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CA50 detecting diagnostic kit and preparation method thereof

A technology of diagnostic kits and test kits, which can be applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of rapid attenuation of detection signal values, low-end detection sensitivity, and inconsistent attenuation speed, and achieve the degree of detection automation without environmental pollution. , The effect of improving the coating effect and high sensitivity

Pending Publication Date: 2019-03-29
SUZHOU SYM BIO LIFESCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The advantage of ELISA method is that it is easy to operate, does not require complicated instruments, and only needs simple training to operate. However, due to the limitation of the platform, its sensitivity is relatively low, the measurement range is relatively narrow, and the general detection window period is relatively low. longer
Chemiluminescent CLIA method has the advantages of high sensitivity, specificity and wide detection range; the disadvantage is that most plate-type CLIA kits still use the same enzymatic reaction as EIA, using horseradish enzyme or alkaline phosphatase to cross-link antibodies. Luminol or adamantane, etc. are used as substrates to detect dynamic luminescence, resulting in rapid attenuation of the detection signal value and inconsistent signal attenuation speeds between high and low concentrations, which limits its low-end detection sensitivity and stability in immunoquantitative analysis. sex and repeatability

Method used

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  • CA50 detecting diagnostic kit and preparation method thereof
  • CA50 detecting diagnostic kit and preparation method thereof
  • CA50 detecting diagnostic kit and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Preparation of a diagnostic kit for detecting CA50

[0031] In this example, using Eu 3+ As a fluorescent marker, the specific operation is as follows:

[0032] 1. Obtaining of coated board:

[0033] The coated reaction plate of the present invention is different from the coated reaction plate in the prior art. A special mold is used in the preparation process. The overall plate frame is made of completely opaque black polystyrene. Through-holes corresponding to the aperture of the board (such as figure 2 shown); the ELISA plate is made of materials with excellent light transmission. Before coating the antibody, it is irradiated with cobalt 60 for 30 days to obtain high adsorption. The adsorption pretreatment coated reaction plate is ready for use, as attached image 3 shown.

[0034] Dilute the anti-CA50 monoclonal antibody to 1-4 μg / mL with carbonate buffer solution, 100 μL / well, wash, seal, and dry after coating overnight, vacuum-seal the microwell st...

Embodiment 2

[0040] Example 2 Detection of Carbohydrate Antigen 50 Using a Detection Kit

[0041] Each plate of the reaction plate is provided with 2 wells of the calibrator (A-F), and 25 μL of the calibrator and the sample to be tested are added to the corresponding wells in sequence, and then 50 μL of the diluent is added to each well. 1 hour reaction time;

[0042] Wash 4 times with washing solution, add 100 μL of europium-labeled working solution to each well, and react for 40 minutes at room temperature (18°C-28°C) on the slow gear of the shaker;

[0043] Wash 6 times with washing solution, add 100 μL enhancement solution to each well, and react for 5 minutes at room temperature (18°C-28°C) on the slow gear of the shaker;

[0044] Place the reaction plate in the fluorescence detector and select the corresponding program to read the value.

Embodiment 3

[0045] Embodiment 3 Comparative experiment between kit of the present invention and enzyme immunodiagnostic kit

[0046] Utilize the time-resolved immunofluorescence diagnostic kit for detecting CA50 of the present invention and the common enzyme-immune diagnostic kit (Beijing North Biology) to detect 205 CA50 samples at the same time, if the initial test results of the samples are inconsistent, then use the two detection kits to test again .

[0047] The test results showed that the test results of 7 samples were inconsistent. Taking the detection results of the ELISA method as a reference, the sensitivity of the time-resolved immunofluorescence method reaches 96.2%, the specificity reaches 97%, and the coincidence rate reaches 96.6% (Table 1).

[0048] Table 1 Comparison of HIV sample results by two detection methods

[0049]

[0050] Comparison of Sensitivity and Specificity

[0051] The above-mentioned 7 samples with inconsistent preliminary test results were tested wi...

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Abstract

The invention relates to a CA50 detecting diagnostic kit and a preparation method thereof. According to the diagnostic kit, an immunoadsorption test principle in a double antibody sandwich method anda time distinguishing immunofluorescence technology are combined to detect the carbohydrate antigen 50 quantitatively; the diagnostic kit can be used to help diagnosis of pancreas, colon, rectal and stomach cancers and monitor the treatment effect; and the diagnostic kit also has the advantages including high specificity, sensitivity, repeatability, stability and detection automatic degree, wide determination scope and free of environment pollution.

Description

technical field [0001] The invention relates to the technical field of tumor and cancer antigen immune analysis, in particular to a diagnostic kit for detecting carbohydrate antigen 50 and a preparation method thereof. Background technique [0002] Carbohydrate antigen (CA50) is a glycoprotein mainly composed of sialic acid lipids and sialic acid glycoproteins, which widely exists in epithelial tissue tumors and is mainly used for auxiliary diagnosis of pancreatic cancer, colon cancer, rectal cancer, and gastric cancer. The increase was most pronounced in patients with pancreatic cancer. CA50 is often used for follow-up monitoring after treatment. When CA50 continues to rise, it indicates that the tumor may remain or metastasize. Ulcerative colitis, liver cirrhosis, melanoma, lymphoma, autoimmune diseases, etc. will also increase CA50. [0003] At present, the main methods for detecting CA50 are: enzyme immunosorbent ELISA method and chemiluminescence CLIA method. The adv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/535G01N33/533
CPCG01N33/533G01N33/535
Inventor 李楠张巍黄加喜
Owner SUZHOU SYM BIO LIFESCI CO LTD
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