A calibration method for a four-item coagulation detection system and a four-item coagulation detection method

Abstract

The present invention relates to the field of bioanalysis detection technology, and in particular to a calibration method for four coagulation tests. The calibration method first uses gradient concentration samples to establish an initial calibration function of a system to be calibrated and a reference system; then, through the detection of clinical samples, the deviation of the detection results between the reference system and the system to be calibrated is obtained, and correction coefficients K and B are obtained. The initial calibration function is corrected using the calibration coefficients to obtain the final calibration function of the system to be calibrated. Experiments show that using a calibrated system to detect samples to be tested has the advantages of high accuracy, simple operation, saving time and reagents, etc.

Description

Technical Field

[0001] The present invention relates to the technical field of biological analysis and detection, and in particular to a calibration method for a blood coagulation four-item detection system and a blood coagulation four-item detection method. Background Art

[0002] The four coagulation tests include thrombin time (TT), prothrombin time (PT), and activated partial thromboplastin time (ACT). APTT , fibrinogen FIB. At present, most coagulation tests on the market need to be paired with special coagulation analyzers, and large coagulation analyzers are not suitable for areas with poor health and medical conditions due to their high prices and high subsequent maintenance costs. Therefore, they have great limitations and cannot be widely used. Small coagulation testing systems have relatively low purchase costs and use costs, but in order to ensure the accuracy and stability of the test results, manufacturers need to conduct a time-consuming and labor-intensive calibration process before leaving the factory. Moreover, after replacing the four coagulation test reagents from different batches or manufacturers, the test results may have large deviations. Therefore, it is of great significance to establish a test method with good universality, small detection deviation and high accuracy for the four coagulation tests. Summary of the invention

[0003] In view of this, the present invention provides a calibration method for a coagulation four-item detection system and a coagulation four-item detection method. The system calibrated by the method has small detection deviation and high accuracy.

[0004] The present invention provides the following technical solutions:

[0005] A calibration method for a coagulation four-item detection system, using a reference system to calibrate a system to be calibrated, comprising the following steps:

[0006] (1) Establishing the correlation function between the system to be calibrated and the reference system as the initial calibration function of the system to be calibrated;

[0007] (2) testing a plurality of clinical samples on a reference system to obtain a plurality of reference values; testing the plurality of clinical samples on a system to be calibrated to obtain a plurality of signal values, and substituting the plurality of signal values ​​into an initial calibration function to calculate a plurality of test values;

[0008] (3) calculating the deviation between the reference value and the test value of each clinical sample in step (2) to obtain multiple deviations of multiple clinical samples;

[0009] (4) Using the average value of the multiple deviations as a correction coefficient, the initial calibration function described in step (1) is corrected to obtain a final calibration function of the system to be calibrated.

[0010] In the calibration method of the present invention, the step (1) specifically comprises:

[0011] Four samples of coagulation with gradient concentrations are taken and tested on the system to be calibrated and the reference system respectively to obtain reference values ​​and test signal values. Function fitting is performed on the reference values ​​and test signal values ​​using the same samples as the corresponding relationship to obtain an initial calibration function.

[0012] In some specific embodiments, the initial calibration function is: y=kx+b.

[0013] In the calibration method of the present invention, in step (3), the deviation includes a first deviation and / or a second deviation; the deviation is the first deviation, or is the second deviation, or includes the first deviation and the second deviation.

[0014] The first deviation is the ratio of the reference value to the test value, where the ratio = reference value / test value; the second deviation is the difference between the reference value and the test value, where the difference = reference value - test value.

[0015] The present invention uses the average value of the first deviation and / or the second deviation as the correction coefficient K and the correction coefficient B respectively, and uses the correction coefficient K and / or the correction coefficient B to correct the initial calibration function to obtain the final calibration function of the system to be calibrated. Specifically, the following five schemes are included:

[0016] Solution 1: Use the calibration coefficient K to correct the initial calibration function of step (1) to obtain calibration function 1 as the final calibration function of the system to be calibrated. The calibration method specifically includes the following steps:

[0017] Steps (1) to (2) are as described above;

[0018] Step (3): calculating the first deviation between the reference value and the test value of each clinical sample in step (2), and obtaining a plurality of first deviation values ​​of a plurality of clinical samples;

[0019] Step (4): using the average value of the plurality of first deviation values ​​as the correction coefficient K, correcting the initial calibration function to obtain the calibration function 1 of the system to be calibrated.

[0020] In some specific embodiments, step (4) specifically includes:

[0021] The average value of multiple first deviation values ​​is taken as the correction coefficient K, and the y value of the initial calibration function (y=kx+b) is corrected using the correction coefficient K, that is, the correction coefficient K is substituted into the initial calibration function y=kx+b to obtain the calibration function 1 of the system to be calibrated: y=K(kx+b)=Kkx+Kb.

[0022] Solution 2 (the deviation is the second deviation): Use the calibration coefficient B to correct the initial calibration function of step (1) to obtain calibration function 2 as the final calibration function of the system to be calibrated. The calibration method specifically includes the following steps:

[0023] Steps (1) to (2) are as described above;

[0024] Step (3): calculating the second deviation between the reference value and the test value of each clinical sample in step (2), and obtaining a plurality of second deviation values ​​of a plurality of clinical samples;

[0025] Step (4): using the average value of the plurality of second deviation values ​​as a correction coefficient B, correcting the initial calibration function to obtain a calibration function 2 of the system to be calibrated.

[0026] In some specific embodiments, step (4) specifically includes:

[0027] The average value of multiple second deviation values ​​is used as the correction coefficient B, and the y value of the initial calibration function (y=kx+b) is corrected using the correction coefficient B. That is, the correction coefficient B is substituted into the initial calibration function y=kx+b to obtain the calibration function 2 of the system to be calibrated: y=kx+b+B.

[0028] Solution 3 (the deviation includes the first deviation and the second deviation): firstly use the calibration coefficient K to correct the initial calibration function of step (1) to obtain calibration function 1, and then use the calibration coefficient B to correct the calibration function 1 to obtain calibration function 3 as the final calibration function of the system to be calibrated. The calibration method specifically includes the following steps:

[0029] Steps (1) to (2) are as described above;

[0030] Step (3): calculating the first deviation between the reference value and the test value of each clinical sample in step (2) to obtain a plurality of first deviation values ​​of the plurality of clinical samples; calculating the second deviation between the reference value and the test value of each clinical sample in step (2) to obtain a plurality of second deviation values ​​of the plurality of clinical samples;

[0031] Step (4): Using the average value of the multiple first deviation values ​​as the correction coefficient K, and using the average value of the multiple second deviation values ​​as the correction coefficient B, first use the correction coefficient K to calibrate the initial calibration function, and then use the correction coefficient B to calibrate the calibration result again to obtain the calibration function 3 of the system to be calibrated.

[0032] In some specific embodiments, step (4) specifically includes:

[0033] An average value of the plurality of first deviation values ​​is used as a correction coefficient K, and an average value of the plurality of second deviation values ​​is used as a correction coefficient B;

[0034] First, the y value of the initial calibration function (y=kx+b) is corrected using the correction coefficient K to obtain calibration function 1; then the y value of the calibration function 1 is corrected using the calibration coefficient B. This process means: first, the correction coefficient K is substituted into the initial calibration function y=kx+b to obtain calibration function 1: y=K(kx+b)=Kkx+Kb; then, the calibration coefficient B is substituted into calibration function 1 to obtain calibration function 3: y=K(kx+b)+B=Kkx+Kb+B.

[0035] Solution 4 (the deviation includes the first deviation and the second deviation): first use the calibration coefficient B to correct the initial calibration function of step (1) to obtain calibration function 1, and then use the calibration coefficient K to correct the calibration function 1 to obtain calibration function 4 as the final calibration function of the system to be calibrated. The calibration method specifically includes the following steps:

[0036] Steps (1) to (2) are as described above;

[0037] Step (3): calculating the first deviation between the reference value and the test value of each clinical sample in step (2) to obtain a plurality of first deviation values ​​of the plurality of clinical samples; calculating the second deviation between the reference value and the test value of each clinical sample in step (2) to obtain a plurality of second deviation values ​​of the plurality of clinical samples;

[0038] Step (4): using the average value of the plurality of first deviation values ​​as the correction coefficient K, and using the average value of the plurality of second deviation values ​​as the correction coefficient B; firstly using the correction coefficient B to correct the initial calibration function, and then using the correction coefficient K to correct the correction result again, to obtain the calibration function 4 of the system to be calibrated.

[0039] In some specific embodiments, step (4) specifically includes:

[0040] An average value of the plurality of first deviation values ​​is used as a correction coefficient K, and an average value of the plurality of second deviation values ​​is used as a correction coefficient B;

[0041] First, the y value of the initial calibration function (y=kx+b) is corrected using the correction coefficient B to obtain calibration function 2; then the y value of the calibration function 2 is corrected using the calibration coefficient K. This process means: first, the correction coefficient B is substituted into the initial calibration function y=kx+b to obtain calibration function 2: y=kx+b+B; then, the calibration coefficient K is substituted into calibration function 2 to obtain calibration function 4: y=K(kx+b+B)=Kkx+Kb+KB.

[0042] Solution 5 (the deviation includes the first deviation and the second deviation): The initial calibration function of step (1) is corrected using calibration coefficients K and B respectively to obtain calibration function 1 and calibration function 2, and the calibration function with the smaller deviation compared with the reference system is selected as the final calibration function. The calibration method specifically includes the following steps:

[0043] Steps (1) to (2) are as described above;

[0044] Step (3): Calculate the second deviation between the reference value and the test value of each clinical sample in step (2) to obtain multiple second deviation values ​​of multiple clinical samples;

[0045] Step (4): Use calibration coefficients K and B to calibrate the initial calibration function of step (1) to obtain calibration function 1 and calibration function 2; calculate the deviations of calibration function 1 and calibration coefficient 2 relative to the reference system, and use the calibration function with the smaller deviation (calibration function 2 or calibration function 3) as the final calibration function.

[0046] Furthermore, the step (4) specifically includes steps S1 to S3:

[0047] S1, using the average value of the plurality of first deviations as a correction coefficient K, correcting the initial calibration function to obtain a calibration function 1 of the system to be calibrated;

[0048] S2, using an average value of the plurality of second deviations as a correction coefficient B, to correct the initial calibration function, and obtain a calibration function 2 of the system to be calibrated;

[0049] S3, calculating the third deviation of calibration function 1 relative to the reference system and the fourth deviation of calibration function 2 relative to the reference system; taking the calibration function corresponding to the lower value of the third deviation and the fourth deviation as the final calibration function.

[0050] In step S3, the third deviation and the fourth deviation are calculated by the following formulas:

[0051] The third deviation = (test value of calibration function 1 - reference value of reference system) × 100% / reference value of reference system;

[0052] Fourth deviation=(test value of calibration function 2−reference value of reference system)×100% / reference value of reference system.

[0053] Specifically, step S3 includes the following steps:

[0054] S3-1. Clinical samples with multiple gradient concentrations are tested on the reference system and the system to be calibrated to obtain multiple reference values ​​and multiple signal values;

[0055] S3-2, respectively substituting the plurality of signal values ​​into the calibration function 1 obtained in step S1 and the calibration function 2 obtained in step S2, and correspondingly obtaining a plurality of first test values ​​and a plurality of second test values;

[0056] S3-3, calculating the third deviation between the first test value of each clinical sample and its reference value, and taking the average value thereof; similarly, calculating the fourth deviation between the second test value of each clinical sample and its reference value, and taking the average value thereof;

[0057] S3-4. Compare the two average values ​​obtained in step S3-3, and use the calibration function (calibration function 1 or calibration function 2) corresponding to the smaller average value as the final calibration function of the system to be calibrated.

[0058] After calibrating the system to be calibrated using the above calibration method, the final calibration functions of PT, APTT, TT, and FIB of the system to be calibrated are obtained respectively.

[0059] The present invention also provides a method for detecting four items of coagulation, including:

[0060] Testing the four coagulation samples on a calibrated system to obtain test results;

[0061] The calibrated system is calibrated using any one of the aforementioned calibration methods.

[0062] The calibration method provided by the present invention first uses gradient concentration samples to establish the initial calibration function of the system to be calibrated and the reference system; then, through the detection of clinical samples, the deviation of the detection results between the reference system and the system to be calibrated is obtained, and the correction coefficients K and B are obtained, and then the initial calibration function is specifically corrected using the calibration coefficients, and finally the final calibration function of the system to be calibrated is obtained. Experiments show that the detection of the sample to be tested using the calibrated system of the present invention has the advantages of small deviation, high accuracy, simple operation, saving time and reagents, etc. DETAILED DESCRIPTION

[0063] The present invention provides a calibration method for a coagulation four-item detection system and a coagulation four-item detection method. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It is particularly important to point out that all similar replacements and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0064] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0065] The present invention will be further described below in conjunction with embodiments:

[0066] Example 1

[0067] The present invention provides a calibration method for a coagulation four-item test system, which has the advantages of high accuracy, simple operation, saving time and reagents, etc., and specifically comprises the following steps:

[0068] a) Take 50 clinical plasma samples with different PT / APTT / TT / FIB values ​​(i.e. gradient concentration samples) and test them with the first reagent on the reference system and the second reagent on the system to be calibrated. The concentration distribution of each index of the gradient samples is shown in Table 1, and the test data is shown in Table 2.

[0069] The test results of the two are subjected to linear regression processing to obtain the linear correlation equation 1 (y=kx+b), where y is the test result of the reference coagulation test system (referred to as the reference system), x is the test result of the system to be calibrated, and k and b are linear correlation coefficients.

[0070] The linear correlation equations 1 of the four indicators are obtained respectively, that is, four calibration equations:

[0071] APTT: y1=0.8945x + 10.801, R2=0.9832;

[0072] PT: y1=1.5002x – 3.9981, R2=0.9873;

[0073] TT: y1=0.9743x + 0.4075, R2=0.989;

[0074] FIB: y1=-0.9475x + 3.3604, R2=0.985.

[0075] Table 1

[0076]

[0077] Table 2

[0078]

[0079]

[0080] b) 10 clinical plasma samples with different PT / APTT / TT / FIB values ​​were taken and placed on the reference system corresponding to the first reagent and on the system to be calibrated corresponding to the second reagent for testing, and the reference value Y and signal value X1 were obtained respectively. The test results of the four indicators are shown in Table 3.

[0081] Substitute the signal value X1 as X into equation 1 in step a) to obtain the test value Y1;

[0082] The reference value Y and the test value Y1 of each clinical sample were compared, and the first deviation Y / Y1 and the second deviation Y-Y1 were calculated respectively. The results are shown in Table 3;

[0083] Calculate the average values ​​of the first deviations of the four indicators respectively to obtain their respective correction coefficients K (see Table 4);

[0084] The average values ​​of the second deviations of the four indicators, PT, APTT, TT and FIB, were calculated respectively to obtain their respective correction coefficients B (see Table 4);

[0085] Substitute the correction coefficient K into the initial calibration function (y=kx+b) to obtain the calibration functions 1 of the four indicators: y=K*(kx+b)=Kkx+Kb;

[0086] Substitute the correction coefficient B into the initial calibration function (y=kx+b) to obtain the four indexes’ more accurate functions 2: y=kx+b+B;

[0087] The specific equation is as follows:

[0088] APTT: calibration function y1=0.8853x+10.6898, calibration function y2=0.8945x+10.521;

[0089] PT: calibration function y1=1.4589x – 3.8880, calibration function y2=1.5002x – 4.308;

[0090] TT: calibration function y1=0.9563x+0.4000, calibration function y2=0.9743x+0.068;

[0091] FIB: calibration function y1 = -0.9254x + 3.2820, calibration function y2 = -0.9475x + 3.303.

[0092] Table 3

[0093]

[0094] Table 4

[0095]

[0096] c) Use 10 clinical samples to verify standard function 1 and standard function 2. By comparing the deviation with the reference system, the one with the smallest deviation is taken as the final calibration function of coagulation. The specific process is as follows:

[0097] 10 clinical samples to be tested were tested on the reference system and the system to be calibrated, respectively, and the reference value Y# and the test value X1# were obtained in turn. The test value X1# was substituted into the calibration function 1 and the calibration function 2 as the X value, respectively, to obtain the test values ​​Y1# and Y2# (see Table 5);

[0098] The test values ​​Y1# and Y2# of all the clinical samples to be tested are obtained according to the above method, and the test values ​​Y1# and Y2# of each clinical sample are compared with the reference value Y#, and the third deviation of the calibration function 1 and the reference system, and the fourth deviation of the calibration function 2 and the reference system are calculated according to the following formulas:

[0099] The third deviation = (Y1#-Y#) / Y#×100%, the fourth deviation = (Y2#-Y#) / Y#×100%;

[0100] The average values ​​of the third deviation and the fourth deviation of all clinical samples to be tested were calculated, and the results are shown in Table 6. The calibration function with the smallest average value was taken as the final calibration function of the coagulation indexes PT, APTT, TT, and FIB.

[0101] Table 5

[0102]

[0103] Table 6 Deviation results from the reference system

[0104]

[0105] According to the deviation results in Table 6, the calibration function with the smaller deviation is selected as the final calibration function of the coagulation index, and the final calibration functions of the four indicators are obtained respectively:

[0106] APTT: y1=0.8853x + 10.6898;

[0107] PT: y2=1.5002x – 4.308;

[0108] TT: y1=0.9563x + 0.4000;

[0109] FIB:y2=-0.9475x + 3.303.

[0110] Example 2

[0111] The final calibration function determined in Example 1 was used to calibrate the results of 10 clinical samples (different from the samples in Example 1). The four coagulation reagents of the standard test system and the specified test system were the same as the corresponding reagents in Example 1 and were from the same batch of reagents. The results are shown in Table 7.

[0112] Table 7

[0113]

[0114] It can be seen from Table 7 that the average CV values ​​of the test results of the system calibrated by the calibration method of the present invention and the coagulation test system are both less than 5%, indicating that the calibration method of the present invention is suitable for the calibration of the four coagulation test system, and the detection of the four coagulation indicators by the calibrated system after the calibration of the present invention has small deviation and high accuracy.

[0115] The above embodiments verify and illustrate the correction process and technical effects of Scheme 5. For Schemes 1, 2, 3 and 4, based on the correction calculation process of each scheme, it can be theoretically deduced that the correction method is to correct the test value closer to the reference value. Therefore, it can be inferred that the correction schemes of Schemes 1 to 4 of the present invention all have the effect of improving the accuracy of system calibration.

[0116] The above are only preferred embodiments of the present invention. It should be pointed out that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A calibration method for a coagulation four-item detection system, using a reference system to calibrate the system to be calibrated, characterized in that: The following steps are involved: (1) Establishing a correlation function between the system to be calibrated and the reference system as the initial calibration function of the system to be calibrated; (2) Testing multiple clinical samples on a reference system to obtain multiple reference values; Testing the multiple clinical samples on the system to be calibrated to obtain multiple signal values, and substituting the multiple signal values ​​into the initial calibration function to calculate multiple test values; (3) respectively calculating the first deviation and the second deviation between the reference value and the test value of each clinical sample in step (2) to obtain a plurality of first deviations and second deviations of the plurality of clinical samples; The first deviation is the ratio of the reference value to the test value, and the second deviation is the difference between the reference value and the test value; (4) comprising steps S1 to S3: S1, using the average value of the plurality of first deviations as a correction coefficient K, correcting the initial calibration function to obtain a calibration function 1 of the system to be calibrated; S2, using an average value of the plurality of second deviations as a correction coefficient B, to correct the initial calibration function, and obtain a calibration function 2 of the system to be calibrated; S3, calculating a third deviation of calibration function 1 relative to the reference system and a fourth deviation of calibration function 2 relative to the reference system; The calibration function corresponding to the lower value of the third deviation and the fourth deviation is taken as the final calibration function.

2. The calibration method according to claim 1, characterized in that: The step (1) is: Four samples of coagulation with gradient concentrations are taken and tested on the system to be calibrated and the reference system respectively to obtain reference values ​​and test signal values. Function fitting is performed on the reference values ​​and test signal values ​​using the same samples as the corresponding relationship to obtain an initial calibration function.

3. The calibration method according to claim 1, characterized in that: The third deviation and the fourth deviation are calculated by the following formulas: Third deviation=(test value of calibration function 1 - reference value of reference system)×100% / reference value of reference system; Fourth deviation=(test value of calibration function 2−reference value of reference system)×100% / reference value of reference system.

4. The calibration method according to claim 1, characterized in that: The step S3 specifically includes: S3-1. Clinical samples with multiple gradient concentrations are tested on the reference system and the system to be calibrated to obtain multiple reference values ​​and multiple signal values; S3-2, respectively substituting the plurality of signal values ​​into the calibration function 1 obtained in step S1 and the calibration function 2 obtained in step S2, and correspondingly obtaining a plurality of first test values ​​and a plurality of second test values; S3-3, calculating the third deviation between the first test value of each clinical sample and its reference value, and taking the average value thereof; similarly, calculating the fourth deviation between the second test value of each clinical sample and its reference value, and taking the average value thereof; S3-4. Compare the two average values ​​obtained in step S3-3, and use the calibration function corresponding to the smaller average value as the final calibration function of the system to be calibrated.

5. A four-item coagulation detection method, characterized in that: include: Testing the four coagulation samples on a calibrated system to obtain test results; Wherein, the calibrated system is calibrated using the calibration method described in any one of claims 1 to 4.

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