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Internal standard correction-based immune precise quantitative analysis method and special kit thereof

A kit and reagent technology, applied in the direction of analyzing materials, analyzing materials by electromagnetic means, measuring devices, etc., can solve problems such as stability and consistency affecting the accuracy of quantitative results

Inactive Publication Date: 2020-07-28
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The stability and consistency of this reaction will directly affect the accuracy of quantitative results.

Method used

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  • Internal standard correction-based immune precise quantitative analysis method and special kit thereof
  • Internal standard correction-based immune precise quantitative analysis method and special kit thereof
  • Internal standard correction-based immune precise quantitative analysis method and special kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0093] Example 1. Accurate immune kit based on internal standard correction and its use method

[0094] 1. Accurate immunoassay kit based on internal standard calibration

[0095] Accurate immunoassay kit based on internal standard calibration includes detection probe, internal standard probe, 96-well plate or magnetic microspheres, antigen standard, PBS buffer, washing solution (PBST solution), acid dissociation solution (nitric acid aqueous solution) .

[0096] 1. Detection probe

[0097] Use natural isotopes with relatively high isotopic abundance 153 Eu-labeled β2 microglobulin to obtain detection probes. Specific steps are as follows:

[0098] (1) DTPA (S-2-(4-Isothiocyanatobenzyl)-diethylenetriamine pentaacetic acid) compound and 153 Eu rare earth ions were dissolved in sodium acetate buffer (0.5M, pH=5.8) at a molar ratio of 1:1.2, and reacted at room temperature for 30 minutes to obtain a macrocycle-rare earth chelate.

[0099] (2) Dissolve the β2 microglobulin a...

Embodiment 2

[0117] Example 2. The precise quantitative analysis method of immunity based on internal standard correction and the precision detection of traditional methods

[0118] 1. Immunoaccurate quantitative analysis method based on internal standard correction

[0119] The β2-MG antigen standard substance of 1.0 μg / mL was used as the sample to be tested, and the content of β2-MG in the sample to be tested was detected using the kit and its detection method in Example 1, and the detection was repeated 6 times each time. 4 trials. according to obtained 151 Eu and 153 The signal strength of Eu and figure 2 The standard curve for the internal standard calibration method is shown, and the results for all replicate experiments calculated by the internal standard calibration method are obtained.

[0120] 2. Traditional method

[0121] The β2-MG antigen standard of 1.0 μg / mL was used as the sample to be tested, and the content of β2-MG in the sample to be tested was detected according ...

Embodiment 3

[0139] Embodiment 3, the detection of actual sample disease marker

[0140] Take 5 samples of serum from healthy subjects (informed consent of the examiners) as the samples to be tested, which are respectively recorded as sample 1, sample 2, sample 3, sample 4 and sample 5. The content of the β2-MG marker in the sample to be tested was detected by using the kit and its detection method in Example 1. According to the rare earth metal obtained 151 Eu and 153 Eu signal strength and figure 2 The concentration of β2-MG in each sample was obtained separately from the standard curve shown. The concentrations of β2-MG in sample 1, sample 2, sample 3, sample 4 and sample 5 were 1.21 μg / mL, 0.72 μg / mL, 0.48 μg / mL, 0.91 μg / mL and 1.34 μg / mL, respectively.

[0141] In order to verify the accuracy of this method, commercialized kits (β2-microglobulin detection kits, purchased from Wuxi Jiangyuan Industrial Technology Trade Corporation) were used to measure the serum levels of the abov...

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Abstract

The invention discloses an internal standard correction-based immune precise quantitative analysis method and a special kit thereof. According to the method, an internal standard probe is added in animmune reaction process, simultaneous detection of multiple components can be performed by using a plasma mass spectrometer; and accurate quantitative analysis of immunity is realized by correcting random errors in the immune reaction process and an instrument detection process through the internal standard probe. The internal standard probe strategy designed by the method of the invention not only can be used for competitive immunoassay, but also can be used for non-competitive immunoassay. Therefore, the internal standard correction-based accurate immunoassay method can be used for preparingimmunoassay kits for all clinical disease markers. The method is expected to play a positive role in the fields of clinical detection, individualized treatment, precision medicine and the like.

Description

technical field [0001] The invention belongs to the technical field of biological detection and immune analysis, and in particular relates to an accurate quantitative analysis method of immunity based on internal standard correction and plasma mass spectrometry detection and a special kit thereof. Background technique [0002] Antibody-based immune response is the most basic and common method for specific object detection. The radioimmunoassay (RIA), which was first developed in the 1960s, created a new chapter in the quantitative detection of specific biomolecules. Due to the limitations of labeled radioactive elements such as radioactive hazards, non-radioactive immunoassay methods, such as enzyme-linked immunoassay (ELISA) and chemiluminescence immunoassay (CLIA), have been gradually developed, and have been widely used in life science research and clinical testing. field. However, in the current conventional immunoassay technology, it is difficult to realize the simult...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/534G01N33/68G01N33/60G01N33/543G01N27/62
CPCG01N33/534G01N33/60G01N33/54326G01N33/543G01N33/6803G01N27/626
Inventor 张新荣孙公伟邢志张四纯张钰清胡志安
Owner TSINGHUA UNIV
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