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Method for measuring glycosylated hemoglobin content

A glycosylated hemoglobin and percentage technology, which is applied in the field of medical immunity, can solve the problems of short stable period of antibody working solution, unfavorable daily use, and difficulty in obtaining low-temperature refrigerators, and achieves the effect of simplifying operation steps and prolonging effective use time.

Active Publication Date: 2010-12-15
SHENZHEN GOLDSITE DIAGNOSTICS
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AI Technical Summary

Problems solved by technology

There are two immunological methods commonly used clinically, one is the immunocompetitive inhibition method that needs to separately measure the concentration of total hemoglobin and glycosylated hemoglobin in blood samples, and then calculate the ratio of glycosylated hemoglobin to total hemoglobin, and the other It is a latex agglutination method that can directly measure the percentage of glycosylated hemoglobin. This method needs to prepare antibody working solution before use, and the prepared antibody working solution has a short stability period
[0004] The latex agglutination method that can directly measure the percentage of glycosylated hemoglobin, which is often used clinically, requires the preparation of antibody working solution before use, and the prepared antibody working solution has a short stability period, and generally fails in about 10-15 days. It can be stored for several months when frozen below -10°C, but it cannot be frozen and thawed repeatedly, which is not conducive to daily use, and low-temperature refrigerators are difficult to obtain

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  • Method for measuring glycosylated hemoglobin content
  • Method for measuring glycosylated hemoglobin content
  • Method for measuring glycosylated hemoglobin content

Examples

Experimental program
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Embodiment 1

[0024] The glycated hemoglobin detection reagent of this embodiment includes hemolysis, latex, antibody A reagent and antibody B reagent, wherein: hemolysis is H 2 O; latex concentration is 0.1%, suspended in the glycine buffer solution with a concentration of 15mmol / l; Antibody A is a goat anti-mouse IgG antibody with a concentration of 0.006mg / ml, mixed in a glycine buffer solution with a concentration of 60mmol / l; Antibody B is a mouse anti-human HbA1c monoclonal antibody with a concentration of 0.05 mg / ml, which is mixed in a glycine buffer with a concentration of 60 mmol / l; the amount of whole blood sample is 5 μl, the amount of hemolysis is 500 μl, and the amount of latex is 210 μl , the volumes of Antibody A and Antibody B were 50 μl and 70 μl, respectively. Wherein, the percentages of hemolysis, latex, goat anti-mouse IgG antibody and mouse anti-human HbA1c monoclonal antibody are 1.49%: 62.69%: 14.92%: 20.90%.

[0025] In this embodiment, if the concentration of the ...

Embodiment 2

[0027] In this embodiment, the method for directly measuring the percentage of glycosylated hemoglobin using the timing nephelometric method as the detection principle includes:

[0028] a. The making of the standard curve for the determination of glycated hemoglobin, that is, the corresponding database between the scattering value and the percentage of glycated hemoglobin-scattering value of multiple glycated hemoglobin standard products: the detection principle is based on the timing scattering turbidimetry, and the required materials include those described in the previous examples. glycosylated hemoglobin detection reagent, including 500 μl of hemolysis, 210 μl of latex, 50 μl of antibody A (goat anti-mouse IgG antibody with a concentration of 0.006 mg / ml), and 50 μl of antibody B (mouse anti-human HbA1c monoclonal antibody with a concentration of 0.05 mg / ml) Cloned antibody) 70μl; glycosylated hemoglobin standard and specific protein analyzer.

[0029] Sample processing: ...

Embodiment 3

[0037] Example of sensitivity verification of rapid detection reagents for glycated hemoglobin: Prepare rapid detection reagents for glycated hemoglobin, such as the reagents described in the previous examples, high-value and low-value quality controls, and a specific protein analyzer. Take a traceable high-value quality control substance and a low-value quality control substance, perform 10 tests on each quality control substance, and calculate the average value, standard deviation and variation coefficient of the test results. The results are shown in Table 1:

[0038] Table 1

[0039]

[0040]

[0041] From the coefficient of variation in Table 1, it can be seen that the C-reactive protein detection method provided by the present invention has relatively high precision.

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Abstract

The invention relates to a method for measuring glycosylated hemoglobin content, which comprises the following steps of: a, uniformly mixing a whole blood sample and hemolytic liquid; b, adding an appropriate amount of hemolyzed sample into latex suspending in glycine buffer solution; c, respectively adding an antibody B reagent and an antibody A reagent after preset time; d, uniformly mixing andreading a scattering value through a specific protein analyzer under the condition of single-wavelength light irradiation; and e, reading the glycosylated hemoglobin content from a standard curve diagram according to the scattering value. The glycosylated hemoglobin content of the blood sample can be directly determined without separately measuring the total hemoglobin content and the glycosylated hemoglobin content of the blood sample by taking fixed-time nephelometry as a detection principle; and antibody working solution does not need preparing in advance, so that the effective service time of the reagents is prolonged, the operating steps are simplified and the measuring time of the sample is shortened.

Description

technical field [0001] The invention relates to medical immunization, in particular to a method capable of directly measuring the percentage of glycosylated hemoglobin and its reagent. Background technique [0002] Glycated hemoglobin (glycosylated hemoglobin, HbA1c) is the product of the combination of hemoglobin and blood sugar in red blood cells in human blood. Glycosylated hemoglobin can stably and reliably reflect the average blood sugar level within 120 days before the test, and is not greatly affected by factors such as blood draw time, fasting, and insulin use. Therefore, the International Diabetes Federation has launched a new edition of the Asia-Pacific Diabetes Prevention Guidelines, which clearly stipulates that glycosylated hemoglobin is the internationally recognized "gold standard" for diabetes monitoring. If the fasting blood sugar or postprandial blood sugar is not well controlled, it is impossible for the glycosylated hemoglobin to reach the target. [00...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/72G01N21/82
Inventor 陈明峰余嘉陵杨青郑筱雯
Owner SHENZHEN GOLDSITE DIAGNOSTICS
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