclosed herein are methods for rapid detection of suspension of a blood sample from a subject and methods for improving the processing time of a point of care blood analysis. Specifically, the disclosed methods provide a rapid detection of hemoglobin concentration, more specifically glycated hemoglobin A1c, and determination of sugar concentration in the blood sample from the subject. Further disclosed herein are methods for rapidly diagnosing diabetes in a subject.

First claim

The invention claimed is:

1 . An automated method for performing a blood sugar assay by a point of care immunoassay analyzer, the method comprising the following steps performed in the point of care immunoassay analyzer:
(a) generating a hemolysate mixture by mixing a blood sample from a subject with a buffer; (b) measuring an absorbance value of the buffer at a wavelength x (buffer x ) and an absorbance value of the buffer at a wavelength y (buffer y ); (c) measuring an absorbance value of the hemolysate mixture at a timepoint n at a wavelength x (hemo xn ) and at a wavelength y (hemo yn ); (d) determining a differential (dhemo xn ) between hemoxn and buffer x ; (e) determining a differential (dhemo yn ) between hemoyn, and buffer y ; (f) determining an absolute value (V n ) of the differential between dhemoxn and dhemo yn ; (g) repeating steps (c) to (e) for one additional consecutive timepoint n+1, wherein the timepoint n of step (c) becomes equal to n+1, to generate dhemo xn+1 and dhemo yn+1 ; (h) determining a differential between dhemoyn, and dhemo yn+1 , wherein yn and yn+1 relate to two consecutive measurements; (i) determining an absolute value (V N ) of the differential between dhemo yn , and dhemo yn+1 ; (j) determining an absolute value (V n+1 ) of the differential between dhemo xn+1 and dhemo yn+1 ; (k) assessing the absolute value V n , an absolute value (V n+1 )−(V n ), and the absolute value V N , wherein,
i. if step (k) is performed at less than a time t from a start of the mixing in step (a) and
a. the absolute value V n , is indicative of a glycated hemoglobin concentration of at least about 5 g/dL to about 21 g/dL, the absolute value (Vn+1)−(V n ) is about zero, and the absolute value V N is about zero, the blood sample is sufficiently mixed with the buffer for further analysis and the method proceeds to step (1); or
b. the absolute value of V n , is not indicative of a glycated hemoglobin concentration of at least about 5 g/dL to about 21g/dL, the absolute value of (Vn+1)−(V n ) is not about zero, or the absolute value of V N is not about zero, the hemolysate is mixed further and steps (g) to (j) are repeated;

ii. if step (k) is performed at time t from the start of the mixing in step (a) and
a. the absolute value V n , is indicative of a glycated hemoglobin concentration of at least about 5 g/dL to about 21 g/dL, the blood sample is sufficiently mixed with the buffer for further analysis and the method proceeds to step (1); or
b. the absolute value Vn is not indicative of a glycated hemoglobin concentration of at least about 5 g/dL to about 21 g/dL, the blood sample is not sufficiently mixed with the buffer for further analysis and steps (g) to (j) are repeated;

(l) analyzing the sufficiently mixed blood sample to detect a glycated hemoglobin concentration; (m) assessing a level of sugar in the blood sample based upon the glycated hemoglobin concentration so detected; and, (n) providing an output result for the blood sugar assay, the output result comprising the glycated hemoglobin concentration and the level of sugar in the blood sample; and wherein measuring the absorbance values in steps (b) and (c) and analyzing the sufficiently mixed blood sample to detect the glycated hemoglobin concentration of step (l) are accomplished with a spectrophotometer in the point of care immunoassay analyzer.

2 . The automated method of claim 1 , wherein the mixing of the blood sample comprises sonicating the blood sample.

3 . The automated method of claim 1 , wherein the buffer comprises lithium thiocyanate, digitonin, potassium ferricyanide and/or an anticoagulant selected from the group consisting of EDTA, heparin, citrate and fluoride/oxalate.

4 . The automated method of claim 1 , wherein the blood sample is in a vessel during the performance of at least one step, the vessel comprising a capillary tube, a capillary plate or a capillary membrane.

5 . The automated method of claim 1 , wherein the wavelength x is within the range of at least one set of wavelengths selected from the group consisting of:
a. 380 nm to 455 nm; b. 455 nm to 580 nm; and c. 580 nm to 680 nm; and, wherein the wavelength y is within a range of 700 nm to 2500 nm.

6 . The automated method of claim 1 , wherein the wavelengths x and y correspond respectively to 531 nm and 725 nm.

7 . The automated method of claim 1 , wherein step (c) is performed at least 2 seconds after step (d).

8 . The automated method of claim 1 , wherein, if the blood sample is indicated to be not sufficiently mixed in step (k)(ii)(b), the blood sample is replaced by another blood sample.

9 . The automated method of claim 1 , wherein the time t is about 60 seconds, about 70 seconds, about 80 seconds, about 90 seconds, about 100 seconds, about 110 seconds or about 120 seconds.

10 . The automated method of claim 1 , wherein the hemolysate is at least partially suspended.

11 . The automated method of claim 1 , wherein the blood sample is a human blood sample.

12 . The automated method of claim 1 , wherein the absolute value of (i) is about the absolute value of (0+/−0.002) or (−0.0002619+/−0.001349).

13 . The automated method of claim 1 , wherein the absolute value of (V n+1 )-(V n ) is about the absolute value of (0+/−0.002) or (−0.0006418+/−0.001758).

14 . The automated method of claim 1 , wherein the absoluate value V n assessed at step (k) is indicative of a glycated hemoglobin concentration of at least about one selected from the group consisting of: 5 g/dL and 7 g/dL.

15 . The automated method of claim 1 , wherein the glycated hemoglobin is hemoglobin A1c (HbA1c).

16 . An automated method for performing a blood sugar assay by a point of care immunoassay analyzer, the method comprising the following steps (a)-(f) performed in the point of care immunoassay analyzer:
(a) generating a hemolysate mixture by mixing a blood sample from a subject with a buffer; (b) taking multiple absorbance measurements of the mixed hemolysate mixture at instants of time with a spectrophotometer; (c) determining whether the hemolysate mixture is sufficiently mixed at different ones of the instants of times using the multiple absorbance measurements of the mixed hemolysate mixture, wherein the hemolysate mixture is determined to be sufficiently mixed when IΔ hemo -Δ bufter 1 is indicative of a hemoglobin concentration of at least about 5 g/dL+/−0.1 g/dL to about 21 g/dL, wherein Δ bufter is a differential of absorbance values of the buffer measured at a wavelength x and a wavelength y and Δ hemo is a differential of absorbance values of the hemolysate mixture measured at the wavelength x and the wavelength y; (d) responsive to the hemolysate mixture being insufficiently mixed, continue mixing the hemolysate mixture and repeat steps (b)-(c); and (e) responsive to the hemolysate mixture being sufficiently mixed, detecting a glycated hemoglobin concentration within the sufficiently mixed hemolysate mixture with the spectrophotometer; and (f) providing an output result comprising the glycated hemoglobin concentration.

17 . The method of claim 16 , wherein the output result is provided in less than 5 minutes thereby improving point of care blood analysis as compared to a standard point of care blood analysis.

Assignees

• Siemens Healthcare Diagnostics Inc

Inventors

• Kauffmann Aaron
• Zimmerle Chris

Classifications

• G01N21/31

  Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry {(G01N21/72 takes precedence)} · CPC title

• G01N1/38

  Diluting, dispersing or mixing samples · CPC title

• G01N21/314

  with comparison of measurements at specific and non-specific wavelengths (dual wavelength spectrometry G01J3/427) · CPC title

• A61B5/14546

  for measuring analytes not otherwise provided for, e.g. ions, cytochromes · CPC title

• G01N21/75

  Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated (systems in which material is burnt in a flame or plasma G01N21/72, G01N21/73) · CPC title