Chemical luminescence immune assay determination reagent kit for prostate gland acid phosphatase and preparation method thereof

A technology of phosphatase chemistry and acid phosphatase, applied in the field of clinical blood immunoassay, can solve the problems of narrow detection range, inability to store for a long time, short half-life of radionuclides, etc., and achieves easy popularization and application, no radioactive pollution, and reduced inspection costs. Effect

Inactive Publication Date: 2009-02-18
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the technology is mature, advanced and practical. The methods widely used in the quantitative analysis of PAP are mainly radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA). Among them, RIA must use 125 Labeling of radioactive elements such as I, the detection equipment is complicated and expensive, and special instruments must be used for measurement. The half-life of the radionuclide is short and cannot be stored for a long time.
ELISA detection sensitivity is not high enough, the detection range is narrow, there are many influencing factors, it is easy to cause false negatives and false positives, and it cannot meet the clinical requirements.

Method used

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  • Chemical luminescence immune assay determination reagent kit for prostate gland acid phosphatase and preparation method thereof
  • Chemical luminescence immune assay determination reagent kit for prostate gland acid phosphatase and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Preparation of Prostate Acid Phosphatase Chemiluminescence Immunoassay Assay Kit of the present invention

[0044] 1. Preparation of enzyme-labeled prostatic acid phosphatase monoclonal antibody

[0045] 1. Improved glutaraldehyde cross-linking method to label alkaline phosphatase (ALP)

[0046] 1) Take 0.50mg of ALP and 1.0mg of monoclonal antibody, dissolve them in normal saline respectively, mix them, and control the final volume at 0.50ml;

[0047] 2) Add 0.50ml of 1% glutaraldehyde, stir magnetically at room temperature for 15 minutes, and react in the dark for 4 hours;

[0048]3) Add 0.10ml of 1.0mol / L ethanolamine and incubate at room temperature for 2h;

[0049] 4) Under the condition of 4°C, use PBS and magnetic stirring to dialyze overnight, and change the dialysate twice;

[0050] 5) Transfer the dialysis product into a reagent bottle, add an equal volume of glycerin, mix well, add 1% BSA, and store in an airtight freezer for later use.

[0051]...

Embodiment 2～3

[0115] Embodiment 2~3 preparation prostatic acid phosphatase chemiluminescence immunoassay assay kit of the present invention

[0116] The PAP chemiluminescent immunoassay assay kit was prepared in the same manner as in Example 1 except that plastic beads and plastic tubes were used as carriers respectively.

Embodiment 4

[0117] Embodiment 4 The usage method of the kit of the present invention

[0118]Before using this kit for experiments, it is necessary to take out the solid-phase antibody, calibrator / test sample, and labeled antibody solution and place them at room temperature for 15 to 30 minutes to allow them to equilibrate to room temperature; after that, prepare a suitable micro-sampler and corresponding Tips and check whether the chemiluminescence instrument and auxiliary equipment, such as a plate washer, are working properly.

[0119] The specific operation steps of using this kit to carry out the experiment according to the method of embodiment 1 are as follows:

[0120] 1) Place the strips required for the experiment (antibody-coated, 96 / 48 wells can be disassembled into 12 / 6*8*1 wells) on the plate rack;

[0121] 2) Add the sample to be tested and calibrators of each concentration in the reaction wells, add 0, 2, 5, 10, 25, 60 ng / ml of 25 μl of the calibrators to each well, set a ...

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Abstract

The invention provides a chemiluminescence immunoassay test kit of prostatic acid phosphatase, which belongs to the clinical blood detecting and assaying technique field. The test kit includes: 1) prostatic acid phosphatase calibrating article; 2) solid-phase carrier which is coated by monoclonal antibody against prostatic acid phosphatase; 3) monoclonal antibody against prostatic acid phosphatase which is labeled by enzyme; 4) chemiluminescence zymolyte on which the enzyme reacts; 5) washing liquid. Furthermore, the invention also provides a preparing method of the test kit which includes steps as following: the calibrating article is prepared; the solid-phase carrier is coated by the monoclonal antibody against prostatic acid phosphatase; the monoclonal antibody against prostatic acid phosphatase is labeled by the enzyme; the calibrating article, the enzyme labeled antibody and the chemiluminescence zymolyte above can be filled and packed through separate packing; and the finished produce is prepared after assembly. With security and reliability, the test kit has the advantages that the sensitivity is high, and the specificity is strong.

Description

technical field [0001] The invention belongs to the technical field of clinical blood immune detection. Specifically, the invention provides a prostatic acid phosphatase chemiluminescence immunoassay assay kit and a preparation method thereof. The kit of the invention has the advantages of simplicity, speed, sensitivity, stability and the like. The detection kit has high sensitivity and strong specificity. Background technique [0002] Prostate cancer is second only to lung cancer among men who die from cancer in European and American countries. With the development of health inspection in our country, prostate cancer screening has also been put on the agenda. Prostate acid phosphatase is the main source of male serum acid phosphatase. It is an isoenzyme produced only by lysosomes of prostate epithelial cells. It is a glycoprotein composed of two subunits with a relative molecular weight of about 100,000 , mainly distributed in semen and urine, with a low content in serum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/573G01N21/76
Inventor 石根应希堂胡国茂郑金来唐宝军于尚永
Owner CHEMCLIN DIAGNOSTICS CO LTD
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