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Kit and detection method for detecting thymidine kinase 1 in whole blood

A thymidine kinase and kit technology, applied in the field of immunoassay, can solve the problems of long detection time, inefficient operation, and low detection efficiency of TK1 content in human blood, etc., to achieve rapid and effective removal, efficient operation, and short total detection time Effect

Pending Publication Date: 2022-05-10
BEIJING HOMA BIOLOGICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] When the above-mentioned method is used for the determination of TK1 content in human blood, the requirements for the test sample are relatively high. It is necessary to perform centrifugation on the obtained human whole blood and then obtain a serum sample for detection. Then, the above-mentioned detection kit and When the method is used in medical testing, the detection time is too long after the human whole blood sample is obtained, which makes the detection efficiency of TK1 content in human blood low; in addition, when preparing the above detection kit, it is often necessary to add an antibody package Plates, magnetic particles and other antibody carriers are used to coat antibodies, and the carrier needs to be washed during the detection process. These steps make the detection process complicated and inefficient.

Method used

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  • Kit and detection method for detecting thymidine kinase 1 in whole blood
  • Kit and detection method for detecting thymidine kinase 1 in whole blood

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preparation example Construction

[0048] Preparation of magnetic separation reagents

[0049] The preparation method of the magnetic separation reagent of the present application comprises the following steps: take carboxyl magnetic beads, wash with buffer solution 1 for several times and then set aside; then add EDC reagent to activate the carboxyl magnetic beads, mix them uniformly and set aside; prepare the above solution with buffer Liquid 1 was washed to obtain the magnetic separation reagent.

[0050] Wherein, the size of the carboxyl magnetic beads can be 2-5um, such as 3um, 4um. Due to the small particle size of the carboxyl magnetic beads, when the carboxyl magnetic beads are placed in the container, they are in a fluid state. Therefore, in the expression of this application, the amount of the relevant solution is calculated by the volume of the carboxyl magnetic beads.

[0051] Buffer 1 is an acidic buffer, which may be MES buffer at pH 6.0.

[0052] Based on the monomer volume of carboxyl magnetic...

preparation example

[0090] Preparation Example of Magnetic Separation Reagent

[0091] The preparation method of magnetic separation reagent specifically comprises the following steps:

[0092]Take 50uL of carboxyl magnetic beads and wash them with buffer 1. The buffer selected in this preparation is MES buffer at pH 6.0. Wash the carboxyl magnetic beads 3 times, adding 2 mL of MES buffer each time, discard Carboxyl magnetic beads are reserved after the washing solution for future use. Weigh 5 mg of EDC reagent and add it to 1 mL of MES solution at pH 6.0 to obtain EDC reagent-MES solution. The content of EDC in EDC reagent-MES solution is 5 mg / mL, mix well and set aside.

[0093] All the EDC reagent-MES solution obtained above was added to the spare carboxyl magnetic beads above, so that the carboxyl magnetic beads were activated, and the final solution obtained was the magnetic separation reagent. This activation process makes the carboxyl magnetic beads have activated amino acid sites for bi...

Embodiment

[0140] This embodiment provides a kit for detecting thymidine kinase 1 in whole blood. The kit of the present application includes reagent R1, reagent R2, reagent R3 and reagent R4. The R1 reagent is a mixture of magnetic separation reagent, 1.3 mg / L anti-erythrocyte antibody, tween-20 and BSA-V; the R1 reagent is contained in a reagent bottle, and the volume of the R1 reagent is 6 mL. The R2 reagent is a mixture of 0.02 mg / L horseradish peroxidase-labeled TK1 monoclonal antibody, goat and mouse serum; the R2 reagent is contained in a reagent bottle, and the volume of the R2 reagent is 12 mL. The R3 reagent is a mixture of 0.02 mg / L Acridan-labeled TK1 monoclonal antibody, goat and mouse serum; the R3 reagent is contained in a reagent bottle, and the volume of the R3 reagent is 12 mL. Reagent R4 is physiological saline; R4 reagent is contained in a reagent bottle with a capacity of 20mL.

[0141] The composition of the kit for detecting thymidine kinase 1 in whole blood of th...

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Abstract

The invention discloses a kit for detecting thymidine kinase 1 in whole blood and a detection method, belongs to the technical field of immunodetection, and solves the technical problem of long time consumption of an existing detection method. The kit comprises a reagent R1, a reagent R2 and a reagent R3, wherein the R1 reagent comprises a magnetic separation reagent and an anti-erythrocyte antibody; the R2 reagent comprises a TK1 monoclonal antibody labeled by horse radish peroxidase; the R3 reagent comprises a TK1 monoclonal antibody marked by Acridan; the method comprises the following steps: S1, adding a reagent R4 and a reagent R1 into human whole blood, reacting, and carrying out magnetic separation to obtain a reaction solution A; s2, adding a reagent R2 and a reagent R3 into the reaction solution A, and incubating to obtain a reaction solution B; and S3, adding an excitation liquid separant into the reaction liquid B, and determining. The kit and the method are used for detecting the TK1 content in human whole blood, and have the advantages of high detection efficiency and less time consumption.

Description

technical field [0001] The invention relates to the technical field of immune detection, more specifically, it relates to a kit and a detection method for detecting thymidine kinase 1 in whole blood. Background technique [0002] Thymidine kinase 1 (TK1), also known as cytoplasmic thymidine kinase, is the first serological cell proliferation marker. TK1 is closely related to cell proliferation: DNA replication synthesis is the key to maintaining the consistency of old and new cells, and is an important step in cell proliferation, which occurs in the S phase of cell division; cell proliferation uses four deoxynucleotides as raw materials, and TK1 is Among them, the key enzyme of dTTP synthesis, so TK1 can monitor the speed of abnormal cell proliferation. [0003] The application of TK1 mainly has two aspects, (1) physical examination application: to assess the risk of malignant transformation of various proliferative diseases, and proliferative diseases in excessive prolifer...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/531G01N33/58G01N33/577G01N33/537G01N21/76
CPCG01N33/573G01N33/531G01N33/581G01N33/577G01N33/537G01N21/76G01N2333/9121
Inventor 赵科
Owner BEIJING HOMA BIOLOGICAL ENG
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