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Quality test method of blood sample containing erythrocyte, and test kit

A quality detection method and technology for blood samples, applied in material excitation analysis, fluorescence/phosphorescence, etc., can solve problems such as failure to popularize, lack of auxiliary quality detection methods, difficulty in evaluating the effectiveness of new methods, etc., and achieve high speed and accuracy. High performance and high sensitivity

Inactive Publication Date: 2015-05-27
李捷
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of auxiliary quality detection methods, it is difficult to evaluate the effectiveness of the new method, so it has not been promoted.

Method used

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  • Quality test method of blood sample containing erythrocyte, and test kit
  • Quality test method of blood sample containing erythrocyte, and test kit
  • Quality test method of blood sample containing erythrocyte, and test kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The preparation of the fluorescence resonance energy transferase substrate is completed by a solid-phase peptide synthesizer, which mainly includes the following steps:

[0042] (1) Immobilization of resin and amino acid: First, cross-link the hydroxyl group of the hydroxyl-terminal amino acid synthesized from the peptide chain to be synthesized (from the C' carboxyl end to the N' amino end) with an insoluble polymer resin in a covalent bond structure polystyrene attached.

[0043] (2) Fluorescence labeling of the first amino acid: Depending on the nature of the fluorophore, different labeling techniques will be used, such as the labeling of the fluorophore EDANS (5-(2-aminoethylamino)-1-naphthalenesulfonic acid) It is connected by a covalent bond with the carboxyl group of the side chain of glutamic acid.

[0044] (3) Protection and deprotection of the amino group, carboxyl group and side chain of amino acid: protect the amino group that does not participate in the el...

Embodiment 2

[0056] Kit including fluorescence resonance energy transfer enzyme substrate, including: blood sample non-cellular component separation device, injection bottle filled with fluorescence resonance energy transfer enzyme substrate, pure water, fluorescent instrument with time setting function and standard curve setting function Detection device, fluorescent standard and instructions for use.

[0057] Blood samples were tested using a kit including a FRET substrate as follows:

[0058] (1) Liquid biological samples to be inspected: Any liquid biological samples containing red blood cells, such as blood samples, can be evaluated at any time using the technology involved in the present invention.

[0059] (2) Separation of non-cellular components of the tested liquid biological sample: The traditional centrifuge separation method can be used to separate the non-cellular components of the tested liquid biological sample, which takes 15 minutes. In order to simplify the separation s...

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Abstract

The invention provides a quality test method of a blood sample containing erythrocytes, wherein the content of protease, which is specific in erythrocytes, in a non-erythrocyte component is employed as a parameter for evaluating the quality of the blood sample. The invention also provides a protease substrate having a fluorescence resonance energy transfer character. The structural formula of the protease substrate is D-L-M, herein the D refers to a fluorescent group, the L refers to a linkage group and the M refers to an energy adsorption group. The linkage group is a peptide chain containing tyrosine. In the invention, by means of detection of protease activity specially belonging to interior of cells in the non-erythrocyte component in a liquid biological sample containing cell components, cell rupture degree in the sample can be obtained. To a fresh sample being free of cell rupture during sampling, by means of the method, damage of the cells during processes of collection, storage and transportation of the sample can be obtained. The method has an important significance on evaluation of quality of a blood bank sample used in blood transfusion.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a method for detecting the quality of blood samples containing red blood cells, and also relates to a protease enzyme substrate with fluorescence resonance energy transfer characteristics used in the method, and also relates to a quality detection method for blood samples containing red blood cells kit. Background technique [0002] After the biological sample is separated from the body, due to the lack of necessary living environment such as oxygen and nutrient supply, it will naturally rot and deteriorate, and the biomolecules inside will degrade, resulting in the loss of biological information and loss of cell function, thereby affecting the usable value of the sample; In clinical work, sample degradation can directly lead to misdiagnosis and mistreatment. [0003] Blood samples containing erythrocytes are important biological samples, and their clinical value ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 李捷
Owner 李捷
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