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Early diagnosis kit based on combined detection of four items of liver cancer and application

A combined detection and kit technology, applied in the field of immunodetection, achieves the effects of good stability, reduced detection time and cost, and easy mixing and combination

Pending Publication Date: 2022-04-29
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

[0007] However, there are more or less problems in the accuracy and reliability of the individual detection accuracy and reliability of the above-mentioned liver cancer serum markers.

Method used

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  • Early diagnosis kit based on combined detection of four items of liver cancer and application
  • Early diagnosis kit based on combined detection of four items of liver cancer and application
  • Early diagnosis kit based on combined detection of four items of liver cancer and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1. Preparation of four combined assay kits for liver cancer.

[0036] 1. The preparation method of biotin-labeled antibody is as follows:

[0037] Take biotin-labeled rabbit anti-AKR1B10 polyclonal antibody as an example:

[0038] (1) Replace the rabbit anti-AKR1B10 polyclonal antibody to be labeled with a desalting column into a 1x PBS buffer.

[0039] (2) Mix the antibody in step (1) with biotin at a concentration of 10mM, use a molecular ratio of 20:1 between biotin and antibody, and react with gentle shaking at room temperature (18-25°C) for 2 hours.

[0040] (3) Replace the labeled antibody solution with a desalting column into a 1x PBS solution to remove free biotin and store at -80°C to prepare the desired biotin-labeled antibody.

[0041] The preparation of other biotin-labeled antibodies can be carried out by referring to the above method.

[0042] 2. The preparation method of acridinium ester-labeled antibody is as follows:

[0043] Take acridinium...

Embodiment 2

[0059] Example 2. Detection steps of the four combined assay kits for liver cancer.

[0060] 1. The detection steps of the first reagent, the second reagent and the fourth reagent are as follows:

[0061] (1) Add an appropriate volume of each sample to three reaction cups, and add corresponding volumes of AKR1B10, AFP and DCP calibrator and quality control products to the reaction cups.

[0062] (2) Add the biotin antibody and acridinium ester antibody of AKR1B10, AFP and DCP into the corresponding cuvettes respectively, and the addition amount is 100uL respectively.

[0063] (3) Incubate the cuvettes at 37°C for 10 minutes, then add 30uL streptavidin magnetic beads to each cuvette, and incubate at 37°C for 20 minutes.

[0064] (4) Adsorb the magnetic beads under the action of magnetic force, absorb the liquid in the cuvette, and wash repeatedly 3 times.

[0065] (5) Under the action of the pre-excitation solution and the excitation solution, the acridinium ester is excited ...

Embodiment 3

[0076] Example 3. The normal reference value of the four joint assay kits for liver cancer.

[0077] Adopt the detection method identical with embodiment 2, select 228 routine healthy normal person samples, determine normal reference value according to 95% percentile (P 95 =(228+1)×95%=217). According to the order of sample concentration, in 228 serum samples, the concentration of AKR1B10 corresponding to P95=217 is 373.5pg / mL, the concentration of DCP is ≤161.45mAU / mL, the concentration of AFP is 8.7ng / mL, and the ratio of AFP-L3% ≤10%, see Table 1 for detailed results, (AFP-L3% is the concentration ratio of AFP-L3 and AFP).

[0078] Table 1. Measured values ​​of serum samples of 228 healthy normal persons

[0079]

[0080]

[0081]

[0082]

[0083]

[0084] Therefore, it was determined that the normal reference value of AKR1B10 was ≤350 pg / mL, the normal reference value of AFP was ≤8.7 ng / mL, the normal reference value of AFP-L3 / AFP was ≤10%, and the normal...

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Abstract

The invention belongs to the technical field of immunodetection, and discloses a combined detection kit based on four liver cancer tumor markers: aldoketoreductase 1B10 (AKR1B10), abnormal prothrombin (DCP), alpha fetoprotein (AFP) and alpha fetoprotein variant (AFP-L3). The kit comprises: streptavidin magnetic beads; labeling AKR1B10, DCP (dicumyl peroxide) and AFP antibodies with biotin; labeling AKR1B10, DCP and AFP antibodies with acridinium ester; an LCA magnetic bead, a cleaning solution and an eluent; and AKR1B10, DCP and AFP calibration products and quality control products. By simultaneously detecting the contents of four tumor markers in serum of a subject, the risk of potential development of liver cancer of the subject is analyzed and judged. According to the invention, the sensitivity and specificity of early diagnosis of liver cancer are improved by combined detection of four tumor markers, and a more reliable basis is provided for clinical diagnosis of liver cancer.

Description

technical field [0001] The invention belongs to the technical field of immunoassay, and in particular relates to liver cancer-specific markers aldehyde and ketone reductase 1B10 (AKR1B10), alpha-fetoprotein (AFP), alpha-fetoprotein heterogeneity (AFP-L3) and abnormal prothrombin (DCP) combined assay kit, which can be used for early screening, diagnosis, curative effect judgment, prognosis evaluation or recurrence monitoring of liver cancer or high-risk groups. Background technique [0002] Primary hepatocellular carcinoma (Hepatocellular Carcinoma, HCC) is one of the common malignant tumors in clinical practice. Its main causes are liver cirrhosis, hepatitis, alcoholic and non-alcoholic fatty liver. These patients are collectively referred to as high-risk liver cancer. crowd. According to the statistics of the World Health Organization, there will be 910,000 new cases of liver cancer and 830,000 deaths worldwide in 2020. The early symptoms of liver cancer are not obvious. ...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/543
CPCG01N33/57438G01N33/57484G01N33/57488G01N33/54326G01N2333/974G01N2333/902
Inventor 曹哲祝跃球
Owner 湖南莱拓福生物科技有限公司
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