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Thymidine kinase chemiluminescent detection kit and preparation method thereof

A thymidine kinase chemistry, detection kit technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc., can solve the problem that the accuracy of detection results cannot be guaranteed, impurity control is not strict, sensitivity It can improve the detection ability, stable experimental results, and simple and convenient operation.

Pending Publication Date: 2019-10-18
安徽恩禾生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a thymidine kinase chemiluminescence detection kit, which solves the problems of long detection time, low sensitivity and poor repeatability of existing detection methods, and the control of impurities is not strict, and the accuracy of detection results cannot be guaranteed. The problem

Method used

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  • Thymidine kinase chemiluminescent detection kit and preparation method thereof
  • Thymidine kinase chemiluminescent detection kit and preparation method thereof
  • Thymidine kinase chemiluminescent detection kit and preparation method thereof

Examples

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Effect test

Embodiment 1

[0036] A thymidine kinase chemiluminescence detection kit, the kit adopts the double antibody sandwich method to detect the content of TK1 in human serum, the kit includes a public liquid, a coating carrier and a working solution; the public liquid includes a chemiluminescent substrate, a stop solution and a concentrated Washing solution, carrier adopts microwell plate, working solution includes enzyme conjugate, conjugated antibody, quality inspection product and calibration product; among them, enzyme conjugate uses horseradish peroxidase as labeled enzyme, and chemiluminescence substrate adopts DAB chromogenic solution , DAB chromogenic solution includes chromogenic solution A and chromogenic solution B.

Embodiment 2

[0038] The specific preparation method of thymidine kinase chemiluminescence detection kit in embodiment 1 comprises the following steps:

[0039] (1) Prepare coating carrier; anti-TK1 antibody is diluted with coating buffer, take the microplate to be coated, load the diluted anti-TK1 antibody on the microplate, after coating, absorb the coating buffer Then add blocking solution, place at 37°C for 3 hours or overnight at room temperature; discard the blocking solution, dry the coated carrier, add a desiccant to seal the package, and obtain the anti-TK1 antibody-coated carrier.

[0040] (2) Prepare public liquid:

[0041] A, prepare stop solution: stop solution is the hematoxylin staining solution of pH7.2, 6mmol / L;

[0042] B, prepare DAB color developing solution; Color developing solution A is the Tris-HCl solution of pH7.2, 10mmol / L, and color developing solution B is the hydrogen peroxide solution of pH6.0, 10mmol / L;

[0043] C, prepare concentrated washing solution; con...

Embodiment 3

[0049] The specific detection method using the thymidine kinase chemiluminescence detection kit in Examples 1 and 2:

[0050] Including the following steps:

[0051] 1) Add the sample to be tested, 100 μL / well, and incubate at 37°C for 60 minutes;

[0052] 2) Add enzyme: add two drops or 100ul of enzyme marker to each well;

[0053] 3) Incubation: seal the strips with parafilm (to prevent contamination), and incubate at 37°C for 60 minutes;

[0054] 4) Plate washing: take out the reaction plate, shake off all the liquid in the plate, wash the plate 5 times, and pat dry. Air bubbles should be avoided during the washing process;

[0055]5) Add one drop or 50ul of chromogen A and B solutions to each well, and mix well;

[0056] 6) Determination: Use a semi-automatic luminometer to measure the OD. value of each well;

[0057] 7) Calculation of results: take the concentration of the calibrator as the abscissa, and the OD. value as the ordinate. The concentration can be found o...

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Abstract

The invention relates to the technical field of thymidine kinase detection, and particularly discloses a thymidine kinase chemiluminescent detection kit. The kit is characterized in that the content of TK1 in human serum is determined by a double antibody sandwich method. The kit comprises public liquid, a coated carrier and working liquid, wherein the public liquid is prepared from a chemiluminescent substrate, a stop solution and concentrated washing liquid; the carrier is a microplate; and the working liquid is prepared from an enzyme conjugate, a binding antibody, a quality testing productand a calibration product. Through adoption of the kit, the defects in the prior art are overcome. A chemiluminescent method is used for detecting, so that the detection period is controlled within 50 minutes; the operation is easy and convenient; the sensitivity is improved greatly; and a test box has a stable experimental result and high repeatability.

Description

technical field [0001] The invention relates to the technical field of thymidine kinase detection, in particular to a thymidine kinase chemiluminescence detection kit and a preparation method thereof. Background technique [0002] Thymidine kinase (TK1) is a kinase involved in DNA synthesis. Thymidine kinase (TK1) is considered to be a new tumor marker with more reference value, and its increase or decrease is closely related to the occurrence, development and metastasis of tumors. Both the TK1 enzyme activity and its corresponding protein content rapidly increased and accumulated in the early S phase, and reached the highest level in mitotic cells. [0003] The existing detection methods have low detection rate, poor specificity, and are prone to false positive and false negative problems, which greatly affects the evaluation of the patient's condition. Therefore, it is necessary to improve the shortcomings of traditional detection methods through appropriate detection an...

Claims

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Application Information

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IPC IPC(8): G01N33/573G01N33/543G01N33/535G01N21/76
CPCG01N33/573G01N33/543G01N33/535G01N21/76
Inventor 余旭亮
Owner 安徽恩禾生物技术有限公司