CN109470533B - Preparation method of human whole blood matrix quality control product for portable glucometer - Google Patents

Abstract

The invention discloses a preparation method of a human-derived whole blood matrix quality control product for a portable blood glucose meter, which includes the steps of separating whole blood cells, fixing whole blood cells, and preparing quality control products with three blood sugar concentrations of high, medium and low. The present invention can timely and effectively detect possible errors in the detection of the high, medium and low blood glucose concentrations of the medical blood glucose determination value level by the portable blood glucose meter by using the quality control products with the high, medium and low blood glucose concentration levels. . The quality control product prepared by the method of the invention is fixed with human whole blood cells, which ensures that the quality control product is a whole blood matrix and is consistent with the type of the sample identified by the blood glucose meter; and also solves the problem of glycolysis of blood sugar in the blood. , stabilized blood sugar levels. The quality control product prepared by the method of the invention has good interoperability and can be applied to various portable blood glucose meters of different brands and models.

Description

A kind of preparation method of human whole blood matrix quality control substance for portable blood glucose meter

technical field

The invention relates to the technical field of standard products used in clinical testing processes, in particular to a preparation method of a human-derived whole blood matrix quality control product for portable blood glucose meters, in particular to the preparation of a variety of blood glucose products suitable for different brands or models The method for the human-derived whole blood matrix quality control of the instrument.

Background technique

Nowadays, POCT (point-of-care testing, point-of-care testing) technology is more and more favored by clinicians due to its advantages of point-of-care testing, such as rapidity, simplicity, small sample size, and potential improvement of patient prognosis. As the most frequently used POCT instrument in medical institutions, the accuracy and reliability of portable blood glucose meters play a crucial role in the monitoring and management of blood glucose of patients in medical institutions.

However, due to the lack of effective reference materials (reference materials, RMs) for quality control of portable blood glucose meters, the traceability and measurement accuracy of the detection method cannot be fully guaranteed, so the effective value cannot be transmitted, which often leads to the difference between the test results and the center. There are large differences between laboratories, and some blood glucose meters have been in a substandard state for a long time and are not easy to be found, thus affecting the clinical implementation of reasonable and effective diagnosis and treatment measures.

In order to standardize the detection quality of portable blood glucose meters in medical institutions and improve the application value of portable blood glucose meters in the diagnosis and treatment of diabetes, selecting appropriate reference materials for quality control is one of the keys to exerting its clinical application value. As a kind of reference material, quality control materials (QCMs) are mainly used to monitor the performance of a confirmed experimental method, and can detect the change of the experimental method over time or the occurrence of statistical out-of-control conditions. External quality assessment/proficiency testing (external quality assessment, EQA/proficiency testing, PT) or internal quality control (internal quality control, QC). Among them, inter-laboratory quality assessment/proficiency testing means that multiple specimens are periodically sent to laboratories for analysis and/or identification, and the results of each laboratory are compared with the results of other laboratories in the same group or specified values. , and report the results of the comparison to the participating laboratories. Indoor quality control refers to the use of certain methods and steps to continuously evaluate the reliability of laboratory work, aiming to monitor the precision of the laboratory's routine work and improve the consistency of intra- and inter-batch sample testing in the laboratory's routine work. , an effort to determine whether laboratory results are reliable and reportable.

At present, most of the blood glucose calibrators or quality control products used to verify the precision, stability and accuracy of blood glucose meters are non-human whole blood substrates, while portable blood glucose meters are mainly used to detect glucose in human whole blood. The test results obtained from the source whole blood matrix as the quality control material will be different from the actual results; secondly, different brands of blood glucose meters have different quality control materials, and even different types of blood glucose meters of the same brand have different quality control materials. The same quality control product is measured on different brands of instruments, the results will be different, and the target value of the same quality control product measured on different models of instruments will also be different, indicating that the test results of the quality control product cannot be communicated. Not only that, the supporting calibrators or quality control products provided by manufacturers are expensive, and some imported reagents are also restricted by transportation conditions and supply chains. This not only increases medical costs, but also creates hidden dangers for the management and treatment of diabetic patients.

Interoperability (Commutability) is an important attribute of a reference material, which refers to the numerical relationship between the measurement results obtained by each measurement procedure when the substance is measured by different measurement procedures, and the numerical relationship between the measurement results obtained when using these measurement procedures to measure actual clinical samples. the consistency of the relationship. Based on the above defects, it can be seen that the development of an interoperable human-derived whole blood matrix quality control product for portable blood glucose meters is a substantial breakthrough in improving the blood sugar management of diabetic patients. The preparation method of such a human-derived whole blood matrix quality control product is also essential areas that need to be addressed urgently.

SUMMARY OF THE INVENTION

The purpose of the present invention is to aim at the technical defects existing in the prior art, to provide a preparation method of a human-derived whole blood matrix quality control product for a portable blood glucose meter, and the prepared human-derived whole blood matrix quality control product can be used for different Portable blood glucose meters of different brands, and the test results are interoperable between the blood glucose meters, including the separation of whole blood cells from human whole blood, the fixation of whole blood cells to obtain fixatives, and the preparation of quality control products with three blood glucose concentrations of high, medium and low, etc. step.

The human whole blood matrix quality control product is of human origin, and is prepared from the following raw materials: human whole blood, fixative, and physiological saline.

The fixative is selected from solutions containing formaldehyde and/or glutaraldehyde.

The fixative is a PBS solution containing 2-8% (v/v) formaldehyde and 2-8% (v/v) glutaraldehyde; preferably, the fixative is a PBS solution containing 2-6% (v/v) ) formaldehyde and 2-6% (v/v) glutaraldehyde in PBS; preferably 4% (v/v) formaldehyde and 4% (v/v) glutaraldehyde in PBS.

The high blood sugar concentration is 12.0-16.0 mmol/L, the middle blood sugar concentration is 6.0-8.0 mmol/L, and the low blood sugar concentration is 2.0-3.0 mmol/L in the quality control product of the three blood sugar concentrations of high, middle and low.

The human whole blood is venous blood containing red blood cells, white blood cells and platelets of normal people aged 18-50.

The described separation of whole blood cells is as follows: human whole blood is centrifuged at 1500 g for 10 min, the upper layer is discarded, and the lower layer is the whole blood cells.

The fixation obtained by fixing the whole blood cells is specifically as follows: adding the whole blood cells to the fixative for fixing at room temperature for 24-48 hours, and the volume ratio of the whole blood cells to the fixative is 1:(5-10);

It also includes the operation of purifying the fixation, specifically: centrifuging the whole blood cells after the fixation is completed, discarding the upper fixative, adding physiological saline to the lower fixation to wash 3-5 times, and the volume ratio of the fixation to the physiological saline is 1:( 5-10), and then filtered, the filtrate is the purified immobilizer.

Described preparing the quality control substance of three blood sugar concentrations of high, middle and low is specifically: adding pre-prepared plasma mixtures of different blood sugar concentrations to the fixative or the purified fixative respectively, and the volume ratio of the fixation to the plasma is 1:(1- 2) (preferably 1:1), respectively obtain the low blood sugar concentration quality control product, the medium blood sugar concentration quality control product, and the high blood sugar concentration quality control product; Centrifuge, take the supernatant, or add glucose after taking the supernatant.

The invention provides a preparation method of a human-derived whole blood matrix quality control substance suitable for different brands and different models of portable blood glucose meters. Quality controls prepared by this method:

First, three concentrations of high, medium, and low levels of medical blood glucose determination values are involved. The normal blood sugar level is between 4.4-6.0mmol/L. Too low or too high blood sugar will cause harm to the human body. In medical institutions, the blood glucose level of patients is mainly detected by portable blood glucose meters, and the dose of insulin therapy is adjusted. If the blood sugar result detected by the portable blood glucose meter is too high or too low, it will directly affect the dosage of insulin, increase the medical risk, and seriously lead to hypoglycemia coma or ketoacidosis caused by hyperglycemia. The present invention can timely and effectively detect possible errors in the detection of the high, medium and low blood glucose concentrations of the medical blood glucose determination value level by the portable blood glucose meter by using the quality control products with the high, medium and low blood glucose concentration levels. .

Secondly, the quality control product prepared by the method of the present invention is fixed with human whole blood cells, on the one hand, it ensures that the quality control product is a whole blood matrix, which is consistent with the type of the sample identified by the blood glucose meter; and also solves the problem that the glycolysis of blood sugar will occur in the blood. problem, stabilize blood sugar values.

Thirdly, it is proved by experiments that the quality control product prepared by the method of the present invention has good interoperability and can be applied to various portable blood glucose meters of different brands and models.

Finally, the quality control product prepared by the method of the invention has the advantages of wide raw material sources, simple preparation process and low cost, which can meet the clinical requirements for the quality control of the portable blood glucose meter, and has great market promotion and application value.

Description of drawings

Fig. 1 shows the schematic flow sheet of the preparation method of the present invention;

Figures 2-4 are graphs showing the effect of evaluating the interoperability of the quality control products prepared by the method of the present invention on blood glucose meters of different brands.

Detailed ways

At present, the quality control materials for blood glucose detection at home and abroad are mostly serum matrix or water matrix, and there is a lack of whole blood matrix quality control materials suitable for portable blood glucose meters. Blood sugar is prone to glucolysis (the blood sugar value in normal people's whole blood will undergo glycolysis at a rate of 5-7%/hour), which in turn causes the blood sugar value to decrease. Although synthetic latex particle quality control materials or serum matrix quality control materials are used for the quality control of portable blood glucose meters at home and abroad, the obtained quality control materials are still unable to be suitable for various types of blood glucose meters of different brands. Based on the above reasons, a whole blood matrix quality control product is developed to make the test results of different brands and different models of blood glucose meters interoperable, and applied to the quality control of portable blood glucose meters in medical institutions, which can effectively reduce medical costs and improve Application value of portable blood glucose meter in blood glucose monitoring.

The invention provides a preparation method of a human whole blood matrix quality control product. The quality control product prepared by the method is of human origin and is prepared from the following raw materials: the raw materials include normal whole blood, fixative, physiological saline and the like. in,

Normal whole blood is the venous blood of healthy people aged 18-50, anticoagulated with EDTK-K ₂ , the same blood type, including red blood cells, white blood cells and platelets;

The fixative can be selected from formaldehyde and/or glutaraldehyde. Formaldehyde has a small molecular weight, strong penetrating ability and mild reaction. It can be used for pre-fixation of cell histochemical studies, but it has poor preservation of cell matrix and most of the matrix is lost after dehydration. ; and glutaraldehyde as a fixative, the reaction speed is fast, but the penetration speed is slower than that of formaldehyde, and it has the best effect when used in conjunction with formaldehyde. The fixative preferably contains 2-8% (v/v) formaldehyde and 2-8% (v/v) glutaraldehyde in PBS solution (PBS buffer is dissolved in 1 L of distilled water by 10.2 g of phosphate, and the pH is adjusted with dilute hydrochloric acid. to 7.2), more preferably 2-6 (v/v) % formaldehyde and 2-6 (v/v) % glutaraldehyde in PBS, most preferably 4 % (v/v) formaldehyde and 4 % (v/v) glutaraldehyde in PBS.

The method for preparing the human whole blood matrix quality control product provided by the present invention specifically includes the following steps:

(1) Take normal whole blood, centrifuge at 1500g for 10min in a low-speed centrifuge, the upper layer is plasma, the lower layer is whole blood cells, and the plasma and whole blood cells are separated for use;

(2) adding the whole blood cells obtained in step (1) into a fixative, and fixing at room temperature for 24-48h; the volume ratio of the whole blood cells to the fixative is 1:(5-10);

(3) Centrifuge at 1500g for 10 min after fixation, discard the upper fixative, add 0.9% normal saline to wash the lower fixative for 3-5 times, the volume ratio of fixation to normal saline is 1:(5-10), wash The specific steps are as follows: fully mix the fixation with physiological saline, centrifuge at 1500g for 10 min, discard the upper washing solution, and leave the fixation; then filter the washed lower fixation with a filter screen (pore size is 40 μm) to remove precipitates and impurities, The filtrate is the purified fixative;

(4) Plasma with different blood glucose concentrations is added to the purified fixative obtained in step (3), and placed on a shaker for mixing for 30 minutes to obtain a quality control product with low blood sugar concentration, a quality control product with medium blood sugar concentration, and a quality control product with high blood sugar concentration. The volume ratio of fixed substance and plasma is 1:(1-2) (preferably 1:1); plasma with different blood glucose concentrations is human plasma, because the blood glucose measurement range of most portable blood glucose meters is between 1.1-33.3mmol/L Therefore, in the present invention, the quality control product is set to three blood sugar levels: low, medium and high, and the blood sugar level of the quality control product is achieved by plasma with different blood sugar concentrations. The preparation process of plasma with different blood sugar concentrations is as follows: (1) hypoglycemia Concentration plasma: select venous whole blood with a blood sugar concentration of about 4.0-6.0 mmol/L, centrifuge at 1500g for 10 min, take the supernatant, and obtain the low blood sugar concentration plasma, which is the plasma required for the preparation of the low blood sugar concentration quality control product; (2) Medium Blood glucose concentration plasma: select venous whole blood with a blood glucose concentration of about 4.0-6.0 mmol/L, centrifuge at 1500g for 10 minutes, take the supernatant, and then add 40 μl of 200 mmol/L glucose solution per milliliter of supernatant, which is medium blood glucose concentration plasma, that is, preparation (3) Plasma with high blood sugar concentration: select venous whole blood with a blood sugar concentration of about 4.0-6.0 mmol/L, centrifuge at 1500g for 10 min, take the supernatant, and then add 200 mmol/L per ml of supernatant. L glucose solution 100μl, high blood sugar concentration plasma, that is, the plasma required for the preparation of high blood sugar concentration quality control products;

(5) The quality control products with low, medium and high blood glucose concentration obtained in step (4) are divided into sterile small test tubes, stored at 2-8°C, and fully mixed before each use.

In order to prolong the shelf life of the quality control product of the present invention, the purified fixative obtained in step (3) can also be added to 0.9% normal saline to suspend, and the volume ratio of the fixative to normal saline is 1: (1-2), divided into It was placed in a sterile small test tube, and then centrifuged at 1500 g for 10 min in a low-speed centrifuge. The upper layer was physiological saline, and the lower layer was purified fixative, which was stored at 2-8°C. The plasma of different blood glucose concentrations obtained in step (4) was divided into sterile small test tubes and stored at -20°C for a long time. That is, the purified fixative is packaged separately from plasma with different blood glucose concentrations. When using, the upper layer of physiological saline on the liquid-sealed fixative is discarded, and the plasma with different blood glucose concentrations is reconstituted at 2-8 °C, and then the fixative is mixed with plasma. The quality control products of the present invention with low, medium and high blood glucose concentrations are obtained by the existing preparation.

The quality control product of the present invention does not need to distinguish blood types, only normal whole blood of the same blood type is mixed and centrifuged in step (1), and the quality control product of the present invention can be used directly without distinguishing blood types.

The present invention focuses on reducing the activity of the cells by fixing the cells with aldehydes, thereby inhibiting the glycolysis in the whole blood cells and maintaining the stability of the blood sugar value. On the one hand, the presence of blood cells in the quality control product prepared by the method of the present invention satisfies the requirement that the portable blood glucose meter needs to reach 20-60% hematocrit (HCT). On the other hand, this quality control product does not add any other substances (such as polysaccharides such as sucrose, glucose, fructose, galactose, lactose, maltose, starch and dextrin, antibiotics and sodium citrate, etc.) Preservatives) to ensure the consistency with the patient sample matrix and avoid the interference of various substances during the detection process of the portable blood glucose meter, such as maltose, xylose, galactose, etc. Unstable results. In addition, the quality control product prepared by the method of the present invention uses human blood instead of animal blood, mainly because the cell morphology of animal blood and human blood is quite different, and the hematocrit (HCT) of the two kinds of blood is quite different, which will also affect the detection. The accuracy of the results and the poor interoperability of quality control products between different brands of portable blood glucose meters. The quality control product prepared by the method of the invention satisfies the consistency between the matrix and the patient sample to the greatest extent, and does not add any interfering substances that affect the detection of the instrument, and at the same time maintains the stability of the blood glucose value, and can realize the detection of the quality control product only. It can judge the accuracy of the test results of the portable blood glucose meter, whether it needs to be corrected by the system, and whether the test results of the patient are acceptable.

The content of the present invention is described in more detail below with reference to specific embodiments, and the present invention is further elaborated, but these embodiments are by no means to limit the present invention.

Experimental example 1. The homogeneity verification of the human-derived whole blood matrix quality control product for the portable blood glucose meter prepared by the method of the present invention

With reference to the CNAS-GL29 standard issued by the China National Accreditation Service for Conformity Assessment - "General Principles and Statistical Methods for the Determination of Reference Materials/Standard Samples" on the evaluation of homogeneity research, use the HORIBA glucose electrode analyzer (LP-150C ) were tested, and 10 samples (numbered 1-10) were randomly selected from the quality control products of each blood glucose concentration (low, medium, and high) as samples for homogeneity evaluation, and each sample was repeated 3 times. The sequence of the first determination during the measurement process is: 1-3-5-7-9-2-4-6-8-10; the sequence of the second determination is: 10-9-8-7-6-5 -4-3-2-1; the order of the third determination is: 2-4-6-8-10-1-3-5-7-9, the results are shown in Table 1. Statistical one-way ANOVA was used for analysis, and the results are shown in Table 2.

To check the uniformity of the samples, i samples (i=1, 2, . . . m) were drawn, and each sample was tested j times (j=1, 2, . . . , n) under repeated conditions.

Test average for each sample

Overall average of all sample tests

total number of tests

均方MS₁＝SS₁/f₁ between-sample sum of squares

Mean square MS ₁ =SS ₁ /f ₁

均方MS₂＝SS₂/f₂ In-sample sum of squares

Mean square MS ₂ =SS ₂ /f ₂

Degree of freedom f ₁ =m-1

f ₂ =Nm

Statistics F = MS ₁ /MS ₂

If F < degrees of freedom (f ₁ , f ₂ ) and a critical value Fα(f ₁ , f ₂ ) for a given significance level α (usually α=0.05), it indicates that there is no significant difference within and between samples, The sample is homogeneous.

Table 1 Measurement data of the homogeneity study of the quality control substance (unit: mmol/L)

Table 2 Analysis of variance table for quality control homogeneity study

From the results in Table 1 and Table 2, it can be seen that the human whole blood matrix quality control product prepared by the method of the present invention for the portable blood glucose meter has no significant difference whether within the sample or between samples, indicating that the method of the present invention is prepared by the method of the present invention. The resulting control is homogeneous and can be used as a control for portable blood glucose meters.

Experimental example 2. Stability verification of the human whole blood matrix quality control product prepared by the method of the present invention

With reference to the CNAS-GL29 standard issued by the China National Accreditation Service for Conformity Assessment - "General Principles and Statistical Methods for the Determination of Reference Materials/Standard Samples", the stability evaluation requirements are used for the human whole blood matrix quality control product prepared by the method of the present invention. A short-term 2-8°C stability evaluation was performed. Three samples were randomly selected for each quality control of blood glucose concentration (low, medium, and high), and each sample was measured twice, and each measurement was completed within 10 minutes. The HORIBA glucose electrode analyzer (LP-150C) was used for continuous measurement for 20 days, and the differences between the results on the 2nd, 3rd, 5th, 10th, 15th, and 20th days and the results on the 1st day were observed. After the measurement, statistical t-test was used for analysis, and the results are shown in Table 3.

Calculate the t-value as follows:

—第一次检验测量数据的平均值；where:

- the average value of the measured data for the first inspection;

—第二次检验测量数据的平均值；

- the average value of the measurement data for the second inspection;

s ₁ - the standard deviation of the first inspection measurement data;

s ₂ - the standard deviation of the measurement data for the second inspection;

n ₁ - the number of measurements for the first inspection measurement;

n ₂ — The number of measurements for the second inspection measurement.

Note: In order to ensure the accuracy of the mean and standard deviation, both n1 and n2 are ≥6.

If t<significance level α (usually α=0.05) the degree of freedom is the critical value t _α(n1+n2-2) of n1+n2-2, then there is no significant difference between the two means.

Table 3 Measurement data of quality control product stability study (unit: mmol/L)

The results in Table 3 show that the results of the quality control products with high, medium and low blood sugar concentrations on the 15th day of storage are still no significant difference (P>0.05) compared with the first day (P>0.05), indicating that the quality control products prepared by the method of the present invention Control product stability is good.

Experimental Example 3. Interoperability Verification of the Human-derived Whole Blood Matrix Quality Control Product Prepared by the Method of the Invention for Portable Blood Glucose Meters

Collect 33 clinical specimens with blood glucose concentrations in the high, medium and low concentration ranges, respectively, and use different brands and models of blood glucose meters and large-scale biochemical analysis of the samples and the three quality control substances of blood glucose concentration to be verified. The 95% confidence interval of the blood glucose meter test for the central laboratory detection method is drawn, and it is judged whether the measured value of the quality control product falls within the 95% confidence interval, and the quality control product The measured value falls within the 95% confidence interval, indicating good interoperability, and if it does not fall within the 95% confidence interval, indicating poor interoperability, the results are shown in Figure 2-4 (in the figure: ● represents clinical samples, and ■ represents three concentrations Glucose controls, …… represent 95% confidence intervals).

This experiment involves 15 portable blood glucose meters, namely Roche ACCU-CHEK Performa, Roche ACCU-CHEK Active, Bayer CONTOUR TS, Bayer CONTOUR PLUS, Bayer Bayer1455, Abbott FreeStyleOptium, ARKRAY GT-1820, ARKRAY GT-1970, Terumo MEDISAFEFIT, HORIBA LP-150C, Johnson & Johnson StatStrip, Johnson & Johnson ONETOUCH UltraVue, Ritter Rightest GM300, Wanfu EC-102 and Botangping II.

The results in Figures 2-4 illustrate that the quality control products prepared by the method of the present invention are used in Roche ACCU-CHEK Performa, Roche ACCU-CHEK Active, Bayer CONTOUR TS, Bayer CONTOUR PLUS, Abbott FreeStyleOptium, ARKRAY GT-1820, Tyre The 9 portable blood glucose meters of Mao MEDISAFE FIT, HORIBA LP-150C and Johnson & Johnson StatStrip have good interoperability. Compared with the existing quality control product which can only be applied to a certain brand of portable blood glucose meter, the quality control product prepared by the method of the present invention is a breakthrough.

Experimental example 4. Indoor quality control application

Three quality control materials with high, medium, and low blood glucose concentrations were used for indoor quality control, and two quality control materials with two blood glucose concentrations matched with two portable blood glucose meters were selected as controls, and the detection was carried out continuously for 5 days, repeated 4 times a day. Portable Blood Glucose Meter 1: HORIBA Glucose Electrode Analyzer (LP-150C), the quality control products are QC1, QC2; Portable blood glucose meter 2: ROCHE, ACCU-CHEK Performa, the quality control products are QC3, QC4 respectively. Calculate the mean and standard deviation (SD) and coefficient of variation (CV) of the test results, and cooperate with the State Food and Drug Administration of the People's Republic of China to issue the national standard "General technical conditions for blood glucose monitoring systems for self-testing of in vitro diagnostic test systems [S/OL] ]》When the blood glucose concentration is less than 5.5mmol/L, the SD is less than 0.42mmol/L; when the blood glucose concentration is ≥5.5mmol/L, the CV is less than 7.5%.

Table 4 Indoor quality control application results of quality control products

quality control Mean (mmol/L) SD CV(%) high blood sugar concentration 14.48 0.09 0.63 blood sugar concentration 7.56 0.05 0.66 low blood sugar concentration 2.48 0.04 1.65 QC1 2.55 0.09 3.71 QC2 26.69 0.30 1.12 QC3 2.56 0.07 2.69 QC4 17.1 0.22 1.29

Table 4 shows that the SD value of the low blood sugar concentration quality control product prepared by the method of the present invention is 0.04, which is less than 0.42 mmol/L, and the CV values of the medium blood sugar concentration quality control product and the high blood sugar concentration quality control product are 0.66% and 0.63%, respectively. It is less than 7.5%, indicating that the three quality control products of blood glucose concentration meet the national standards, and the results are similar to those of commercial quality control products, indicating that they can be applied to the daily indoor quality control of portable blood glucose meters.

Quality control products can be divided into fixed-value quality control products and non-valued quality control products. The mean and standard deviation of non-determined quality control products are obtained by using routine laboratory methods to test at least 20 bottles in different days, or at least 4 repeated tests per day for at least 5 days. Since instruments suitable for whole blood matrix blood glucose detection, traceability, and quality control product assignment are not widely used in clinical medical institutions, the quality control product prepared by the method of the present invention is used as a non-determined value quality control product.

The above are only the preferred embodiments of the present invention. It should be noted that, for those skilled in the art, without departing from the principles of the present invention, several improvements and modifications can be made. These improvements and Retouching should also be considered as part of the present invention.

Claims (14)

1. A preparation method of a human whole blood matrix quality control product for a portable glucometer is characterized by comprising the following steps: separating whole blood cells from normal human whole blood, fixing the whole blood cells by using a fixing agent to obtain a fixed object, and preparing quality control products of high, medium and low blood glucose concentrations; the fixing agent is PBS solution containing 2-8% (v/v) formaldehyde and 2-8% (v/v) glutaraldehyde;

the quality control products for preparing the blood sugar concentrations of the high, middle and low blood sugar levels are specifically as follows: adding pre-prepared blood plasma with different blood sugar concentrations into a fixed object respectively, and mixing to obtain a low blood sugar concentration quality control product, a medium blood sugar concentration quality control product and a high blood sugar concentration quality control product respectively; the blood sugar concentration of the low blood sugar concentration quality control product is 2.0-3.0mmol/L, the blood sugar concentration of the medium blood sugar concentration quality control product is 6.0-8.0mmol/L, and the blood sugar concentration of the high blood sugar concentration quality control product is 12.0-16.0 mmol/L.

2. The preparation method of claim 1, wherein the human whole blood matrix quality control material is human and is prepared from the following raw materials: human whole blood, fixative, physiological saline.

3. The method according to claim 1, wherein the fixative is a PBS solution containing 2-6% (v/v) formaldehyde and 2-6% (v/v) glutaraldehyde.

4. The method of claim 1, wherein the fixative is a PBS solution containing 4% (v/v) formaldehyde and 4% (v/v) glutaraldehyde.

5. The method according to any one of claims 1 to 4, wherein the human whole blood is venous blood containing red blood cells, white blood cells and platelets of a normal human aged 18 to 50 years.

6. The method according to any one of claims 1 to 4, wherein the isolated whole blood cells are specifically: human whole blood is centrifuged at 1500g for 10min, and the upper layer is discarded, and the lower layer is the whole blood cells.

7. The method for preparing according to claim 5, wherein the separated whole blood cells are specifically: human whole blood is centrifuged at 1500g for 10min, and the upper layer is discarded, and the lower layer is the whole blood cells.

8. The method for preparing according to any one of claims 1 to 4, wherein the fixed whole blood cell obtaining immobilizate is specifically: the whole blood cells are added into the fixative and fixed for 24-48h at room temperature, and the volume ratio of the whole blood cells to the fixative is 1 (5-10).

9. The method for preparing according to claim 5, wherein the fixed whole blood cell obtaining immobilizate is specifically: the whole blood cells are added into the fixative and fixed for 24-48h at room temperature, and the volume ratio of the whole blood cells to the fixative is 1 (5-10).

10. The method for preparing as claimed in claim 6, wherein the fixed whole blood cell obtaining immobilizate is specifically: the whole blood cells are added into the fixative and fixed for 24-48h at room temperature, and the volume ratio of the whole blood cells to the fixative is 1 (5-10).

11. The method for preparing as claimed in claim 7, wherein the fixed whole blood cell obtaining immobilizate is specifically: the whole blood cells are added into the fixative and fixed for 24-48h at room temperature, and the volume ratio of the whole blood cells to the fixative is 1 (5-10).

12. The method according to claim 8, further comprising an operation of purifying the fixture, in particular: centrifuging after the whole blood cells are fixed, removing the upper fixing agent, adding physiological saline into the lower fixing object for washing for 3-5 times, wherein the volume ratio of the fixing object to the physiological saline is 1 (5-10), and then filtering to obtain filtrate, namely the purified fixing object.

13. The preparation method according to claim 12, wherein the quality control substances for preparing the high, medium and low blood glucose concentrations are specifically: respectively adding pre-prepared blood plasma with different blood sugar concentrations into a fixed object or a purified fixed object for mixing, wherein the volume ratio of the fixed object to the blood plasma is 1 (1-2), and respectively obtaining a hypoglycemic concentration quality control product, a medium blood sugar concentration quality control product and a hyperglycemic concentration quality control product; the blood plasma with different blood sugar concentration is obtained by centrifuging venous whole blood with normal blood sugar concentration and taking supernatant, or adding glucose after taking supernatant.

14. The method of claim 13, wherein the volume ratio of the immobilizate to the plasma is 1: 1.

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