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AKR1B10 Chemiluminescence quantitative detection kit and application thereof

A 1B10, chemiluminescent technology, applied in the field of medicine and biology, to achieve the effect of improving sensitivity and specificity, improving sensitivity, and luminescence quickly

Pending Publication Date: 2019-12-17
湖南莱拓福生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is still a lack of effective methods for early detection, curative effect judgment, prognosis evaluation or recurrence monitoring of patients with liver cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer and bladder cancer

Method used

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  • AKR1B10 Chemiluminescence quantitative detection kit and application thereof
  • AKR1B10 Chemiluminescence quantitative detection kit and application thereof
  • AKR1B10 Chemiluminescence quantitative detection kit and application thereof

Examples

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Comparison scheme
Effect test

example 1

[0034] Example 1. Preparation of AKR1B10 Chemiluminescence Quantitative Detection Kit.

[0035] 1. The preparation method of AKR1B10 calibrator and AKR1B10 quality control is as follows:

[0036] Dissolve AKR1B10 antigen with 1x PBS solution to prepare AKR1B10 calibrator, the working concentration is 0, 375, 750, 1500, 3000, 6000pg / ml respectively. The 1x PBS solution has a pH value of 7.2 and contains 0.1% Tween 20 (v / v), 1% bovine serum albumin (w / v), and 0.5% Proclin300 (v / v).

[0037] Dissolve the AKR1B10 antigen in 1x PBS solution to prepare AKR1B10 quality control products, and the working concentrations are 375 and 3000pg / ml respectively. The 1x PBS solution has a pH value of 7.2 and contains 0.1% Tween 20 (v / v), 1% bovine serum albumin (w / v), and 0.5% Proclin300 (v / v).

[0038] Second, the preparation method of biotin-labeled antibody is as follows:

[0039] Biotin-labeled antibody was prepared by coupling rabbit anti-AKR1B10 polyclonal antibody with biotin.

[004...

example 2

[0052] Example 2. Detection of AKR1B10 Antigen Using AKR1B10 Chemiluminescent Quantitative Detection Kit

[0053] The steps to detect AKR1B10 antigen are as follows:

[0054] (1) Take 6 test tubes, add 10ul AKR1B10 calibrator of each component to each tube, then add 100ul biotin-labeled antibody and 100ul acridinium ester-labeled antibody, and shake at 37°C for 15 minutes;

[0055] (2) Add 50ul of streptavidin magnetic bead solution to each tube, shake and react at 37°C for 15 minutes;

[0056] (3) Use magnetic separation, add 300ul cleaning solution to each tube, and wash the reagents in the reaction tube repeatedly for 3 times;

[0057] (4) Add 100ul of excitation solution 1 and excitation solution 2 to each tube, and detect the luminescence intensity with a chemiluminescence instrument;

[0058] (5) Data analysis:

[0059] A standard curve was obtained by four-parameter linear fitting of the concentration and luminescence values ​​of the calibrators (see figure 1 ). Am...

example 3

[0060] Example 3. Using AKR1B10 Chemiluminescent Quantitative Detection Kit to Detect the Content of AKR1B10 Protein in Serum Samples of Normal People and Liver Cancer Patients

[0061] (1) Add 10ul serum samples and AKR1B10 calibrator to each tube respectively, add 100ul biotin-labeled antibody and 100ul acridinium ester-labeled antibody respectively, shake and react at 37°C for 15 minutes;

[0062] (2) Add 50ul of streptavidin magnetic bead solution to each tube, shake and react at 37°C for 15 minutes;

[0063] (3) Use magnetic separation, add 300ul cleaning solution to each tube, and wash the reagents in the reaction tube repeatedly for 3 times;

[0064] (4) Add 100ul of excitation solution 1 and excitation solution 2 to each tube, and detect the luminescence intensity with a chemiluminescence instrument;

[0065] (5) Data analysis:

[0066] Serum samples from 50 healthy people and 105 patients with liver cancer were selected for detection, and the standard curve was obta...

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Abstract

The invention relates to the technical field of medical biology, in particular to a quantitative kit for detecting protein content of Aldo-Keto Reductase 1B10 (AKR1B10) by using a magnetic particle chemiluminescence method, which is used for screening, diagnosing, curative effect judging, prognosis evaluation or relapse monitoring of cancers. The kit comprises an AKR1B10 calibration product, an AKR1B10 quality control product, streptavidin magnetic beads, a biotin labeled antibody, an acridinium ester labeled antibody, an excitation liquid 1 and an excitation liquid 2. The AKR1B10 in serum tobe detected is combined with the biotin labeled antibody and the acridinium ester labeled antibody to form a double-antibody sandwich compound, streptavidin magnetic beads are added into the double-antibody sandwich compound, the excitation liquid 1 and the excitation liquid 2 are used for exciting and detecting luminescence values, and the concentration of AKR1B10 in a sample is calculated by means of a standard curve. The AKR1B10 Chemiluminescence quantitative detection kit adopted for detection has the advantages of good stability, high sensitivity, wide linear range and the like.

Description

technical field [0001] The invention relates to the field of medical biotechnology, in particular to a quantitative kit for detecting the protein content of aldehyde and ketone reductase 1B10 (AKR1B10) by magnetic particle chemiluminescence method, which is used for cancer screening, diagnosis, curative effect judgment and prognosis assessment or monitoring for recurrence. The kit is based on sandwich ELISA immunoturbidimetry, enzyme-linked immunoassay, time-resolved immunofluorescence, chemiluminescence and electrochemiluminescence. Background technique [0002] Liver cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer and bladder cancer are currently multiple and common cancers with a very high mortality rate. At present, there is still a lack of effective methods for early detection, curative effect judgment, prognosis assessment or recurrence monitoring of patients with liver cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer and b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N21/76G01N33/543G01N33/577
CPCG01N33/6893G01N21/76G01N33/54326G01N33/577
Inventor 曹哲陈志勇祝跃球
Owner 湖南莱拓福生物科技有限公司
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