CN112083175A - Method for correcting biochemical index by hemolytic index measured by full-automatic dry biochemical analyzer - Google Patents
Method for correcting biochemical index by hemolytic index measured by full-automatic dry biochemical analyzer Download PDFInfo
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Abstract
The invention relates to a method for correcting biochemical indexes by hemolytic indexes measured by a full-automatic dry biochemical analyzer, which comprises the following steps: preparing hemolytic liquids with different hemolytic degrees, obtaining hemolytic indexes of the detector, determining the range of dry chemical biochemical indexes, determining the dry chemical biochemical indexes of the hemolytic liquids, obtaining a measured value and the hemolytic indexes, fitting the measured value and the hemolytic indexes with the hemolytic indexes of the detector, and judging the linear trend of the hemolytic indexes, so that the dry chemical biochemical indexes which can be used for correction are determined; taking a clinical serum sample which is not hemolyzed, and detecting a dry chemical biochemical index for correction by adopting a full-automatic dry biochemical instrument to serve as a true value of the corresponding dry chemical biochemical index of the serum sample; and fitting the measured value of the dry chemical biochemical index with the real value of the dry chemical biochemical index to obtain a correction formula corresponding to the dry chemical biochemical index, and calculating the correction value. The invention can simulate the hemolysis condition in actual work more truly, and is convenient for quantifying the hemolysis degree.
Description
Technical Field
The invention relates to the field of biochemical analysis and detection, in particular to a method for correcting biochemical indexes by hemolytic indexes measured by a full-automatic dry biochemical analyzer.
Background
At present, hemolysis of a specimen is one of the factors affecting the accuracy of a test result before common analysis, and in a special case, for example, when an infant, a patient with poor blood vessel condition, a patient with Paroxysmal Nocturnal Hemoglobinuria (PNH), a patient with severe anemia, a patient in rescue, a patient with autoimmune hemolysis, a patient with intravascular hemolysis, and the like cannot be collected again or a collected sample is still hemolyzed, only a yield test procedure can be performed to continuously detect the hemolyzed sample. The interference of hemolysis on clinical biochemical assays has been extensively studied, but the specific quantitative impact of the extent of hemolysis on dry chemistry indicators and how to correct hemolysis interference is unclear. Therefore, the influence of different hemolysis degrees on the dry chemistry test project result is systematically analyzed, a standard hemolysis degree colorimetric card is prepared, and a correction formula is calculated to correct the result.
Disclosure of Invention
The invention aims to provide a method for correcting biochemical indexes based on a hemolysis index measured by a Vitros full-automatic dry biochemical analyzer, aiming at the defects in the prior art.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
a method for correcting biochemical indexes by hemolytic indexes measured by a full-automatic dry biochemical analyzer comprises the following steps:
(1) preparing hemolytic solutions with different hemolytic degrees, obtaining hemolytic indexes of the detector, determining the range of dry chemical biochemical indexes, and measuring the dry chemical biochemical indexes of the hemolytic solutions to obtain measured values and hemolytic indexes;
(2) fitting the measured value of the dry chemical biochemical index with the hemolytic index of the detector, and judging the linear trend of the measured value, so as to determine the dry chemical biochemical index which can be used for correction;
(3) taking a clinical serum sample which is not hemolyzed, and detecting the dry chemical biochemical index which is determined in the step (2) and can be used for correction by adopting a full-automatic dry biochemical instrument to serve as a true value of the corresponding dry chemical biochemical index of the serum sample;
(4) fitting the dry chemical biochemical index measured value obtained in the step (1) with the real value of the corresponding dry chemical biochemical index obtained in the step (3) to obtain a correction formula of the corresponding dry chemical biochemical index;
(5) and (4) calculating the correction value of the corresponding dry chemical biochemical index in the non-hemolytic state according to the correction formula in the step (4).
In the step (1), the collected blood sample is subjected to ultrasonic cell disruption by using a JX-650 ultrasonic cell disruption instrument, and the cells are confirmed to be completely disrupted under a microscope to prepare the erythrocyte lysate.
Further, the dry chemical biochemical index is selected from: potassium ion K, sodium ion Na, chloride ion CL, calcium Ca, phosphorus PHOS, magnesium Mg, aspartate aminotransferase AST, lactate dehydrogenase LDH, creatine kinase CK, creatine kinase isozyme MBCK-MB, alanine aminotransferase ALT, total bilirubin TBIL, total protein TP, albumin ALB, gamma glutamyl transferase GGT, amylase AMY, glucose GLU, UREA UREA, URIC acid URIC, creatinine CREA.
And (3) centrifuging the collected venous blood by using a centrifugal machine for 15 minutes, observing the color of a blood supernatant, measuring the hemolysis index H on a dry biochemical instrument after visual inspection of non-hemolysis, and selecting a sample with the H <15 as a non-hemolysis sample for standby.
Further, a VITROS full-automatic biochemical analyzer is adopted for detecting the hemolytic index.
Compared with the prior art, the invention has the beneficial effects that:
the hemolysis specimen prepared by the method more truly simulates the hemolysis condition in practical work than other literature reported modes, and is convenient for quantifying the hemolysis degree; in actual work, the specific hemolysis degree of the specimen can be judged by direct visual inspection according to a standard hemolysis degree colorimetric card provided by people, and whether blood needs to be collected again is judged; meanwhile, the hemoglobin concentration in blood plasma or blood serum can be roughly evaluated according to a standard hemolysis degree colorimetric card; the result of the non-hemolytic state can be calculated according to the correction formula provided by the invention for clinical reference, so that the delay of diagnosis and treatment of the patient due to the blood re-drawing is avoided.
Drawings
FIG. 1: hemolysis liquid sample graph of different hemolysis degree.
FIG. 2: degree of hemolysis is plotted against the hemolysis index.
FIG. 3: the K ratio and the hemolytic index are linearly related.
FIG. 4: AST ratio and hemolytic index are plotted linearly.
FIG. 5: LDH ratio and hemolytic index are plotted linearly.
Detailed Description
The present invention will be described in further detail with reference to specific examples. The following examples are intended to illustrate the invention only and are not intended to limit the scope of the invention.
Hemolysis index measured by Vitros full-automatic dry biochemical instrument
Conventionally, hemolysis degree is evaluated by manual experience, and a more accurate hemolysis index can be provided by a VITROS full-automatic biochemical analyzer (Olson medical instruments Co., Ltd.) to evaluate the hemolysis degree of a sample.
1. The VITROS full-automatic biochemical analyzer adopts a special sample sucking gun head, the liquid level height of the residual sample amount of 30ul after sample adding is higher than the height of light passing, and the stability of reading absorbance is ensured.
2. Light emitted by the light source passes through the sample and is collected by the micro-sensing fiber and transmitted into the spectrophotometer assembly, after the light is incident on the grating, the grating separates the light into individual spectral components, and then an approximately 40 degree mirror reflects the light onto the photodiode array, each pixel of the photodiode array collects light from a 3.5nm wide spectral band, and the final usable wavelength range is approximately 350nm-850 nm.
3. When the spectrophotometer takes a reading, it will read all 256 pixels in the photodiode array. The "reading" of the sample parameters is done by a number of "scans" and the reference and dark reads must be done as soon as possible after the reading, where each pixel in the reading is the average of the a/D values of all scans for that pixel.
4. The absorbance at each pixel was calculated as follows.
5. Computing instrument pixel wavelength
The wavelength at each pixel is calculated by the formula lambdai＝C(i)2+B(i)+A
Where A, B, C are the three coefficients for the spectrophotometer calibration and i is the number of pixels from zero.
6. For optimum performance, the spectrophotometer used to predict the specimen condition must be and be traced to a standard spectrophotometer.
Correcting according to the standard wavelength to obtain standard absorbance
| Pixel | Wavelength of instrument | Absorbance (old) | Standard wavelength | Absorbance (New) |
| 75 | 437.65 | 0.8874910 | 436.53 | 0.879587389 |
| 76 | 440.62 | 0.9178670 | 439.49 | 0.906309963 |
| 77 | 443.59 | 0.9414590 | 442.45 | 0.932389626 |
| 78 | 446.57 | 0.9689650 | 445.41 | 0.958277683 |
| 79 | 449.54 | 1.0028100 | 448.37 | 0.989513735 |
| … | … | … | … | … |
| 183 | 761.65 | 0.1155000 | 761.78 | 0.1154523 |
| 184 | 764.67 | 0.1145430 | 764.74 | 0.1144737 |
| 185 | 767.70 | 0.1113410 | 767.69 | 0.1113516 |
7. Predicting the course of hemolysis
a) Calculating the derivative of the standard absorbance array
b) The derivative array is multiplied by the coefficient array. The CAL coefficients were generated using a set of patient samples whose hemolysis index values were referenced to standard laboratory methods. The derivation mode of the mathematical coefficient minimizes various types of variability.
c) The products of the multiplications are added.
d) The result is a prediction of hemolysis.
Sum [ P1-Pn ] ═ hemolytic index
8. Reportable range limits
In order to ensure that the hemolysis index predicted value is in a linear relation with respect to a reference method, a reportable range of the hemolysis index predicted value is set.
2. Biochemical index correction by using hemolysis index carried by machine
(1) A clinically non-hemolyzed serum sample is used and its baseline biochemical value is determined as the true value of the sample.
(2) Samples with different degrees of hemolysis are simulated by original erythrocyte and serum, and biochemical indexes of the corresponding simulated samples are detected.
(3) Sample test result original value (making original data table, for example as follows)
(4) Deduction/calculation process for correcting biochemical indexes
1. And converting the original data of each test index at each hemolysis degree into the difference value of the index without hemolysis.
In EXCEL, making the recorded difference of biochemical index results and hemolytic index into linear relation graph, performing single-factor anova and correlation regression analysis, and screening the single-factor anova and correlation regression analysisP<0.05 and the results between the respective degrees of hemolysis have an increasing or decreasing trend and the coefficient R is linearly related2>The term of 0.90 deduces the correction formula.
2. The derivation process is illustrated by taking K, AST, LDH as an example:
2.1.1 plotting the linear relationship between the K-difference and the hemolysis index
The relationship is shown in FIG. 3.
2.1.2 linear formula in the extraction graph: y is 0.0037x-0.0435
Y is the difference between the actual measurement result (B) corresponding to the degree of hemolysis and the measurement result (a) corresponding to the absence of hemolysis;
x ═ hemolytic index (H) determined corresponding to degree of hemolysis;
conversion:
results of measurement without hemolysis (A) corresponding to the degree of hemolysis (B) -0.0037 corresponding to the hemolysis index (H) +0.0435
Namely A ═ B-0.0037 ═ H +0.0435
2.2.1 plotting AST Difference and hemolytic index Linear relationship
The relationship is shown in FIG. 4.
2.2.2 linear formula in the extraction graph: 0.0927x-0.7512
Y is the difference between the actual measurement result (B) corresponding to the degree of hemolysis and the measurement result (a) corresponding to the absence of hemolysis;
x ═ hemolytic index (H) determined corresponding to degree of hemolysis;
conversion:
hemolysis index (H) +0.9456 measured when the measurement result (A) corresponds to the degree of hemolysis, and when the measurement result (B) -0.0927 corresponds to the degree of hemolysis
Namely, A is B-0.0927H +0.7512
2.3.1 plotting LDH Difference and hemolytic index Linear relationship
The relationship is shown in FIG. 5.
2.3.2 linear formula in the extraction graph: 2.9944x-34.095
Y is the difference between the actual measurement result (B) corresponding to the degree of hemolysis and the measurement result (a) corresponding to the absence of hemolysis;
x ═ hemolytic index (H) determined corresponding to degree of hemolysis;
conversion:
hemolysis index (H) +34.095 measured when the measurement result (A) corresponds to the degree of hemolysis, and when the measurement result (B) -2.9944 corresponds to the degree of hemolysis
Namely, A is B-2.9944H +34.095
In the same way, the correction formulas of all the terms meeting the conditions can be calculated.
(5) Conclusion
Index to be adjusted downwards (adjustment formula)
② index needing to be adjusted up (adjustment formula)
Index without need of regulation
Fourthly, items can not be adjusted
(6) The verification data independent sample verifies the effectiveness of the patent claims method. For a sample of hemolysis, the results of detection and estimation on a dry biochemical analyzer are as follows:
experiments in the embodiment of the invention can show that the method of the invention has reliable theoretical basis, possible practical operation, good experimental reproducibility and strong practicability. In actual work, the specific hemolysis degree of the specimen can be directly judged by visual inspection according to the standard hemolysis degree colorimetric card provided by the invention, and whether blood needs to be collected again is judged; meanwhile, the hemoglobin concentration in blood plasma or blood serum can be roughly evaluated according to the standard hemolysis degree colorimetric card in the invention; and the result of the non-hemolytic state can be calculated according to the correction formula provided by the invention for clinical reference, so that the delay of diagnosis and treatment of the patient due to the blood re-drawing is avoided.
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention in any way, and any person skilled in the art can make any simple modification, equivalent replacement, and improvement on the above embodiment without departing from the technical spirit of the present invention, and still fall within the protection scope of the technical solution of the present invention.
Claims (5)
1. A method for correcting biochemical indexes by hemolytic indexes measured by a full-automatic dry biochemical analyzer is characterized by comprising the following steps of: the method comprises the following steps:
(1) preparing hemolytic solutions with different hemolytic degrees, obtaining hemolytic indexes of the detector, determining the range of dry chemical biochemical indexes, and measuring the dry chemical biochemical indexes of the hemolytic solutions to obtain measured values and hemolytic indexes;
(2) fitting the measured value of the dry chemical biochemical index with the hemolytic index of the detector, and judging the linear trend of the measured value, so as to determine the dry chemical biochemical index which can be used for correction;
(3) taking a clinical serum sample which is not hemolyzed, and detecting the dry chemical biochemical index which is determined in the step (2) and can be used for correction by adopting a full-automatic dry biochemical instrument to serve as a true value of the corresponding dry chemical biochemical index of the serum sample;
(4) fitting the dry chemical biochemical index measured value obtained in the step (1) with the real value of the corresponding dry chemical biochemical index obtained in the step (3) to obtain a correction formula of the corresponding dry chemical biochemical index;
(5) and (4) calculating the correction value of the corresponding dry chemical biochemical index in the non-hemolytic state according to the correction formula in the step (4).
2. The method for correcting biochemical indexes by the hemolytic index measured by the full-automatic dry biochemical analyzer according to claim 1, wherein the method comprises the following steps: in the step (1), the collected blood sample is subjected to ultrasonic cell disruption by using a JX-650 ultrasonic cell disruption instrument, and the cells are confirmed to be completely disrupted under a microscope to prepare the erythrocyte lysate.
3. The method for correcting biochemical indexes by the hemolytic index measured by the full-automatic dry biochemical analyzer according to claim 1, wherein the method comprises the following steps: the dry chemical biochemical index is selected from: potassium ion K, sodium ion Na, chloride ion CL, calcium Ca, phosphorus PHOS, magnesium Mg, aspartate aminotransferase AST, lactate dehydrogenase LDH, creatine kinase CK, creatine kinase isozyme MBCK-MB, alanine aminotransferase ALT, total bilirubin TBIL, total protein TP, albumin ALB, gamma glutamyl transferase GGT, amylase AMY, glucose GLU, UREA UREA, URIC acid URIC, creatinine CREA.
4. The method for correcting biochemical indexes by the hemolytic index measured by the full-automatic dry biochemical analyzer according to claim 1, wherein the method comprises the following steps: and (3) centrifuging the collected venous blood by using a centrifugal machine for 15 minutes, observing the color of a blood supernatant, measuring the hemolysis index H on a dry biochemical instrument after visual inspection of non-hemolysis, and selecting a sample with the H <15 as a non-hemolysis sample for standby.
5. The method for correcting biochemical indexes by the hemolytic index measured by the full-automatic dry biochemical analyzer according to claim 1, wherein the method comprises the following steps: and (3) performing hemolytic index detection by adopting a VITROS full-automatic biochemical analyzer.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112683826A (en) * | 2020-12-24 | 2021-04-20 | 广电计量检测（重庆）有限公司 | Correction method and device of calibration color plate for percutaneous jaundice tester |
| CN113504369A (en) * | 2021-06-23 | 2021-10-15 | 中山大学肿瘤防治中心（中山大学附属肿瘤医院、中山大学肿瘤研究所） | Individual correction formula for eliminating positive interference of serum nerve specific enolase detection caused by specimen hemolysis and application thereof |
| CN113945708A (en) * | 2021-10-15 | 2022-01-18 | 南方医科大学 | Urine detection method and system |
| CN116298328A (en) * | 2022-12-06 | 2023-06-23 | 北京大学第三医院（北京大学第三临床医学院） | Kit for determining blood potassium concentration of hemolysis specimen |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112683826A (en) * | 2020-12-24 | 2021-04-20 | 广电计量检测（重庆）有限公司 | Correction method and device of calibration color plate for percutaneous jaundice tester |
| CN113504369A (en) * | 2021-06-23 | 2021-10-15 | 中山大学肿瘤防治中心（中山大学附属肿瘤医院、中山大学肿瘤研究所） | Individual correction formula for eliminating positive interference of serum nerve specific enolase detection caused by specimen hemolysis and application thereof |
| CN113945708A (en) * | 2021-10-15 | 2022-01-18 | 南方医科大学 | Urine detection method and system |
| CN116298328A (en) * | 2022-12-06 | 2023-06-23 | 北京大学第三医院（北京大学第三临床医学院） | Kit for determining blood potassium concentration of hemolysis specimen |
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