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CN-121995066-A - Immunosuppressant accurate monitoring treatment kit containing steroid hormone

CN121995066ACN 121995066 ACN121995066 ACN 121995066ACN-121995066-A

Abstract

The invention relates to the technical field of in-vitro diagnostic reagents, in particular to an immunosuppressant accurate monitoring treatment kit containing steroid hormone, which comprises a combined detection and correction method for accurate monitoring of the immunosuppressant containing steroid hormone, and comprises the following steps of 1, collecting clinical samples to be detected and sub-packaging each collected sample into two equal parts; and step 2, performing immunological quantitative determination on the first equivalent sample by adopting an immunological kit to obtain an immunological determination value, and step 3, performing reference quantitative determination on the second equivalent sample corresponding to the step 2 by adopting a liquid chromatography-tandem mass spectrometry to obtain a mass spectrometry determination value, wherein the mass spectrometry is performed quantitatively by using an isotope labeled internal standard corresponding to each target analyte according to a standardized sample pretreatment flow. According to the invention, the high flux and the operation convenience of an immunization method are ensured, and meanwhile, the accuracy equivalent to that of a mass spectrometry method is provided, so that a reliable basis is provided for clinical personalized medication.

Inventors

  • CAO JIANBIAO
  • LI ANJIE

Assignees

  • 深圳华腾生物医疗电子有限公司

Dates

Publication Date
20260508
Application Date
20251216

Claims (8)

  1. 1. The immunosuppressant accurate monitoring treatment kit containing the steroid hormone is characterized by comprising a combined detection and correction method for accurate monitoring of the immunosuppressant containing the steroid hormone, and comprises the following steps of: step 1, collecting clinical samples to be tested and sub-packaging each collected sample into two equal parts; step 2, performing immunological quantitative determination on the first sample by adopting an immunological kit to obtain an immunological determination value; Step 3, performing reference quantitative determination on the second equivalent sample corresponding to the step 2 by adopting a liquid chromatography-tandem mass spectrometry to obtain a mass spectrometry measurement value, wherein the mass spectrometry is quantitatively performed by using an isotope labeled internal standard corresponding to each target analyte according to a standardized sample pretreatment flow; step 4, calculating the intercept and the slope of a linear conversion model by adopting a doming regression method based on the paired data obtained in the step 2 and the step 3 so as to meet the requirement that any immune method measured value can be converted into a corresponding mass spectrum equivalent value through a linear conversion rule that the mass spectrum equivalent value is equal to the intercept plus the slope multiplied by the immune method measured value; And 5, in daily detection, carrying out mass spectrum verification in the step 3 on samples meeting the triggering conditions according to a preset triggering rule, merging newly-added paired data into a model update queue, reconstructing and version-storing the intercept and the slope according to a preset period or the accumulated sample number, so as to convert an immunometric measurement value into a mass spectrum equivalent value and notice the version and the source of the used model in a detection report.
  2. 2. The method for combined detection and calibration of immunosuppressant precise monitoring of steroid hormone according to claim 1, wherein the internal standard for mass spectrometry is an isotope labeled internal form corresponding to each target analyte one by one, and is used for calibrating sample pretreatment recovery rate, sample introduction difference and matrix effect.
  3. 3. A combined detection and calibration method for accurate detection of immunosuppressants containing steroid hormones as claimed in claim 1, wherein the sample pretreatment comprises protein precipitation with an organic solvent to remove protein interference, followed by purification and enrichment of the supernatant by solid phase extraction for improved recovery and selectivity of mass spectrometry.
  4. 4. The method for combined detection and correction of immunosuppressant accurate monitoring of steroid hormone according to claim 1, wherein the intercept and slope meet the following acceptance criteria when establishing, wherein the difference sequence is calculated based on paired data, a Bland Altman method is adopted to obtain a consistency interval, and the paired sample ratio in the consistency interval is not lower than ninety five percent; simultaneously calculating the average relative bias and making the average relative bias within the allowable total error range, wherein the allowable total error is preferably plus or minus fifteen percent; in addition, the number of pairing samples used in the initial establishment of the conversion model is not less than sixty.
  5. 5. The steroid hormone-containing immunosuppressant precision monitoring treatment kit of claim 1, wherein the kit comprises at least an antibody reagent set for immunological assay, a multi-point calibrator matched with clinical sample matrix, a low/medium/high three-grade quality control, an interference detection reagent for identifying or blocking a xenotropic antibody, and an information interface or electronic file for receiving, storing or accompanying intercept and slope of the mass spectrometry-to-immunology conversion model; the instructions or electronic files attached to the kit indicate that when a reagent lot is changed or a preset sample size or a preset period is reached, mass spectrometry correction should be performed and the updated intercept and slope written back or issued according to the method of claim 1.
  6. 6. The steroid hormone containing immunosuppressant precision monitoring process kit of claim 1, comprising a software product for automatically converting an immunoassay to mass spectrometry equivalent and managing the conversion model, the software containing executable instructions to perform the functions of receiving and managing a paired dataset, performing a doming regression based on the received data to calculate the intercept and slope of the linear conversion model, calculating the Bland-Altman index, average bias and consistency interval and determining if the model passes according to preset allowable overall error and coverage criteria, versioning the passed model and providing the function of writing version information back to the kit electronic interface or laboratory information system, automatically determining if mass spectrometry verification is required for a single sample or sample lot according to preset trigger rules in daily operation.
  7. 7. The steroid hormone containing immunosuppressant precision monitoring treatment kit of claim 1, wherein the preset trigger rules comprise at least one of: a) triggering mass spectrometry verification if the immunometric measurement value approaches or falls within a preset critical interval of a clinical critical decision threshold defined by a clinical guideline or laboratory, b) triggering mass spectrometry verification if the relative change of the current immunometric measurement value and its previous measurement value of the same patient exceeds a preset threshold of the laboratory, c) triggering mass spectrometry verification if the current or current lot quality control index exceeds the preset threshold, or the detection of a foreign antibody or other interference is positive, d) performing random spot check according to a preset sampling proportion to trigger mass spectrometry verification.
  8. 8. The steroid hormone containing immunosuppressant precision monitoring processing kit of claim 1, wherein the model updating and drift monitoring comprises automatically reconstructing a doming regression model and generating a new version when the number of accumulated incorporated mass spectrometry verification pairing samples reaches a laboratory preset threshold or reaches a preset time period, performing online monitoring on continuously obtained average bias or model parameters by adopting a statistical process control algorithm, automatically issuing a reconstruction or manual examination alarm if a statistically significant drift is detected by the monitoring algorithm, and writing the new intercept and slope obtained after reconstruction back in the new version and replacing the previous version for conversion of subsequent immunological measurement values into mass spectrometry equivalent values.

Description

Immunosuppressant accurate monitoring treatment kit containing steroid hormone Technical Field The invention relates to an immunosuppressant accurate monitoring and processing kit containing steroid hormone, in particular to an immunosuppressant accurate monitoring and processing kit containing steroid hormone, belonging to the technical field of in-vitro diagnostic reagents. Background Immunosuppressants containing steroid hormones are required to be monitored for long-term and accurate drug concentration in clinical organ transplantation, autoimmune diseases and chronic inflammatory disease treatment to guide personalized dosing and avoid toxic side effects; At present, immunological methods (such as chemiluminescence, radioimmunoassay, enzyme-exemption and the like) are mainly used for quantitatively detecting the hormone immunosuppressant in clinic, the method has the advantages of high automation degree, large detection flux, simple operation and the like, but the detection result is easy to be interfered by antibody cross reaction, anisotropic antibody, metabolite and matrix components, so that the problems of incomplete identification, quantitative bias and the like frequently occur among steroid hormones with similar structures in a low concentration interval, and the method is characterized in that the method is used as a recognized gold standard in multi-target simultaneous identification, low concentration quantification, metabolite differentiation and specificity, but the method has high equipment cost, complex operation and high maintenance requirement and is not suitable for the conventional high flux detection scenes of all laboratories; In order to make up the result difference between an immunological method and LC-MS/MS, partial laboratories adopt two methods for parallel measurement and manually establish a quantitative comparison relation, but the existing methods generally have the problems of irregular data processing, lack of unified standard of a conversion model, dependence of a manual experience on a updating period, incapacity of automatically identifying the failure of the conversion model, difficulty in processing the difference between batches and pretreatment difference, and the like, the existing kit does not provide a conversion model which can be directly used for correction of an immunological method-mass spectrometry, and also has no unified regression mode, quality control acceptance standard or consistency evaluation flow, and the steps of protein precipitation, solid phase extraction and the like in the pretreatment process of a sample cause uncertainty of recovery rate and matrix effect, further increase the deviation between the immunological method and the mass spectrometry, and in addition, the imperfect recognition mechanism of the anisotropic antibody interference also affects the accuracy of the result of the immunological method, so that the accurate monitoring and treatment kit for the immunosuppressant containing steroid hormone is needed to be improved to solve the problems. Disclosure of Invention The invention aims to provide an immunosuppressant accurate monitoring processing kit containing steroid hormone, which aims to solve the problem that an integral system capable of standardizing, verifying and traceably converting an immunological measurement result and an LC-MS/MS measurement result is lacking, wherein the integral system comprises a unified sample pretreatment flow, an interference suppression mechanism, a regression model construction method, bias and consistency evaluation standards and an automatic model management mechanism, so that an immunological result cannot reliably replace a mass spectrum result at a clinical decision point. In order to achieve the above purpose, the present invention provides the following technical solutions: The immunosuppressant accurate monitoring treatment kit containing steroid hormone comprises a combined detection and correction method for accurate monitoring of the immunosuppressant containing steroid hormone, and comprises the following steps: step 1, collecting clinical samples to be tested and sub-packaging each collected sample into two equal parts; step 2, performing immunological quantitative determination on the first sample by adopting an immunological kit to obtain an immunological determination value; Step 3, performing reference quantitative determination on the second equivalent sample corresponding to the step 2 by adopting a liquid chromatography-tandem mass spectrometry to obtain a mass spectrometry measurement value, wherein the mass spectrometry is quantitatively performed by using an isotope labeled internal standard corresponding to each target analyte according to a standardized sample pretreatment flow; step 4, calculating the intercept and the slope of a linear conversion model by adopting a doming regression method based on the paired data obtained in the step 2 and the step 3 so as to