CN113340883B - Method for tracing magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator - Google Patents
Method for tracing magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator Download PDFInfo
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Abstract
The invention discloses a method for tracing a magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator, which belongs to the technical field of in-vitro diagnostic detection and comprises the following steps: (1) determining a reference detection system; (2) calibrating mother liquor of a to-be-traced calibrator; (3) Diluting the mother liquor of the calibrator into a plurality of concentration points according to the nominal concentration of the mother liquor of the calibrator, and detecting samples at the concentration points; reading the measured concentration of each concentration point sample according to the standard curve in the step (1); and combining the detection result with the nominal concentration of the mother liquor of the calibrator to draw a calibrator curve; configuring the standard substance into a plurality of concentration points in the linear range of the calibration substance curve, detecting the standard substance at each concentration point, and reading the measured concentration of the standard substance at each concentration point according to the calibration substance curve; and respectively comparing the concentration deviation of the measured concentration and the nominal concentration of the calibrator to be traced and the standard substance. The tracing method is flexible to apply, and can effectively avoid the influence on the production quality of related kits caused by source tracing disorder.
Description
Technical Field
The invention relates to the technical field of in-vitro diagnosis and detection, in particular to a method for tracing a calibrator of an in-vitro diagnostic reagent by a magnetic particle chemiluminescence method.
Background
With the development of the medical health industry, the standardization and standardization of the in vitro diagnosis industry, the precision and stability of clinical examination are more and more emphasized. The magnitude transfer of the external diagnostic reagent product calibrator is an important tool for realizing accurate and traceable clinical detection results, the influence on the detection accuracy of the external diagnostic reagent is undoubted, and the accurate and consistent diagnostic results can be ensured only by paying attention to the research on the magnitude transfer and continuously improving the inspection technology, so that the development of clinical inspection career is promoted finally, and patients are benefited. Therefore, the manufacturing enterprise needs to pay attention to the calibrator traceability, establish calibrator and reference quality value traceability programs, and assign values to the calibrators and reference products produced in each batch.
At present, a systematic tracing method is lacked in the production process of the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator, so that the accuracy of clinical detection results is low, and therefore, a tracing method of the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator is required to be provided to ensure the stability and the detection reliability of products.
Disclosure of Invention
In view of this, the invention provides a method for tracing the source of a calibrator of an in-vitro diagnostic reagent by a magnetic particle chemiluminescence method, which solves the problem of source tracing chaos of calibrators and is convenient for the production quality control of related kits.
In order to achieve the purpose, the invention adopts the following technical scheme:
the method for tracing the source of the magnetic particle chemiluminescence in-vitro diagnostic reagent calibrator comprises the following steps:
(1) Determining a reference detection system:
selecting a reference detection system for tracing for the detection of the subsequent step;
drawing a standard curve according to a standard substance in a reference detection system;
(2) Calibrating mother liquor of a calibrator to be traced:
preparing a calibrator to be traced into a calibrator mother solution; diluting the calibrator stock solution into a plurality of diluted samples; detecting each diluted sample;
reading the concentration value of each detection according to the standard curve in the step (1), calculating the corresponding concentration value of the mother liquor of the calibrator according to the concentration value of each diluted sample, and averaging to obtain the nominal concentration of the mother liquor of the calibrator;
(3) Tracing the calibrator to be traced:
31 Diluting the mother liquor of the calibrator into a plurality of concentration points according to the nominal concentration of the mother liquor of the calibrator, and detecting samples at each concentration point;
32 Reading the measured concentration of each concentration point sample in the step 31) according to the standard curve in the step (1);
33 ) combining the detection result of the step 31) with the nominal concentration of the mother solution of the calibrator to draw a calibrator curve;
configuring a standard substance of a reference detection system into a plurality of concentration points in a linear range of a calibration substance curve, detecting the standard substance at each concentration point, and reading the measured concentration of the standard substance at each concentration point according to the calibration substance curve;
34 Comparing the concentration deviation of the measured concentration of each concentration point sample of the calibration product to be traced with the nominal concentration, and comparing the concentration deviation of the measured concentration of the standard product of each concentration point with the configured concentration; and evaluating the to-be-traced calibrator according to the concentration deviation result.
The traceability method established by the invention is suitable for the to-be-traced calibrator with reference substances and without reference substances, and the to-be-traced calibrator traced by the method can be used as an enterprise working reference product for applying the method to the traceability of the kit calibrator product.
Preferably, in the step (1),
for a to-be-traced calibrator with a reference substance, taking an international reference substance or a national reference substance as a standard substance, matching with a magnetic particle chemiluminescence detection reagent and a measuring instrument, establishing a reference detection system, and drawing a standard curve;
and for the to-be-traced calibrator without the reference substance, taking the established standard detection system as a reference detection system, and drawing a standard curve by using the standard substance of the reference detection system.
The standard detection system established above may be selected from those systems recognized as being better in the industry.
Preferably, all dilution steps in the above method are diluted with the same buffer.
Preferably, in the step (2),
the experiment is repeated several times, and each time of detection changes the operator and the detection time, so as to reduce the error.
Preferably, the plurality of diluted sample concentrations in step (2) are in the middle of the concentration range of the plurality of concentration point samples in step 31).
Preferably, the deviation calculation formula in step 34) is: (M-T)/T.times.100%;
wherein M is the mean value of the measured concentration, and T is the nominal concentration or the configured concentration.
Preferably, in step 34),
the concentration deviation of the lowest concentration point except the 0 concentration point is less than +/-15 percent, and the concentration deviation of the other concentration points is less than +/-10 percent, so that the product is qualified; otherwise, the product is not qualified. The evaluation criteria can also be adjusted based on actual production requirements and feedback of the application effect of the kit.
Preferably, the method further comprises the step of evaluating the uncertainty of the calibrator to be traced, and the method comprises the following steps:
a) Standard uncertainty S due to detection 1 ：
Performing at least 2 times of detection on each concentration point sample of the to-be-traced calibrator, setting a repeated experiment on each concentration point, reading a detected concentration value according to the standard curve in the step (1), and calculating an average value and a standard deviation S 1 ；
B) Standard uncertainty S due to inter-vial non-uniformity 2 ：
Detecting samples of each concentration point of the to-be-traced calibrator, detecting a plurality of bottles at each concentration point, repeatedly detecting each bottle, reading the detected concentration value according to the standard curve in the step (1), and calculating the average value and the standard deviation S 2 ；
C) Calculating the uncertainty of each concentration sample of the calibrator to be traced:
extended uncertainty U = k × U c ；
k represents data containing a factor, generally a normal distribution, with k =2 at a 95% confidence interval;
uncertainty of synthetic standard
The calibrator to be traced, which is traced by the method for tracing the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator, can be used as an enterprise work reference, and further, the method for tracing the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator is applied to tracing the kit calibrator.
The method for tracing the source of the calibration product of the enterprise work reference calibration kit comprises the following steps:
(a) Determining a reference detection system:
selecting a reference detection system for tracing for the detection of the subsequent step;
using an enterprise working reference product as a standard product of a reference detection system, tracing the enterprise working reference product by the method for tracing the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator, and drawing a standard curve according to the enterprise working reference product;
(b) Calibrating a concentrated solution of a kit calibrator:
preparing a kit calibrator as a concentrate; diluting the concentrated solution into a plurality of diluted samples; detecting each diluted sample;
reading the concentration value detected each time according to the standard curve in the step (a), calculating the corresponding concentration value of the concentrated solution according to the concentration value of each diluted sample, and averaging to obtain the nominal concentration of the concentrated solution;
(c) Tracing the source of the kit calibrator:
c1 Diluting the concentrated solution into a plurality of concentration points according to the nominal concentration of the concentrated solution, and detecting samples at each concentration point;
c2 Reading the measured concentration of the sample at each concentration point in step c 1) according to the standard curve in step (a);
c3 B) combining the detection result of the step c 1) with the nominal concentration of the concentrated solution to draw a calibration curve of the kit;
configuring the enterprise working reference products into a plurality of concentration points in a linear range of a kit calibrator curve, detecting the enterprise working reference products at each concentration point, and reading the measured concentration of the enterprise working reference products at each concentration point according to the kit calibrator curve;
c4 Comparing the concentration deviation of the measured concentration of each concentration point sample of the kit calibrator with the nominal concentration, and comparing the concentration deviation of the measured concentration of the enterprise working reference sample of each concentration point with the configured concentration; and evaluating the kit calibrator according to the concentration deviation result.
Preferably, in step (b), each diluted sample is provided with a repeat test, each test alternating operator and test time;
the concentration of the plurality of diluted samples in the step (b) is in the middle of the concentration range of the plurality of concentration point samples in the step c 1);
the deviation calculation formula in step c 4) is: (M-T)/Tx 100%.
Wherein M is the mean value of the measured concentration, and T is the nominal concentration or the configured concentration;
in the step c 4), the concentration deviation of the lowest concentration point except the concentration point 0 is less than +/-15 percent, and the concentration deviation of the other concentration points is less than +/-10 percent, so that the product is qualified; otherwise, the product is not qualified.
Preferably, the method further comprises the step of evaluating the uncertainty of the sample at each concentration point of the kit calibrator, and the method comprises the following steps:
i) Standard uncertainty S due to detection 3 ：
Detecting each concentration point sample of the kit calibrator at least 2 times, setting repeated experiments for each concentration point, reading the detection concentration value according to the standard curve in the step (a), and calculating the average value and the standard deviation S of each concentration point sample 3 ；
ii) Standard uncertainty S due to bottle-to-bottle heterogeneity 4 ：
Detecting samples of each concentration point of a kit calibrator, detecting a plurality of bottles at each concentration point, repeatedly detecting each bottle, reading the detected concentration value according to the standard curve in the step (a), and calculating the average value and the standard deviation S of each concentration point sample 4 ；
iii) Calculating the uncertainty of sample samples of various concentrations of a kit calibrator:
extended uncertainty U' = k × U c ’；
k represents data containing a factor, generally a normal distribution, with k =2 at a 95% confidence interval;
uncertainty of synthetic standard
S 5 And (4) the uncertainty of the synthesis standard introduced for the enterprise working reference.
In conclusion, the traceability method has wide application range and flexible application, and can effectively avoid the influence of traceability disorder on the production quality of related kits.
Drawings
FIG. 1 shows the standard curve obtained in example 1.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
Example 1
The method for tracing the industrial work reference product (WRS) of follicle stimulating hormone (FSH for short, national standard product number: 150533) comprises the following steps:
(1) Determining a reference detection system:
re-dissolving the standard substance by using 1mL of buffer solution according to the requirements of the national reference substance use instruction to obtain 200mIU/mL of standard substance mother solution;
diluting the re-dissolved 200mIU/mL standard mother liquor by using a calibrator diluent to obtain a series of standard concentration points of 0, 1.5, 3.0, 15.0, 50.0 and 150.0 mIU/mL;
the obtained FSH standard substance concentration points (shown in table 1) are detected by using a biological magnetic particle chemiluminescence method FSH detection reagent matched with a CIA1200 full-automatic chemiluminescence determinator as a reference detection system, a standard curve is made, and each standard substance concentration point is used as a quality control substance.
TABLE 1
| Standard article | Concentration ofmIU/ | RLU |
| A | ||
| 0 | 14684 | |
| B | 1.5 | 198283 |
| C | 3.0 | 428332 |
| D | 15.0 | 2565676 |
| E | 50.0 | 7500412 |
| F | 150.0 | 16680540 |
(2) Calibrating the mother liquor of the enterprise working reference product:
diluting an purchased enterprise working reference product used for preparing an enterprise working reference product with an antigen (10000 mIU/mL) into 1000mI U/mL enterprise working reference product mother solution; respectively diluting the enterprise work reference product mother liquor into 3 concentration points of 5, 10 and 50m IU/mL, and respectively marking as diluted samples 1, 2 and 3;
detecting each diluted sample by using a biological magnetic particle chemiluminescence FSH detection reagent matched with a CIA1200 full-automatic chemiluminescence determinator; each point is repeatedly detected for 3 times, and different operators and measuring time are changed each time; reading the concentration value detected each time according to the standard curve in the step (1), calculating the corresponding concentration value of the enterprise working reference product mother liquor according to the concentration value of each diluted sample, wherein the calculated concentration variation Coefficient (CV) of each detected 3 sample points is required to be less than 8%, and the calculated concentration variation Coefficient (CV) of each point is required to be less than 10% after 3 times of repeated calculation.
And averaging the concentration of the enterprise working reference product mother liquor calculated according to the 9 concentration values obtained by the 3 experiments of the 3 concentration points to obtain the nominal concentration of the enterprise working reference product mother liquor.
(3) Tracing the enterprise work reference product:
31 Diluting the enterprise working reference mother liquor into 6 concentration points of 0, 1.5, 3.0, 15.0, 50.0 and 150.0mIU/mL according to the nominal concentration of the enterprise working reference mother liquor, marking as WRS-A-F, and detecting samples at the concentration points by using se:Sup>A biological magnetic particle chemiluminescence FSH detection reagent matched with se:Sup>A CIA1200 full-automatic chemiluminescence determinator;
32 Reading the measured concentration of each concentration point sample in the step 31) according to the standard curve in the step (1);
33 Drawing a calibration curve (shown in figure 1) by combining the detection result of the step 31) with the nominal concentration of the mother liquor of the enterprise working reference product; reading the measured concentration of the standard substance at each concentration point according to the calibration substance curve;
34 Comparing the concentration deviation of the measured concentration of each concentration point sample of the enterprise working reference product with the nominal concentration, and comparing the concentration deviation of the measured concentration of the standard product of each concentration point with the configured concentration; the deviation calculation formula is as follows: (M-T)/Tx 100%; wherein M is the mean value of the measured concentration, and T is the nominal concentration or the configured concentration.
And (3) evaluating the to-be-traced calibrator according to the concentration deviation result: the concentration deviation of 1.5mIU/mL concentration point is less than +/-15%, and the concentration deviation of 3.0, 15.0, 50.0 and 150.0mIU/mL concentration point is less than +/-10%, so that the product is qualified; otherwise, the product is not qualified.
And (4) bottling and filling each concentration point of the FSH enterprise working reference product which is qualified through inspection.
(4) Evaluating the uncertainty of samples at each concentration point of the enterprise working reference product:
a) Standard error due to detectionDegree of certainty S 1 ：
Detecting samples of each concentration point of the enterprise working reference product for 2 times, repeating 10 samples of each concentration point each time, reading detection concentration values according to the standard curve in the step (1), and calculating the average value and the standard deviation S of each concentration point 1 ；
B) Standard uncertainty S due to inter-vial non-uniformity 2 ：
Detecting samples of each concentration point of the enterprise working reference product, detecting 10 bottles of samples at each concentration point, repeatedly detecting each bottle for 3 times, reading the detected concentration value according to the standard curve in the step (1), and calculating the average value and the standard deviation S of each concentration point 2 ；
C) Calculating the uncertainty of each concentration point sample of the enterprise working reference product:
extended uncertainty U = k × U c ；
k represents data containing a factor, generally normal distribution, with k =2 at a 95% confidence interval;
uncertainty of synthetic standard
The results of the sample uncertainty calculations for each concentration point of the FSH enterprise working reference are shown in table 2.
TABLE 2
| FSH project | WRS-B | WRS-C | WRS-D | WRS-E | WRS-F |
| S 1 | 0.022 | 0.043 | 0.214 | 0.627 | 1.748 |
| S 2 | 0.023 | 0.053 | 0.314 | 0.832 | 3.052 |
| U c | 0.032 | 0.068 | 0.380 | 1.042 | 3.518 |
| U | 0.064 | 0.136 | 0.760 | 2.084 | 7.035 |
Example 2
The method for tracing the source of the progesterone (P for short, no standard substance) enterprise work reference product (WRS) comprises the following steps:
(1) Determining a reference detection system:
selecting a Yapei ARCHITECT i system full-automatic immune luminescence analysis system for detection in the subsequent steps;
drawing a standard curve according to a standard product in the Yapei measurement system;
(2) Calibrating the mother liquor of the enterprise working reference product:
diluting an purchased enterprise working reference product used for preparing an enterprise working reference product with antigen (100000 ng/mL) into 100ng/m L enterprise working reference product mother liquor; respectively diluting the enterprise working reference product mother liquor into 3 concentration points of 1 ng/mL, 10 ng/mL and 20ng/mL, and respectively marking the concentration points as diluted samples 1, 2 and 3;
detecting each diluted sample by using a yapei measurement system; each point is repeatedly detected for 3 times, and different operators and measuring time are changed each time; reading the concentration value detected each time according to the standard curve in the step (1), calculating the corresponding concentration value of the enterprise working reference product mother liquor according to the concentration value of each diluted sample, and requiring that the calculated concentration variation Coefficient (CV) of 3 sample points detected each time is less than 8 percent and the repeated calculated concentration variation Coefficient (CV) of each point for 3 times is less than 10 percent.
And averaging the concentration of the enterprise working reference product mother liquor calculated according to the 9 concentration values obtained by the 3 experiments of the 3 concentration points to obtain the nominal concentration of the enterprise working reference product mother liquor.
(3) Tracing the enterprise work reference product:
31 Diluting the enterprise working reference product mother liquor into 6 concentration points of 0, 0.2, 1.0, 3.0, 10.0 and 50.0ng/mL according to the nominal concentration of the enterprise working reference product mother liquor, marking as WRS-A-F, and detecting samples of the concentration points by using an Yapei measurement system;
32 Reading the measured concentration of each concentration point sample in the step 31) according to the standard curve in the step (1);
33 ) drawing a calibration curve by combining the detection result of the step 31) with the nominal concentration of the mother liquor of the enterprise working reference product; reading the measured concentration of the standard substance at each concentration point according to the calibration substance curve;
34 Comparing the concentration deviation between the measured concentration and the nominal concentration of each concentration point sample of the enterprise working reference product, and comparing the concentration deviation between the measured concentration and the configured concentration of the standard product of each concentration point; the deviation calculation formula is: (M-T)/Tx 100%; wherein M is the mean value of the measured concentration, and T is the nominal concentration or the configured concentration.
And (3) evaluating the to-be-traced calibrator according to the concentration deviation result: the concentration deviation of 0.2ng/mL concentration point is less than +/-15 percent, and the concentration deviation of 1.0 ng/mL concentration point, 3.0 ng/mL concentration point, 10.0 ng/mL concentration point and 50.0ng/mL concentration point are less than +/-10 percent, and the product is qualified; otherwise, the product is not qualified.
And (4) separating and filling the qualified P enterprise working reference product concentration points in bottles.
(4) Evaluating the uncertainty of samples of various concentration points of the enterprise working reference product:
a) Standard uncertainty S due to detection 1 ：
Detecting samples of each concentration point of the enterprise working reference product for 2 times, repeating 10 samples of each concentration point each time, reading detection concentration values according to the standard curve in the step (1), and calculating the average value and the standard deviation S of each concentration point 1 ；
B) Standard uncertainty S due to inter-vial non-uniformity 2 ：
Detecting samples of various concentration points of the enterprise working reference product, detecting 10 bottles of each concentration point, repeatedly detecting each bottle for 3 times, taking a mean value, reading the detected concentration value according to the standard curve in the step (1), and calculating the mean value and standard deviation S of each concentration point 2 ；
C) Calculating the uncertainty of each concentration point sample of the enterprise working reference product:
extended uncertainty U = k × U c ；
k represents data containing a factor, generally a normal distribution, with k =2 at a 95% confidence interval;
uncertainty of synthetic standard
The results of the calculation of the uncertainty of the samples at the concentration points of the enterprise working reference are shown in table 3.
TABLE 3
| P item | WRS-B | WRS-C | WRS-D | WRS-E | WRS-F |
| S 1 | 0.002 | 0.027 | 0.038 | 0.192 | 0.611 |
| S 2 | 0.003 | 0.056 | 0.054 | 0.326 | 0.761 |
| U c | 0.004 | 0.062 | 0.066 | 0.378 | 0.976 |
| U | 0.007 | 0.125 | 0.132 | 0.757 | 1.952 |
Example 3
A method for calibrating a FSH kit calibrator for Follicle Stimulating Hormone (FSH) enterprise Working Reference (WRS), comprising the steps of:
(a) Determining a reference detection system:
using a biological magnetic particle chemiluminescence FSH detection reagent in combination with a CIA1200 full-automatic chemiluminescence determinator as a reference detection system, samples at various concentration points of a qualified FSH enterprise working reference substance (wrs) in example 1 were detected, and a standard curve was prepared.
(b) Calibrating a concentrated solution of a kit calibrator:
diluting a kit calibrator (10000 mIU/mL) into a concentrated solution of 1000 mIU/mL; respectively diluting the concentrated solution into 3 concentration points of 5, 10 and 50mIU/mL, and respectively marking the concentration points as diluted samples 1, 2 and 3;
detecting each diluted sample by using a biological magnetic particle chemiluminescence method FSH detection reagent matched with a CIA1200 full-automatic chemiluminescence determinator; each point is repeatedly detected for 3 times, and different operators and measuring time are changed each time; reading the concentration value detected each time according to the standard curve in the step (a), calculating the corresponding concentration value of the concentrated solution according to the concentration value of each diluted sample, wherein the calculated concentration variation Coefficient (CV) of 3 sample points detected each time is required to be less than 8%, and the calculated concentration variation Coefficient (CV) of each point is required to be less than 10% after 3 times of repeated calculation.
And averaging the concentration of the enterprise working reference product mother liquor calculated according to the 9 concentration values obtained by 3 experiments of the 3 concentration points to obtain the nominal concentration of the concentrated solution of the kit calibration product.
(c) Tracing the source of the kit calibrator:
c1 Diluting the concentrated solution into 6 concentration points of 0, 1.5, 3.0, 15.0, 50.0 and 150.0mIU/mL according to the nominal concentration of the concentrated solution, marking as calibrators A-F, and detecting samples of each concentration point by using a biological magnetic particle chemiluminescence method FSH detection reagent matched with a CIA1200 full-automatic chemiluminescence determinator;
c2 Reading the measured concentration of the sample at each concentration point of step c 1) according to the standard curve in step (a);
c3 B) combining the detection result of the step c 1) with the nominal concentration of the concentrated solution to draw a calibration curve of the kit; reading the measured concentration of each concentration point sample of the FSH enterprise working reference product according to a kit calibrator curve;
c4 Comparing the concentration deviation of the measured concentration of each concentration point sample of the kit calibrator with the nominal concentration, and comparing the concentration deviation of the measured concentration of the enterprise working reference sample of each concentration point with the configured concentration; the deviation calculation formula is: (M-T)/Tx 100%; wherein M is the mean value of the measured concentration, and T is the nominal concentration or the configured concentration.
And (3) evaluating a kit calibrator according to the concentration deviation result: the concentration deviation of 1.5mIU/mL concentration point is less than +/-15 percent, and the concentration deviation of 3.0, 15.0, 50.0 and 150.0mIU/mL concentration point is less than +/-10 percent, so the product is qualified; otherwise, the product is not qualified.
And bottling and filling each concentration point of the FSH kit calibrator which is qualified in the test.
(d) And (3) evaluating the uncertainty of samples at each concentration point of the calibrator of the kit:
i) Standard uncertainty S due to detection 3 ：
Detecting samples of each concentration point of the kit calibrator for 2 times, repeating 10 times for each concentration point, reading detection concentration values according to the standard curve in the step (a), and calculating the average value and the standard deviation S of each concentration point 3 ；
ii) Standard uncertainty S due to bottle-to-bottle heterogeneity 4 ：
Detecting samples of each concentration point of a kit calibrator, detecting 10 bottles of samples of each concentration point, repeatedly detecting each bottle for 3 times, reading detection concentration values according to the standard curve in the step (a), and calculating the average value and the standard deviation S of each concentration point 4 ；
iii) And calculating the uncertainty of sample samples of various concentrations of the calibrator of the kit:
extended uncertainty U' = k × U c ’；
k represents data containing a factor, generally a normal distribution, with k =2 at a 95% confidence interval;
uncertainty of synthetic standard
S 5 Uncertainty of a synthesis standard introduced for an enterprise working reference;
the results of the calculation of the sample uncertainty at each concentration of the FSH kit calibrator are shown in table 4.
TABLE 4
| FSH project | calibrator-B | calibrator-C | calibrator-D | calibrator-E | calibrator-F |
| S 3 | 0.013 | 0.038 | 0.096 | 0.670 | 0.962 |
| S 4 | 0.018 | 0.045 | 0.134 | 0.718 | 1.042 |
| S 5 | 0.032 | 0.068 | 0.380 | 1.042 | 3.518 |
| U c ’ | 0.039 | 0.090 | 0.414 | 1.432 | 3.793 |
| U’ | 0.078 | 0.180 | 0.828 | 2.864 | 7.586 |
Claims (9)
1. The method for tracing the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator is characterized by comprising the following steps of:
(1) Determining a reference detection system:
selecting a reference detection system for tracing for the detection of the subsequent step;
drawing a standard curve according to the standard substance in the reference detection system;
(2) Calibrating mother liquor of a to-be-traced calibrator:
preparing a calibrator to be traced into a calibrator mother solution; diluting the calibrator stock into a plurality of diluted samples; detecting each diluted sample;
reading the concentration value of each detection according to the standard curve in the step (1), calculating the corresponding concentration value of the mother liquor of the calibrator according to the concentration value of each diluted sample, and averaging to obtain the nominal concentration of the mother liquor of the calibrator;
(3) Tracing the to-be-traced calibrator:
31 Diluting the mother liquor of the calibrator into a plurality of concentration points according to the nominal concentration of the mother liquor of the calibrator, and detecting samples at each concentration point;
32 Reading the measured concentration of the sample at each concentration point in the step 31) according to the standard curve in the step (1);
33 Combining the detection result of the step 31) with the nominal concentration of the mother solution of the calibrator to draw a calibrator curve;
configuring the standard substance of the reference detection system into a plurality of concentration points in the linear range of the calibration substance curve, detecting the standard substance at each concentration point, and reading the measured concentration of the standard substance at each concentration point according to the calibration substance curve;
34 Comparing the concentration deviation of the measured concentration of the to-be-traced calibrator with the nominal concentration, and comparing the concentration deviation of the measured concentration of the standard with the configured concentration; evaluating the to-be-traced calibrator according to the concentration deviation result;
in the step 34), the step of,
the concentration deviation of the lowest concentration point except the 0 concentration point is less than +/-15 percent, and the concentration deviation of the other concentration points is less than +/-10 percent, so that the product is qualified;
otherwise, the product is not qualified.
2. The method for tracing the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator according to claim 1,
in the step (1), the step (c),
for a to-be-traced calibrator with a reference substance, taking an international reference substance or a national reference substance as a standard substance, matching with a magnetic particle chemiluminescence detection reagent and a measuring instrument, establishing a reference detection system, and drawing a standard curve;
and for the to-be-traced calibrator without the reference substance, taking the established standard detection system as a reference detection system, and drawing a standard curve by using the standard substance of the reference detection system.
3. The method for tracing the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator according to claim 1,
in the step (2),
the experiment was repeated several times, changing the operator and the time of detection each time.
4. The method for tracing the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator according to claim 1,
the concentrations of the plurality of diluted samples in the step (2) are in the middle of the concentration range of the plurality of concentration point samples in the step 31).
5. The method for tracing the magnetic particle chemiluminescence method in-vitro diagnostic reagent calibrator according to claim 1,
the deviation calculation formula in step 34) is: (M-T)/Tx 100%;
wherein M is the mean value of the measured concentration, and T is the nominal concentration or the configured concentration.
6. The method for tracing the source of the calibrator for the magnetic particle chemiluminescence in-vitro diagnostic reagent according to claim 1,
the method also comprises the step of evaluating the uncertainty of samples of each concentration point of the calibrator to be traced, and the method comprises the following steps:
a) Standard uncertainty S due to detection 1 ：
Performing at least 2 times of detection on each concentration point sample of the to-be-traced calibrator, setting a repeated experiment for each concentration point, reading a detected concentration value according to the standard curve in the step (1), and calculating the average value and the standard deviation S of each concentration point sample 1 ；
B) Criteria due to bottle-to-bottle non-uniformityUncertainty S 2 ：
Detecting samples of each concentration point of the to-be-traced calibrator, detecting a plurality of bottles at each concentration point, repeatedly detecting each bottle, reading the detected concentration value according to the standard curve in the step (1), and calculating the average value and the standard deviation S of each concentration point sample 2 ；
C) Calculating the uncertainty of each concentration sample of the to-be-traced calibrator:
extended uncertainty U = k × U c ；
k represents data containing a factor, generally normal distribution, with k =2 at a 95% confidence interval;
uncertainty of synthetic standard
。
7. The method for tracing the source of the calibration product of the enterprise work reference calibration kit is characterized by comprising the following steps of:
(a) Determining a reference detection system:
selecting a reference detection system for tracing for the detection of the subsequent step;
using an enterprise working reference product as a standard product of a reference detection system, tracing the source of the enterprise working reference product by the method of any one of claims 1 to 6, and drawing a standard curve according to the enterprise working reference product;
(b) Calibrating a concentrated solution of a kit calibrator:
preparing a kit calibrator as a concentrate; diluting the concentrated solution into a plurality of diluted samples; detecting each diluted sample;
reading the concentration value detected each time according to the standard curve in the step (a), calculating the corresponding concentration value of the concentrated solution according to the concentration value of each diluted sample, and taking an average value to obtain the nominal concentration of the concentrated solution;
(c) Tracing the source of the kit calibrator:
c1 Diluting the concentrated solution into a plurality of concentration points according to the nominal concentration of the concentrated solution, and detecting samples at each concentration point;
c2 Reading the measured concentration of the sample at each concentration point in step c 1) according to the standard curve in step (a);
c3 B) combining the detection result of the step c 1) with the nominal concentration of the concentrated solution to draw a calibration curve of the kit;
configuring the enterprise working reference products into a plurality of concentration points in the linear range of the kit calibrator curve, detecting the enterprise working reference products at each concentration point, and reading the measured concentration of the enterprise working reference products at each concentration point according to the kit calibrator curve;
c4 Comparing the concentration deviation of the measured concentration of each concentration point sample of the kit calibrator with the nominal concentration, and comparing the concentration deviation of the measured concentration of the enterprise working reference sample of each concentration point with the configured concentration; evaluating a kit calibrator according to the concentration deviation result;
in the step c 4), the concentration deviation of the lowest concentration point except the concentration point 0 is less than +/-15 percent, and the concentration deviation of the other concentration points is less than +/-10 percent, so that the product is qualified; otherwise, the product is not qualified.
8. The method for the traceability of calibration kits of business reference products according to claim 7,
in the step (b), each diluted sample is provided with a repeated test, and each detection changes the operator and the detection time;
the concentration of the plurality of diluted samples in the step (b) is in the middle of the concentration range of the plurality of concentration point samples in the step c 1);
the deviation calculation formula in step c 4) is: (M-T)/Tx 100%;
wherein M is the mean value of the measured concentration, and T is the nominal concentration or the configured concentration.
9. The method for the traceability of calibration kits of business reference products according to claim 7,
the kit calibrator uncertainty evaluation method further comprises the following steps of:
i) Standard uncertainty S due to detection 3 ：
Detecting each concentration point sample of the kit calibrator for at least 2 times, setting repeated experiments for each concentration point, reading the detection concentration value according to the standard curve in the step (a), and calculating the average value and the standard deviation S 3 ；
ii) standard uncertainty S due to inter-vial inhomogeneity 4 ：
Detecting samples of each concentration point of a kit calibrator, detecting a plurality of bottles at each concentration point, repeatedly detecting each bottle, reading the detection concentration value according to the standard curve in the step (a), and calculating the average value and the standard deviation S 4 ；
iii) And calculating the uncertainty of sample samples of various concentrations of the calibrator of the kit:
extended uncertainty U' = k × U c’ ；
k represents data containing a factor, generally normal distribution, with k =2 at a 95% confidence interval;
uncertainty of synthetic standard
，S 5 And (4) the uncertainty of the synthesis standard introduced for the enterprise working reference.
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