CN112067562A - Calibration product for fructosamine detection and detection kit using same - Google Patents
Calibration product for fructosamine detection and detection kit using same Download PDFInfo
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- CN112067562A CN112067562A CN202010768127.3A CN202010768127A CN112067562A CN 112067562 A CN112067562 A CN 112067562A CN 202010768127 A CN202010768127 A CN 202010768127A CN 112067562 A CN112067562 A CN 112067562A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/12—Circuits of general importance; Signal processing
- G01N2201/127—Calibration; base line adjustment; drift compensation
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Abstract
The invention discloses a calibrator for fructosamine detection and a detection kit using the calibrator. The invention also discloses a detection kit for fructosamine determination, and the detection kit adopts the bovine serum albumin as a calibrator. The method for determining the amine (FA) in the fruit pond by taking the bovine serum albumin as the calibrator (tertiary) is reasonably established on the basis of determining the amine (FA) in the fruit pond by using the nitrotetrazolium chloride (NBT) reduction method by taking 1-deoxy-1-morpholinofructose (DMF) as the primary standard substance. Meanwhile, the method has the advantages of accurate sample detection result, low cost and good transportation stability.
Description
Technical Field
The invention relates to the technical field of biological detection, in particular to a calibrator for fructosamine detection and a detection kit using the calibrator.
Background
Fructosamine is also called glycated serum protein, is a substance formed by plasma protein in the process of glucose non-enzymatic glycation, and can reflect the average blood glucose level of a diabetic patient within 1-3 weeks before detection due to the half-life of the plasma protein of 17 days. Makes up for the deficiency that the glycosylated hemoglobin can not reflect the change of the blood sugar concentration in a short period to a certain extent. Fructosamine is a rapid and inexpensive assay (chemical method) and a good index for the evaluation of diabetes control.
Since the establishment of nitrotetrazolium chloride (NBT) reduction method for Fructosamine (FA) by Johnson in 1982, it has been widely used for monitoring blood sugar control of diabetic patients at home and abroad. The most commonly used standard for this method is 1-DEOXY-1-MORPHOLINO-D-FRUCTOSE (1-DEOXY-1-MORPHOLINO-D-FRUCTOSE, DMF) or the reaction of Bovine Serum Albumin (BSA) with glucose to produce fructosamine.
In the above detection method, the standard substance is DMF, and DMF is a small molecule substance and cannot represent the true structure of human serum fructosamine (i.e. glycated serum protein is a large molecule substance). Glycated serum proteins are difficult to extract, and fructosamine is artificially synthesized from bovine serum albumin and glucose, and since this non-enzymatic reaction is a slow process, its detection value changes with time, and thus it is not suitable for use as a calibrator.
Disclosure of Invention
Therefore, the invention provides a calibrator for fructosamine detection and a detection kit using the calibrator.
In order to achieve the above purpose, the invention provides the following technical scheme:
a calibrator for fructosamine detection, said calibrator being bovine serum albumin.
In one embodiment of the present invention, the concentration of bovine serum albumin is 50g/L, which corresponds to a fructosamine concentration assignment of 285. mu. mol/L.
In one embodiment of the invention, the fructosamine detection is performed by nitrotetrazolium chloride blue reduction.
In one embodiment of the invention, the bovine serum albumin is calibrated under conditions of 37 ℃ temperature, 10.35 pH and 0.5mmol/L NBT concentration.
The present invention also provides a detection kit for fructosamine assay, which uses the bovine serum albumin of claim 1 as a calibrator.
In one embodiment of the invention, the detection kit further comprises a full-automatic biochemical analyzer.
In one embodiment of the present invention, when the fully automatic biochemical analyzer detects fructosamine, the determination conditions are as follows: the reaction temperature is 37 ℃, the main wavelength is 546nm, and the auxiliary wavelength is 700 nm.
In one embodiment of the invention, in the test kit, the concentration of fructosamine in the serum sample (assay tube absorbance value/calibrator tube absorbance value) is the assigned concentration of fructosamine for the calibrator.
The invention has the following advantages:
the method for determining the amine (FA) in the fruit pond by taking the bovine serum albumin as the calibrator (tertiary) is reasonably established on the basis of determining the amine (FA) in the fruit pond by using the nitrotetrazolium chloride (NBT) reduction method by taking 1-deoxy-1-morpholinofructose (DMF) as the primary standard substance.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
The structures, ratios, sizes, and the like shown in the present specification are only used for matching with the contents disclosed in the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions that the present invention can be implemented, so that the present invention has no technical significance, and any structural modifications, changes in the ratio relationship, or adjustments of the sizes, without affecting the effects and the achievable by the present invention, should still fall within the range that the technical contents disclosed in the present invention can cover.
FIG. 1 is a graph showing the correlation between reagents for glycated serum proteins (NBT method) which were calibrated with 1-deoxy-1-morpholinofructose (DMF) calibrator and Bovine Serum Albumin (BSA) calibrator, respectively, according to an embodiment of the present invention.
Detailed Description
The present invention is described in terms of particular embodiments, other advantages and features of the invention will become apparent to those skilled in the art from the following disclosure, and it is to be understood that the described embodiments are merely exemplary of the invention and that it is not intended to limit the invention to the particular embodiments disclosed. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1 calibrator and kit for fructosamine detection
The embodiment of the invention provides a novel calibration product bovine serum albumin for fructosamine detection. Specifically, in this example, on the basis of measuring Fructosamine (FA) by nitrotetrazolium chloride (NBT) reduction method using 1-deoxy-1-morpholinofructose (DMF) as primary standard, it is reasonably established to measure Fructosamine (FA) using bovine serum albumin as secondary standard, and experimental observation and analysis are performed on the performance of this secondary standard. Wherein the name of 1-DEOXY-1-morpholinofructose is DEOXY-1-MORPHOLINO-D-FRUCTOSE, which is abbreviated as DMF, and the chemical structure and the reaction with NBT are similar to fructosamine.
In the embodiment of the invention, a calibrator (third-level) prepared by taking bovine serum albumin as a main raw material is used for determining the concentration of Fructosamine (FA) in a sample: the measurement is carried out by a single reagent end-point method using a fully automatic biochemical analyzer such as Hitachi 7180 fully automatic analyzer. Placing a glycosylated serum protein (NBT method) reagent on a corresponding reagent position, placing distilled water and a calibrator on a corresponding position of a calibration plate, placing a sample to be tested on a sample plate, and calibrating by using bovine serum albumin of the embodiment of the invention, wherein the reaction conditions are that the temperature is 37 ℃, the pH value is 10.35, and the reaction condition is 0.5mmol/L nitrotetrazolium chloride (NBT). The specific operation is shown in the determination conditions of Table 1, and the specific operation steps are shown in Table 2.
TABLE 1
TABLE 2
And (3) adopting a one-point linear calibration to make a standard curve according to the concentration of the calibrator and the corresponding absorbance, wherein the concentration of Fructosamine (FA) in the sample is calculated from the standard curve through the absorbance of the sample.
The Fructosamine (FA) concentration in the sample was calculated by the formula: serum sample Fructosamine (FA) concentration (assay tube absorbance/calibrator tube absorbance) bovine serum albumin calibrant Fructosamine (FA) assigned concentration.
In this example, the correlation coefficient (r) is not less than 0.95 when measured values of Fructosamine (FA) measured using DMF at a concentration of 0.28mmol/L as a calibrator and bovine serum albumin at a concentration of 50g/L as a calibrator are compared. The calibrator prepared from bovine serum albumin with the concentration of 50g/L is characterized in that the measured value of the calibrator is compared with the measured value of 0 day after the calibrator is accelerated for 3 days at 42 ℃, and the variation of the measured value is within +/-10%.
Example 2 calibration of 1-deoxy-1-Morpholinulose (DMF) calibrators bovine serum Albumin calibrators of the invention
The accuracy test of sample detection is carried out on bovine serum albumin (third level) which is a calibrator prepared by the invention and a first level standard substance prepared by 1-deoxy-1-morpholine fructose (DMF).
1. Experimental materials: calibrator-1: 1-deoxy-1-morpholin-D-fructose (0.28mmol/L), proclin-300(0.03ml/L), assigned concentrations: 280 mu mol/L; calibrator-2: BSA (50g/L), proclin-300(0.03ml/L), assigned concentrations: 285 mu mol/L; glycated serum protein (NBT method) assay kit (Haifeng Biotechnology (Bio) Ltd.).
2. The experimental method comprises the following steps: the calibrator-1 and the calibrator-2 are respectively used as the calibration of a glycosylated serum protein (NBT method) determination kit, and 100 serum samples (abnormal samples are not less than 10%) are detected.
3. As a result of the measurement, the results of the detection of calibrator-1 and calibrator-2 are shown in Table 3.
TABLE 3
As shown in fig. 1, the calibration formula is that y is 0.996x +1.1, and the correlation coefficient (r) is 0.99, which is greater than 0.975, and is acceptable.
3. And (4) conclusion: from the above results, it can be seen that Fructosamine (FA) measured with calibrator-2 containing bovine serum albumin as the main component in the examples of the present invention has a higher degree of agreement in the sample detection results than FA measured with calibrator-1 containing 1-deoxy-1-morpholine-D-fructose as the main component.
Example 3 transport stability test of bovine serum albumin calibrator according to the present invention
1. Experimental materials: calibrator-2: BSA (50g/L), proclin-300(0.03 ml/L); glycated serum protein (NBT method) assay kit (Haifeng Biotechnology (Bio) Ltd.).
2. The experimental method comprises the following steps: placing 1 calibrator-2 into 42 deg.C incubator, taking out on day 3, and detecting with calibrator-2 on day 0.
3. As a result of the experimental measurement, the accelerated stability of calibrator-2 was as shown in Table 4.
TABLE 4
4. And (4) conclusion: the test results show that the transportation stability of the calibrator-2 prepared by taking bovine serum albumin as the main component is qualified after 3 days of acceleration at 42 ℃.
The bovine serum albumin calibrator is qualified in transportation stability at 42 ℃ and accurate in sample detection in Fructosamine (FA) detection determined by a nitrotetrazolium chloride (NBT) reduction method, can ensure the accuracy of Fructosamine (FA) detection determined by a clinical nitrotetrazolium chloride (NBT) reduction method, is relatively low in cost, and is a substitute for the calibrator component 1-DEOXY-1-MORPHOLINO-D-FRUCTOSE (1-DEOXY-1-MORPHINO-D-FRUCTOSE, DMF) in Fructosamine (FA) detection determined by the nitrotetrazolium chloride (NBT) reduction method.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (8)
1. A calibrator for fructosamine detection, wherein the calibrator is bovine serum albumin.
2. The calibrator for fructosamine detection according to claim 1,
the concentration of the bovine serum albumin is 50g/L, and the corresponding fructosamine concentration assignment is 285 mu mol/L.
3. The calibrator for fructosamine detection according to claim 1,
the fructosamine detection adopts a nitrotetrazolium chloride blue reduction method.
4. The calibrator for fructosamine detection according to claim 1,
the calibration reaction conditions of the bovine serum albumin are that the temperature is 37 ℃, the pH is 10.35, and the concentration of NBT is 0.5 mmol/L.
5. A detection kit for fructosamine assay, characterized in that, said detection kit adopts bovine serum albumin of claim 1 as calibrator.
6. The detection kit for fructosamine assay according to claim 5, characterized in that,
the detection kit also comprises a full-automatic biochemical analyzer.
7. The detection kit for fructosamine assay according to claim 6, characterized in that,
when the full-automatic biochemical analyzer detects fructosamine, the determination conditions are as follows: the reaction temperature is 37 ℃, the main wavelength is 546nm, and the auxiliary wavelength is 700 nm.
8. The detection kit for fructosamine assay according to claim 6, characterized in that,
in the detection kit, the fructosamine concentration in a serum sample is (determination tube absorbance value/calibrator tube absorbance value) and the calibrator fructosamine assigned concentration.
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| CN202010768127.3A CN112067562A (en) | 2020-08-03 | 2020-08-03 | Calibration product for fructosamine detection and detection kit using same |
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| CN202010768127.3A CN112067562A (en) | 2020-08-03 | 2020-08-03 | Calibration product for fructosamine detection and detection kit using same |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112326639A (en) * | 2020-11-25 | 2021-02-05 | 迈克生物股份有限公司 | Kit and method for detecting fructosamine |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112326639A (en) * | 2020-11-25 | 2021-02-05 | 迈克生物股份有限公司 | Kit and method for detecting fructosamine |
| CN112326639B (en) * | 2020-11-25 | 2024-01-05 | 迈克生物股份有限公司 | Kit and method for detecting fructosamine |
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