CN113740523A - TSH multi-system assignment method based on magnitude traceability - Google Patents

TSH multi-system assignment method based on magnitude traceability Download PDF

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CN113740523A
CN113740523A CN202111073064.0A CN202111073064A CN113740523A CN 113740523 A CN113740523 A CN 113740523A CN 202111073064 A CN202111073064 A CN 202111073064A CN 113740523 A CN113740523 A CN 113740523A
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张顺利
王清涛
魏星
成斐
高瑞丰
王�华
莫玉
贾婷婷
王默
尹弘毅
张瑞
岳育红
崔瑞芳
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Beijing Center For Clinical Laboratory
Beijing Chaoyang Hospital
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Beijing Huanuo Aomei Gene Biotechnology Co ltd
Beijing Chaoyang Hospital
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Abstract

The invention discloses a TSH multi-system assignment method based on magnitude traceability. The assignment method assigns the candidate TSH standard substance by adopting a TSH international standard substance quantity transmission mode, and the system comprises an instrument for quantitatively detecting TSH and used reagents, such as a TSH detection system in a clinical laboratory. The TSH candidate standard substance of human serum source, which is researched and obtained by the assignment method of the invention, has rich raw material sources. The candidate standard substance is subjected to combined assignment through a plurality of detection systems, and the international standard substance WHO NIBSC 81/565 is used for magnitude transmission, so that the candidate standard substance is expected to be used for calibration or calibration verification of a conventional detection method in a clinical laboratory and assignment of a conventional calibrator of a reagent manufacturer, and can play a role in the indoor quality evaluation and accuracy verification plan of the conventional detection system, and the accuracy and consistency of TSH detection results of different laboratories and different detection systems are improved and monitored.

Description

TSH multi-system assignment method based on magnitude traceability
Technical Field
The invention relates to a TSH multi-system assignment method based on magnitude traceability, and belongs to the field of standard substance assignment.
Background
Thyroid Stimulating Hormone (TSH) is a glycoprotein Hormone secreted by adenohypophysis basophils and consists of 211 amino acids, and its molecular structure consists of noncovalent α, β subunit heterodimers, wherein the Hormone is specific for biological and immunological functions and is β subunit, and α subunit is substantially identical to α subunit of Luteinizing Hormone (LH), Follicle Stimulating Hormone (FSH) and Human Chorionic Gonadotropin (HCG). TSH can promote secretion of T3 (triiodothyronine) and T4 (thyroxine), and is the most sensitive index for judging thyroid gland function under negative feedback of T3 and T4, and the level of the index should be closely monitored in the course of thyroid gland disease treatment.
Immunoassay of TSH has been changed by Radioimmunoassay (RIA), immunoradiometric Assay (IRMA), Enzyme Linked Immunosorbent Assay (ELISA), Chemiluminescence Immunoassay (CLIA), Electrochemiluminescence Immunoassay (Electrochemiluminescence Immunoassay, ECLIA), Time-resolved Fluoroimmunoassay (TrFIA), and Chemiluminescence Immunoassay (CLEIA). TSH manufacturers have large detection results, and Beckmann Access 2 is about 20 times higher than Siemens Immunite 2000 in some TSH detection values with low concentration. Recent studies have shown that TSH varies less between methods at normal concentrations and more at abnormal concentrations. The comparison research of 6 TSH detection methods by domestic Malong et al discovers that the methods have different differences, and discovers that the correlation between the radioimmunoassay and the radioimmunoassay with an automatic detection system is poor (r is 0.38-0.41).
The purpose of standardization is to make the test results accurate and consistent for each laboratory/manufacturer. The main content of standardization work is to trace the source to the international system of units (SI) through a high-level primary Reference substance and/or a Reference Measurement Program (RMP), the core is magnitude tracing, namely, a reliable Reference system is established, and the accuracy is transferred to clinical routine analysis through the system, so that the routine result is traced to the accuracy provided by the Reference system. TSH belongs to protein test items, the detection principle of the items is mostly an immune method, RMP is not acknowledged, and only a consistent scheme can be adopted if results are comparable. The TSH detection item has international universal standard substances, so that the TSH detection item is convenient for manufacturers and researchers to use. But only the reference substances of the interoperability can fully exert their effects. Recent international studies on ERM 470 have also demonstrated a lack of interoperability among ceruloplasmin, and this is considered one of the causes of inconsistency in the detection results between systems, with similar guesses for TSH studies. Domestic Tongqing et al adopt intercommunicating human serum enzymology reference substance to calibrate the detection instrument of second-level and third-level hospitals in Beijing, and find that the difference of detection results among laboratories is obviously reduced after calibration.
Since the world health organization uk national biologicals certification institute TSH international standard (WHONIBSC 81/565) is a high-concentration TSH lyophilized powder, each manufacturer must perform a series of multiple dilutions in tracing the source with the standard. As early as the year 2008, it was,
Figure BDA0003261105410000021
a study scheme of assignment of a dilution method to a protein test project is proposed by Blirup-Jensen et al. They used CRM 470 to accurately transfer its values to the manufacturer's primary calibrators and working calibrators or quality controls and demonstrated the examples. For direct target value transfer between serum matrices, the scheme is to assign values to the reference substances by performing a calibration curve fit based on multiple measurements of 6-point dilutions of the reference substances and the target substances. The transfer protocol required repeated measurements daily for several days. Both rewet and dilution are controlled by weighing to reduce uncertainty in assignment (target shift). In an open system, a reference substance may be used as a calibrator and a target substance as a sample. The ratio of the concentrations of the two species (with or without matrix effect) was directly estimated by a single regression plot. If there is no matrix effect, the regression line will pass through zero with a slope equal to the ratio of the two species concentrations. In closed systems, dedicated commercial calibrators, reference substances and target substances must be used as samples for analysis.
At present, TSH standard substances without human serum matrix are not used for the accuracy verification of indoor quality evaluation mechanisms due to the reasons that international standard substances are expensive, lack of interchangeability, long in transportation period, insufficient in supply of goods sources and the like, and the TSH standard substances cannot meet the requirements of increasing national enterprises on TSH high-order standard substances. Therefore, the research and the scientific multi-system assignment of the TSH standard substance of the human serum matrix are extremely important.
Disclosure of Invention
The invention aims to provide a TSH multi-system assignment method based on magnitude traceability, which assigns a candidate TSH standard substance by adopting a TSH international standard substance to carry out magnitude transmission.
The "system" in the present invention includes an instrument for quantitatively detecting TSH and a reagent used, such as a clinical laboratory TSH detection system.
The TSH multi-system assignment method based on magnitude traceability provided by the invention comprises the step of assigning a candidate TSH standard substance by adopting a TSH international standard substance magnitude transmission mode.
Specifically, the assignment method includes the following steps:
(a1) collecting and subpackaging in-vitro individual human serum with different TSH concentrations according to 3 detection amounts (milliliters and ml) of samples required by n mainstream domestic TSH detection systems to form a serum tray, wherein n is 5-10;
the in vitro individual human serum is a single human serum with clear and transparent appearance except for jaundice, hemolysis, lipemia, chyle and other abnormal characters;
(a2) diluting the TSH international standard substance into 2-3 concentration series with different alternative diluents (also called matrixes), randomly inserting the series into the serum tray in the step (a1), and carrying out interchangeability research;
the TSH international standard substance is WHO NIBSC 81/565;
(a3) carrying out interchangeability evaluation on the TSH international standard substances in different alternative diluents, screening to obtain alternative diluents with interchangeability among the TSH detection systems as matrixes, and then carrying out the following steps:
(a4) diluting the TSH international standard substance by using the substrate obtained by screening to obtain a series of standard series solutions of the TSH international standard substance with different concentrations, and calculating to obtain the authentication concentration of the TSH international standard substance in each standard series solution of the TSH international standard substance;
(a5) separately determining the concentration of TSH international standard substance in each of the diluted solutions obtained in step (a4) using the TSH detection system;
(a6) and (b) performing linear fitting on the concentrations of the TSH international standard substances obtained in the step (a4) and the step (a5) to obtain a TSH assignment standard curve, namely, achieving assignment of the TSH candidate standard substance with interchangeability.
In the above assignment method, in step (a1), the TSH detection system includes siemens ADVIA CentaurXP, siemens Immulite 2000, architecett i2000sr, beckman DXI800, soliton XL, riedman CI 1000, robas 601, and Autolumo a2000 plus;
the initial concentration range of TSH detected by adopting Siemens ADVIA CentauerXP of the serum in vitro is 0.09 mu IU/mL-84.03 mu IU/mL.
In the assignment method described above, in step (a2), the alternative diluent comprises TSH 0. mu.IU/mL pooled Human Serum (HSP) serum0) The serum albumin-free solution can be prepared by the following steps of (1) preparing a BSA-containing Hanks 'balanced salt solution (abbreviated as Hanks solution), a BSA-containing Du's phosphate buffer solution (abbreviated as DPBS) or a BSA-containing normal saline.
"high, low and medium" respectively means higher than the upper limit of the reference interval, the lower limit of the reference interval and the reference interval, the concentration level is about 10 times, for example, the high value is 40 mu IU/mL, the median value is 4 mu IU/mL, the low value is 0.4 mu IU/mL;
the "interchangeability", also known as interoperability, is an important property of a reference substance, meaning that there is an equivalent numerical relationship between the reference substance and a series of representative sample measurements between different measurement procedures.
In the assignment method, in the step (a3), the judgment criteria for the interchangeability evaluation are that the reference substance (including the TSH international standard substance and the TSH candidate standard substance in the different standard series solutions) is within two 95% confidence line ranges above and below the Deming regression line drawn by the TSH detection values of the individual sera obtained by the two TSH detection systems;
the diluent obtained by screening is TSH 0 mu IU/mL mixed human serum and Hanks' balanced salt solution containing BSA (Hanks solution for short).
In the assignment method, in the step (a4), the following steps are adopted to obtain the certified concentration of the TSH international standard substance in the diluted solution:
(A) measuring the density of the TSH international standard substance stock solution or the standard series solution with higher concentration and the standard series solution of the TSH international standard substance, and respectively marking as rho1And ρ2；
(B) Obtaining the authentication concentration of TSH of the standard series solution of the TSH international standard substance according to the formula (1), and marking as C;
Figure BDA0003261105410000041
wherein, C1Expressing the concentration of TSH in the TSH international standard substance stock solution or the higher concentration standard series solution; m1Representing the quality of the TSH international standard substance or higher concentration standard series solution; rho1Representing the density of the TSH international standard substance stock solution or higher concentration standard series solution; m2Representing the quality of alternative diluent in the standard series solution of the TSH international standard substance; rho2Represents the density of the standard series solution of the TSH international standard substance.
Wherein, in the steps (A) and (B), the densities of the TSH international standard substance standard series solution and the TSH international standard substance stock solution or the higher concentration standard series solution are measured according to the following steps:
(b1) the total mass of the weighing container and the gun head of the sample-adding gun is recorded as m1；
(b2) Absorbing a sample to be detected with the volume v by adopting the gun head of the sample adding gun, then placing the sample in the container, weighing the whole sample and recording the whole sample as m2；
(b3) Calculating the density of the sample to be detected according to a formula (2);
ρto be measured＝(m2-m1)/v (2)。
The TSH candidate standard substance of human serum source, which is researched and obtained by the assignment method of the invention, has rich raw material sources. The candidate standard substance is subjected to combined assignment through a plurality of detection systems, and the international standard substance WHO NIBSC 81/565 is used for magnitude transmission, so that the candidate standard substance is expected to be used for calibration or calibration verification of a conventional detection method in a clinical laboratory and assignment of a conventional calibrator of a reagent manufacturer, and can play a role in the indoor quality evaluation and accuracy verification plan of the conventional detection system, and the accuracy and consistency of TSH detection results of different laboratories and different detection systems are improved and monitored.
Drawings
FIG. 1 shows interchangeability between multiple systems for TSH international standard substances diluted in multiple matrices (4 matrices in Table 2 on the left and 2 matrices used for assignment therein on the right), where the criteria for interchangeability are within the two 95% confidence lines (dotted lines) above and below the Deming regression line drawn for individual serum TSH measurements.
FIG. 2 is a assignment curve of TSH candidate standard substances (L1-L11) at different concentrations using Siemens ADVIA CentauerXP and two kinds of substrate diluted TSH international standard substances (left side substrate is mixed human serum with TSH concentration of 0 μ IU/mL, and right side substrate is Hanks' balanced salt solution containing bovine serum albumin (abbreviated as Hanks solution)).
FIG. 3 is a plot of assignment curves of TSH candidate standards (L1-L11) at different concentrations using Lidmann CI 1000 and two matrix dilutions of TSH international standard (left matrix is mixed human serum with a TSH concentration of 0 μ IU/mL, right matrix is Hanks' balanced salt solution with bovine serum albumin (Hanks solution for short)).
FIG. 4 is a assignment curve of TSH candidate standard substances (L1-L11) at different concentrations using Roche Cobas 601 and two kinds of substrate diluted TSH international standard substances (left substrate is mixed human serum with TSH concentration of 0 μ IU/mL, right substrate is Hanks' balanced salt solution (Hanks solution for short)) containing bovine serum albumin.
FIG. 5 shows assignment curves of TSH international standard substance diluted with Mike IS 1200 and two kinds of matrices as TSH candidate standard substance (L1-L11) of different concentrations (left matrix IS mixed human serum with TSH concentration of 0 μ IU/mL, right matrix IS Hanks' balanced salt solution (Hanks solution for short) containing bovine serum albumin).
FIG. 6 is a value curve of TSH candidate standard substances (L1-L11) with different concentrations, which are assigned to TSH international standard substances diluted by Maxumi 2000plus and two kinds of matrixes (left matrix is mixed human serum with TSH concentration of 0 μ IU/mL, and right matrix is Hanks' balanced salt solution (Hanks solution for short) containing bovine serum albumin).
FIG. 7 shows the interchangeability of two substrate-diluted TSH international standards between Mike IS 1200 and the New Productivity Maglumi2000plus system (using mixed human serum instead of individual serum dishes).
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The TSH multi-system assignment method based on magnitude traceability, which is provided by the invention, assigns candidate TSH standard substances by adopting a TSH international standard substance magnitude transmission mode, and specifically comprises the following steps:
(a1) the remaining sera (ethical approval 2018-2-26-1) after completion of the tests of 29 patients with different TSH concentrations were collected from the kyoto yang hospital affiliated with the university of capital medical science. Serum from each sample was divided into 8 portions according to the manufacturer's requirements and tested 3 times in 8 different systems (see table 1). The samples were free of hemolysis, lipemia and jaundice and the initial concentration of TSH measured using the Siemens ADVIA CentauerXP immunoassay ranged from 0.09. mu.IU/mL to 84.03. mu.IU/mL.
TABLE 1 TSH detection System basic information
Figure BDA0003261105410000061
AMR, analyzing the measuring range; RR, reference range; ECL, electrochemiluminescence; CL, chemiluminescence; CMIA, chemiluminescent microparticle immunoassay.
(a2) The compatibility study was conducted by selecting 4 kinds of the TSH international standard substance, diluting the TSH international standard substance into 2-3 concentration series with high or low TSH concentration, and randomly inserting the TSH international standard substance into the serum dish (a 1).
The interchangeability, also called interoperability, is an important attribute of a reference substance (here, TSH international standard substance diluted by 4 kinds of diluents to different concentrations, hereinafter referred to as international standard substance standard series solution, and also referred to as TSH candidate standard substance), and means that there is an equivalent numerical relationship between the reference substance and a series of representative samples among different measurement procedures. The judgment standard with interchangeability is that the reference substance is in two 95% credible line ranges above and below a Deming regression line drawn by an individual serum TSH detection value, and meanwhile, the judgment standard can be measured by adopting the bias related to interchangeability, and the smaller the absolute value of the bias is, the better the interchangeability is. The formula of 4 kinds of diluent is shown in
TABLE 2, interchangeability associated bias, is shown in Table 3, which is calculated as follows.
Figure BDA0003261105410000062
Wherein, CMean value, otherIs the TSH concentration mean value, C obtained by detecting the international standard substance standard series solution by other detection systems (evaluation methods)Mean value, cenThe mean value of the TSH concentration obtained by detecting international standard substance standard series solutions by Siemens ADVIA CentaurXP (comparative method), wherein beta and alpha respectively refer to the slope and intercept of Deming regression.
TABLE 2 basic information on dilutions of International Standard substances
Figure BDA0003261105410000063
HSP0The TSH concentration is 0 mu IU/mL of mixed human serum; BSA, bovine serum albumin (Sigma, B2064) was present at a concentration of 46 g/L.
(a3) Will be at different rarefactionsCarrying out interchangeability evaluation on international standard substances in the release solution; selecting two (HSP)0And Hanks' balanced salt solution with BSA (Hanks solution for short)) were compatible between most systems the following assignment experiments (fig. 1 and table 3) were performed using the assignment curves for ADVIA CentaurXP, see fig. 2, with smaller differences in the slope of the assignment curves in the left and right columns.
TABLE 3 interchangeability related bias in different dilutions (%), with ADVIA CentaurXP as control system international standard
Figure BDA0003261105410000071
HSP0The TSH concentration is 0 mu IU/mL mixed human serum; IS, TSH international standard substance; BSA, bovine serum albumin; hanks' balanced salt solution (Hanks solution for short); DPBS, dubi phosphate buffer (DPBS for short); N.S, saline.
(a4) Adopting HSP as TSH international standard substance0Diluting with Hanks' balanced salt solution (Hanks solution for short) containing BSA to obtain standard series solutions of TSH international standard substances, and calculating to obtain the TSH authentication concentration (tables 4 and 6) in each diluted solution, wherein the density is shown in tables 5 and 7.
Obtaining the TSH authentication concentration in serial diluted solutions of TSH international standard substances with different concentrations, and calculating the TSH authentication concentration in the standard serial solutions of the TSH international standard substances according to the following formula:
Figure BDA0003261105410000081
wherein, C1Expressing the concentration of TSH in the TSH international standard substance stock solution or the higher concentration standard series solution; m1Representing the quality of the TSH international standard substance or higher concentration standard series solution; rho1Representing the density of the TSH international standard substance stock solution or higher concentration standard series solution; m2Representing the quality of alternative diluent in the standard series solution of the TSH international standard substance; rho2The densities of the standard series solutions of the TSH international standard substance are indicated.
Diluting TSH international standard substance with mixed Human Serum (HSP)0) Or Hanks' balanced salt solution (Hanks liquid for short) containing BSA, wherein the TSH concentration of the serum and the Hanks liquid is 0 mu IU/mL; HSP0The appearance of the product is clear and transparent, except for abnormal characters such as jaundice, hemolysis, lipemia and chyle.
The TSH concentration in the TSH international standard substance stock solution is obtained by weighing TSH international standard substance (WHO NIBSC 81/565) diluted by Phosphate Buffered Saline (PBS) solution containing Bovine Serum Albumin (BSA), and the concentration is 1181.356 μ IU/mL.
TABLE 4 TSH International Standard substance Standard series solution concentration preparation (the dilution used was HSP)0)
Figure BDA0003261105410000082
The standard series 13 added is the standard series of the higher concentration, i.e. the expected concentration of 68 μ IU/mL.
TABLE 5 TSH International Standard substance Standard series solution Density measurement (HSP as diluent used)0)
Figure BDA0003261105410000091
TABLE 6 TSH International Standard substance Standard series solution concentration preparation (Hank solution as the diluent)
Figure BDA0003261105410000092
The standard series 26 added is the standard series with the higher concentration, i.e. the sample with the expected concentration of 68 μ IU/mL.
TABLE 7 TSH International Standard substance Standard series solution Density measurements (Hanks' solution as diluent used)
Figure BDA0003261105410000101
(a5) The concentration of TSH in each of the diluted solutions obtained in step (a4) was measured using 8 TSH quantitative determination systems, respectively.
(a6) And (3) aiming at each TSH quantitative detection system in the 8 TSH quantitative detection systems, performing linear fitting by adopting the authentication concentration of the TSH in each diluted solution obtained in the step (a4) and the determination concentration of the TSH in each diluted solution obtained in the step (a5) to obtain a TSH assignment standard curve, and thus assigning values to the 11 TSH candidate standard substances.
To illustrate the significance of international standard substance interchangeability on TSH candidate standard substance assignment consistency. Dilution series (HSP) in Table 4 were used0) The 7 detection systems used were ADVIA CentaurXP, Immulite 2000, Architect i2000sr, DXI800, Liaison XL, CI 1000 and Cobas 601, and 5 detection systems in dilution series (Hanks liquid) in Table 6 were used including ADVIA CentaurXP, DXI800, Cobas 601, Autolumo A2000 and CI 1000. From Table 8, it can be seen that when CI 1000 was diluted with poor interchangeability Hanks 'solution (mean bias 28.29%, see also V-2 in FIG. 1) or when Cobas 601 was used with poor interchangeability HSP' s0(average bias-19.51%, see also VI-2 in FIG. 1) the difference between the two was large at dilution. At the same time, a poorly interchangeable substrate was used (i.e., Hanks solution for CI 1000 and HSP for Cobas 601)0) Substrates with good compatibility when assigned (i.e., CI 1000 uses HSP's)0Copas 601 using Hanks liquid) gave greater differences (see tables 8 and 9), and in addition, the slope of the curves for the left and right columns gave greater differences in the slope of the curves as seen in fig. 3 and 4. Finally, 10 detection methods of 8 systems are adopted for assignment, and the average value is taken as a target value or a provisional value (see table 9). It should be noted that 11 TSH candidate standard substances (L1 to L11) having different concentrations are compatible with each other (left column in FIG. 1), which is an important attribute of the candidate standard substances. In addition, because the concentration levels of the TSH candidate standard substances are greatly different, L1-L2, L3-L4, L5-L7, L8-L10 and L11 respectively adopt different assignment curves (see fig. 2-4), and the left side and the right side of each figure are respectively provided with different assignment curves (see fig. 2-4)The top 5 figures correspond to L1-L2, L3-L4, L5-L7, L8-L10 and L11, respectively.
TABLE 8 assignment bias of the same detection System with different substrates
Figure BDA0003261105410000111
L1-L2, L3-L4, L5-L7, L8-L10 and L11 respectively adopt different assignment curves (see FIGS. 3-4), and 5 graphs from top to bottom on the left side and the right side of each graph respectively correspond to L1-L2, L3-L4, L5-L7, L8-L10 and L11.
TABLE 98 detection systems 10 methods assignment of TSH candidate Standard substances (target values)
Figure BDA0003261105410000121
(a7) To verify HSP0And Hanks liquid IS applicable to TSH international standard substance assignment and the range of multi-system assignment IS enlarged, two detection systems (IS 1200, Magumi 2000plus, as shown in Table 1) of Mike and new industries are selected for diluting the TSH international standard substance and assigning 11 TSH candidate standard substances with interchangeability and different concentration levels. The TSH concentrations in the diluted solutions were calculated from the standard series of TSH international standard substances (tables 10 and 12), and the calculation method was the same as (a4), and the densities are shown in tables 11 and 13. The assignment methods are the same as (a5) - (a 6).
TABLE 10 TSH International Standard substance Standard series solution concentration formulations (HSP)0)
Figure BDA0003261105410000131
The number 13 standard series of the higher concentration standard series solution was added, i.e. the sample with the expected concentration of 60 μ IU/mL.
TABLE 11 TSH International Standard substance Standard series solution Density assay (HSP)0)
Figure BDA0003261105410000132
TABLE 12 TSH International Standard substance Standard series solution concentration formulations (Hanks liquid)
Figure BDA0003261105410000141
The higher concentration standard series 26 standard series was added, i.e. the expected concentration of 60 μ IU/mL.
TABLE 13 TSH International Standard substance Standard series solution Density measurements (Hanks' solution)
Figure BDA0003261105410000142
(a8) Two systems (Maglumi 2000plus and IS 1200) on two substrates (HSP's)0And Hanks 'solution) and target value bias in table 8 are shown in table 14 although IS 1200 assigned a mean bias of-12.4% (-14.5% to-10.2%) and slightly more than 11.73% for international standards diluted with Hanks' solution, none of the assigned results exceeded 3 standard deviations (3SD), and the assigned curve slopes were all slightly different (left and right columns of fig. 5). Therefore, the method is incorporated into the final assignment system. The values for Maglumi2000plus were all significantly different for the values assigned using the Hanks liquid diluted international standard (left and right columns in FIG. 6), the bias for the values assigned was large, with a mean bias of-38.3% (-44.7% to-31.8%), greater than the minimum allowable bias derived based on biological variability of 11.73%, and the values assigned were outside of 3SD, mainly due to the poor interchangeability of the Hanks liquid diluted international standard between the two systems (see FIG. 7) and the large matrix effect (Table 15).
Figure BDA0003261105410000151
Table 15 IS compared with IS 1200Systemic international standard substance in HSP0Bias associated with interchangeability in Hanks' solution (%)
Figure BDA0003261105410000161
HSP0The TSH concentration is 0 mu IU/mL mixed human serum; hanks liquid, Hanks' balanced salt solution.
(a9) Finally, 13 assignment methods using 10 detection systems are adopted, the assignment results of the TSH candidate standard substances with 11 levels are shown in Table 16, 3-fold standard deviation (3SD) is used as an outlier judgment standard, and the outliers are not removed.
(a10) Uncertainty of fit (u) during assignmentver) The calculation formula is as follows:
Figure BDA0003261105410000162
Figure BDA0003261105410000163
wherein S isRThe standard deviation of the assignment curve; a. thejDetecting concentration values for the TSH; b is0An assigned curve intercept is obtained; b is1The slope of the assignment curve; rho is the authenticated concentration; p is the determination frequency of the TSH candidate standard substance, and n is the total determination frequency of the TSH international standard substance.
(a11) The candidate standard substance expansion uncertainty calculation formula is as follows:
Figure BDA0003261105410000164
(where k is 2, which is an expansion factor; u)verFitting process of standard curve and uncertainty of international standard substance; u. ofimpStandard deviation assigned for multiple systems)
(a12) The final assignment results were 0.191-44.985 μ IU/mL with an expanded uncertainty range of 0.039-3.523 μ IU/mL, as shown in Table 17.
Figure BDA0003261105410000171
TABLE 17 Final assignment of TSH candidate Standard substances
Figure BDA0003261105410000181
The invention adopts mixed human serum with TSH concentration of 0 mu IU/mL and Hanks liquid to dilute international certified standard substance WHO NIBSC 81/565 to make an assignment curve, adopts 13 methods of 10 detection systems to assign values to TSH candidate standard substances, has accurate and consistent assignment results, and provides a new idea for TSH standardization and consistency work.
The TSH candidate standard substance of human serum source developed by the invention has rich raw material sources. The candidate standard substance is subjected to joint assignment through a plurality of detection systems, and the TSH international standard substance WHO NIBSC 81/565 is adopted for magnitude transmission, so that the candidate standard substance has good interchangeability, is expected to be used for calibration or calibration verification of a conventional detection method in a clinical laboratory and assignment of a conventional calibrator of a reagent manufacturer, can play a role in the indoor quality evaluation and accuracy verification plan of the conventional detection system, and improves and monitors the accuracy and consistency of TSH detection results of different laboratories and different detection systems.
The invention can provide necessary data support for promoting the clinical inspection quality of TSH in Beijing area and even nationwide, improving the effectiveness of the work of preventing and treating diseases, saving and effectively utilizing sanitary resources, and can possibly generate certain economic benefit and social benefit in the future.

Claims (10)

1. A TSH multi-system assignment method based on magnitude traceability comprises the step of assigning a candidate TSH standard substance in a manner of magnitude transmission by adopting a TSH international standard substance.
2. The assignment method of claim 1, wherein: the assignment method comprises the following steps:
(a1) adopting different TSH detection systems to respectively detect the in vitro individual human serum with different TSH concentrations to form a serum tray;
(a2) diluting the TSH international standard substance into a series of 2-3 concentrations with different alternative diluents, randomly inserting the TSH international standard substance into the serum tray in the step (a1), and carrying out interchangeability study;
(a3) carrying out interchangeability evaluation on the TSH international standard substances in different alternative diluents, screening to obtain alternative diluents with interchangeability among the TSH detection systems as matrixes, and then carrying out the following steps:
(a4) diluting the TSH international standard substance by using the substrate obtained by screening to obtain a series of standard series solutions of the TSH international standard substance with different concentrations, and calculating to obtain the authentication concentration of the TSH international standard substance in each standard series solution of the TSH international standard substance;
(a5) separately determining the concentration of TSH international standard substance in each of the diluted solutions obtained in step (a4) using the TSH detection system;
(a6) and (b) performing linear fitting on the certified concentrations of the TSH international standard substances obtained in the step (a4) and the step (a5) to obtain a TSH assignment standard curve, namely, assignment of the TSH candidate standard substances with different concentrations and interchangeability is realized.
3. The assignment method of claim 2, wherein: in step (a1), the TSH detection system comprises ADVIA CentaurXP, Immulite 2000, DXI800, Autolumo a2000plus, CI 1000, Cobas 601, Architect i2000sr, and Liaison XL;
the TSH concentration of the isolated individual human serum in the step (a1) is 0.09 mu IU/mL-84.03 mu IU/mL.
4. A method of assigning values according to claim 2 or 3, characterized in that: in step (a2), the alternative diluent comprises a mixed human serum with a TSH concentration of 0 μ IU/mL, a Hanks 'balanced salt solution containing bovine serum albumin, a Du's phosphate buffer solution containing BSA, or a physiological saline solution containing BSA.
5. The assignment method of any one of claims 2-4, wherein: in the step (a3), the judgment standard of the interchangeability evaluation is that the reference substance is in the upper and lower 95% credible line ranges of a Deming regression line drawn by the individual serum TSH detection values obtained by the two TSH detection systems;
the reference substances comprise TSH international standard substances and TSH candidate standard substances in different standard series solutions;
the alternative dilutions obtained by screening were pooled human serum with a TSH concentration of 0. mu.IU/mL and a Hanks' balanced salt solution containing BSA.
6. The assignment method of any one of claims 2-5, wherein: in the step (a4), the certified concentration of the TSH international standard substance in the standard series solution of the TSH international standard substance is obtained by the following steps:
(A) measuring the densities of the TSH international standard substance stock solution or the higher concentration standard series solution and the TSH international standard substance standard series solution, and respectively marking as rho1And ρ2；
(B) Obtaining the authentication concentration of TSH of the standard series solution of the TSH international standard substance according to the formula (1), and marking as C;
Figure FDA0003261105400000021
wherein, C1Expressing the concentration of TSH in the TSH international standard substance stock solution or the higher concentration standard series solution; m1Representing the quality of the TSH international standard substance or higher concentration standard series solution; rho1Representing the density of the TSH international standard substance stock solution or higher concentration standard series solution; m2Representing the quality of alternative diluent in the standard series solution of the TSH international standard substance; rho2Represents the density of the standard series solution of the TSH international standard substance.
7. The assignment method of claim 5, wherein: in the steps (A) and (B), the densities of the TSH international standard substance standard series solution and the TSH international standard substance stock solution or the higher concentration standard series solution are measured according to the following steps:
(b1) the total mass of the weighing container and the gun head of the sample-adding gun is recorded as m1；
(b2) Absorbing a sample to be detected with the volume v by adopting the gun head of the sample adding gun, then placing the sample in the container, weighing the whole sample and recording the whole sample as m2；
(b3) Calculating the density of the sample to be detected according to a formula (2);
ρto be measured＝(m2-m1)/v (2)。
8. The assignment method of any one of claims 1-7, wherein: the TSH international standard substance is WHO NIBSC 81/565.
9. The assignment method of any one of claims 1-8, wherein: the TSH candidate standard substance with interchangeability is mixed human serum with clear and transparent appearance and abnormal properties such as jaundice, hemolysis, lipemia, chyle and the like.
10. A TSH candidate standard substance obtained by the method of assigning a value according to any one of claims 1 to 9.
CN202111073064.0A 2021-09-14 2021-09-14 TSH multi-system assignment method based on magnitude traceability Pending CN113740523A (en)

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