Kit for determining ornithine decarboxylase by using marker immunological technique and its determination method

A technology of ornithine decarboxylase and kit, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of radioactive pollution, low sensitivity, complicated operation, etc., and achieve simple operation, no pollution, and high sensitivity Effect

Inactive Publication Date: 2002-11-20
SHANDONG MEDICAL BIO TECH RES CENT
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  • Abstract
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  • Application Information

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Problems solved by technology

[0003] The present invention provides a kit for measuring ornithine decarboxylase by using marker immunoassay technology and its assay method, which indirectly reflects the activity of ornithine decarboxylase by measuring the antigen content of ornithine decarboxylase by using ma

Method used

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Examples

Experimental program
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Embodiment 1

[0070] Detect ornithine decarboxylase in tissue extracts, plasma, serum or other tissue fluids. Take the required amount of microwell strips coated with anti-ornithine decarboxylase antibody, add 50ul tissue extract, and make standard 0, 10, 20, 40, 100 ng / ml and negative control at the same time, add biological to each well 50ul of labeled anti-ornithine decarboxylase antibody, incubate at 37°C for 1 hour, wash 5 times, add 100ul horseradish peroxidase-labeled streptavidin to each well, incubate at 37°C for 40 minutes, wash , Add 50ul each of the developer A and B to each well. After avoiding light at 37°C for 10-15 minutes, add 50ul stop solution to each well, and test at 450nm on the microplate reader. Draw a standard curve with the OD value of each standard well and the corresponding concentration, and check the corresponding concentration from the standard curve according to the OD value of the sample well.

Embodiment 2

[0072] Determination of ornithine decarboxylase in cells (drug sensitivity test). Take 4ml of the patient’s venous blood, use heparin to prevent coagulation, separate lymphocytes with lymphocyte separation solution, and then count and adjust the number of cells to 107 / ml, 100ul per hole, and contain 2ug / ml PHA and different concentrations of anti-cancer drugs such as: Cyclophosphamide 0,10 -3 , 10 -7 , 10 -9 , Vincristine 0,10 -5 , 10 -7 , 10 -9 , Make a control hole at the same time, put 5% CO 2 Incubate in an incubator at 37°C for 18-24 hours, centrifuge and wash 3 times, then routinely fix and block, add 100ul biotin-labeled anti-ornithine decarboxylase antibody to each well, 37°C for 1-3 hours, wash 5 times , Add 100ul of horseradish peroxidase-labeled avidin to each well, wash 5 times at 37℃ for 30-60 minutes, add 50ul each of chromogenic reagent A and B to each well, 37℃ for 10-15 minutes, microplate reader The OD value is measured at 450nm, and the OD value can be used to d...

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Abstract

The kit for determining ornithine decarboxylase by using labelling immune technology contains the following various reagents: precoated plate coated by antibody for resisting ornithine decarboxylase;biotin labelled antibody for resisting ornithine decarboxylase; horseradish peroxidase labelled avidin; ornithine decarboxylase standard product; sample diluent; washing liquor; development reagent Aand B; and stop buffer. Said invention utilizes the labelling immune assey technology to determine ornithine decarboxlase dantigen content to indirectly reflect the activity of ornithine decarboxylase.

Description

Technical field [0001] The invention relates to the technical field of measuring ornithine decarboxylase. Background technique [0002] Ornithine decarboxylase is widely present in animals and is a marker enzyme of cell proliferation. Experiments have shown that the activity of this enzyme is significantly increased in many tumor tissues. According to statistics from relevant medical circles, the activity of ornithine decarboxylase in esophageal cancer, gastric cancer, colorectal cancer and liver cancer tissues is significantly increased. At present, the medical field has regarded ornithine decarboxylase as a tumor marker, so the determination of its activity is particularly important. The traditional method for determination of ornithine decarboxylase is: 1. After the specimen is collected, it is immediately placed in PH7.2 containing isotope-labeled DL-1-[ 14 C] L-ornithine buffer, and then stored in a low temperature environment, so that ornithine decarboxylase and isotope are...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/535G01N33/573
Inventor 朱有名韩金祥尤瑞·巴克拉克
Owner SHANDONG MEDICAL BIO TECH RES CENT
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