The joint detection of glycated hemoglobin (HbA1c) indicators makes up for the shortcomings of simple blood glucose concentration detection in the diagnosis and treatment of diabetes and diabetic complications, and helps to detect diabetes and related complications early and take targeted treatment measures in a timely manner.With the application of more and more HbA1c detection technologies, the accuracy of HbA1c detection has been significantly improved, and the detection and analysis operation of this indicator has become more and more convenient.
1.Immunization method
(1).Immunoturbidimetry
The immunoturbidimetric method for detecting HbA1c levels is highly accurate and operable, and has been used in many diabetes research and clinical practices. After standardizing the serum specimen, mix the prepared HbA1c antibody buffer into the specimen to be tested, so that the anti-HbA1c antibody binds to HbA1c to generate an antigen-antibody complex, and then continue to add the polyhapten buffer to bind the residual anti-HbA1c in the reaction solution. The antibody produces an insoluble antigen-antibody complex, and the HbA1c level in the specimen is detected by turbidimetric method, and the Hb level is detected through another channel, and the HbA1c percentage level is calculated according to the formula. This detection method requires short time, low cost, repeatability and easy operation. However, it is easy to be cross-contaminated and affected by jaundice samples and lipid turbid samples. It is difficult to complete the test when the concentration is higher than the standard. Changes in Hb and HbF can affect the test results.
(2).Ion capture method
After the antibodies in the serum specimen combine with HbA1c, they can be processed by fluorescent labeling to generate complexes, which can then be combined with negatively charged polyanionic complexes and adsorbed on the surface of positively charged fibers. After cleaning and processing, the fluorescence intensity is detected and analyzed. Get the HbA1c level. This detection method has good specificity, high sensitivity, repeatability and recovery, is suitable for automated detection, and has minimal cross-contamination.
(3).Latex agglutination method
This method uses antigen-antibody agglutination reaction to process serum specimens, and directly detects the HbA1c and Hb levels contained in the processed serum specimens. This detection method takes a short time, has high specificity and accuracy, and is repeatable. The detection can be completed intelligently by a fully automatic biochemical instrument, saving labor costs and reducing error rates. However, this detection technology is not suitable for single specimen detection operations and is easily interfered by the levels of triacylglycerol, bilirubin, and glucose in the specimen.
2.Enzyme method
Hemolytic processing of blood samples, through specific endonuclease treatment, converts Hb enzyme into fructose amino acids, and generates hydrogen peroxide under the action of fructose amino acid oxidase. The HbA1c level in the blood sample is positively correlated with the hydrogen peroxide level test results, which promotes the Hydrogen oxide and catalase undergo chromogen coupling, observe the color change during the reaction, analyze the hydrogen peroxide level, and then infer the HbA1c level in the specimen.
3.Ion exchange chromatography
This detection method includes manual micro-column method and high-performance liquid chromatography. It can produce products with different charges through the glycation of the N-terminal valine of the hemoglobin β chain, creating a neutral pH environment. The number of positive charges carried by HbA1c is relatively small. It is treated by ion exchange chromatography to separate it from HbS, HbF, etc. The results obtained by this method are highly accurate and reproducible. The American Society of Clinical Chemistry recommends that the HbA1c results obtained by this technology be used as the gold standard for HbA1c testing and clinical diagnosis and treatment of diabetes and other diseases.
4.Electrophoresis method
The common electrophoresis method is agarose gel electrophoresis. The degree of Hb adsorption and charge on the gel in this method will directly affect the electrophoretic migration of Hb and agarose gel after treatment with acidic buffer.
5.Affinity chromatography
It utilizes the reversible binding reaction between the glucose cis-diol group contained in the Hb molecule and boric acid, uses m-aminophenylboronic acid agarose to participate in the reaction, and adds blood samples to the chromatography column to promote the binding reaction to occur in the column. Non-HbA1c components flow out of the chromatography column on their own, and then high-concentration polyhydroxy complexes that also contain cis-diol groups, such as sorbitol, are added to replace the combined products of HbA1c and boric acid. HbA1c can be calculated by measuring the content of the eluted products. value. This detection method is not sensitive to variant Hb and pathological Hb. The result of direct measurement is the total amount of HbA1c. The results obtained by this detection method will not be interfered by Hb variants and degradation products other than HbC and HbS. The results are relatively accurate and can be Cooperate with instant detection and analysis of individual specimens.
The HbA1c combined blood glucose value detection and analysis method is the foundation for the diagnosis and treatment of diabetes and related complications. Flexible selection of appropriate HbA1c detection technology according to needs is crucial to the accuracy of HbA1c value detection and the effect of comprehensive clinical diagnosis and treatment. In recent years, new technologies for HbA1c detection and analysis have continued to emerge, and more and more reference standards for HbA1c detection have emerged clinically. HbA1c clinical detection methods and judgment standards will gradually be unified. At present, domestic HbA1c detection research obviously lags behind foreign countries. Domestic clinical research should increase HbA1c detection and analysis research to facilitate diabetes diagnosis and treatment.
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