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Des-gamma-carboxy-pro-thrombin microplate chemiluminescence immune analysis determination reagent kit and preparing method thereof

A chemiluminescence immunoassay and prothrombin technology, applied in the field of chemiluminescence immunoassay kits and their preparation, can solve the problems affecting the early diagnosis of small liver cancer, low detection sensitivity, large cross-reaction, etc., achieve good market value and improve linearity Range, easy operation

Inactive Publication Date: 2009-03-04
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to provide an antibody purification technology and chemiluminescence immunoassay method, and a kit for quantitative determination of DCP chemiluminescence immunoassay, which solves the non-specific problem of large cross-reaction, and solves the problem of low detection sensitivity that affects small liver cancer Problems with Early Diagnosis

Method used

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  • Des-gamma-carboxy-pro-thrombin microplate chemiluminescence immune analysis determination reagent kit and preparing method thereof
  • Des-gamma-carboxy-pro-thrombin microplate chemiluminescence immune analysis determination reagent kit and preparing method thereof
  • Des-gamma-carboxy-pro-thrombin microplate chemiluminescence immune analysis determination reagent kit and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Preparation of de-γcarboxyprothrombin chemical method photoimmunoassay assay kit of the present invention

[0044] 1. Preparation of standard products

[0045] Horse serum, 50% horse serum (diluted in normal saline), adult bovine serum, 50% adult bovine serum (diluted in normal saline), 3% BSA+0.6% hydrolyzed gelatin buffer (diluted in normal saline) were used for five kinds of matrices Select, preferably the one with the highest sensitivity (in RLU S1 / RLU S0 )express.

[0046] Preparation of point S1: Dissolve 10mAU into 1ml of the above five matrices, mix thoroughly, and set aside.

[0047] Then according to the operation process of the present invention, that is: 50 μl of standard substance and 50 μl of enzyme marker were respectively added to the coated microwell plate, incubated at 37°C for 40 min, washed, and 100 μl of luminescent substrate was added to detect S 0 , S 1 point RLU. The results are shown in Table 1. It can be seen from the results ...

Embodiment 2

[0103] Example 2 Method of using the de-gamma carboxyprothrombin chemical method photoimmunoassay assay kit of the present invention

[0104] 1. Sample pretreatment

[0105] Take human fasting serum, centrifuge at 2000-4000rpm for 10min, and take the supernatant for analysis without any other special treatment.

[0106] 2. Detection steps

[0107] Before using this kit for detection, it is necessary to take out the packaged medicines in the kit first: the coated plate, enzyme markers, and standard products are allowed to stand at room temperature, and then used after equilibrating to room temperature; after that, adjust the incubator or water bath. to 37°C; then, prepare a suitable micro-injector and corresponding tips and check whether the chemiluminescence instrument is working normally.

[0108] The specific operation steps of using this kit to carry out the experiment according to the method of embodiment 1 are as follows:

[0109] 1) Take out the kit and equilibrate to r...

Embodiment 3

[0120] Embodiment 3 The methodological index of the kit of the present invention

[0121] The test kit prepared in Example 1 is tested according to the conventional manufacturing and testing procedures in the art, and the results are as follows:

[0122] 1. Kit sensitivity

[0123] Sensitivity is the concentration that is distinguishable from the zero dose point. Measure 10 zero standard points at the same time to obtain the average value and standard deviation of RLU. Calculate the difference between the average value of RLU and twice the standard deviation, and obtain the corresponding concentration value from the working curve by interpolation. The sensitivity determined in the experiment was 4.37mAU / ml.

[0124] 2. Kit precision

[0125] Precision refers to the reproducibility of the analysis results. Prepare three samples with 8 high / medium / low values, and test each sample at the same time in the same analysis. The ratio of the coefficient of variation to the standard...

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Abstract

The invention relates to the medical field of immunoassay, more specially, the invention provides a chemiluminescent immunoassay detection kit for des-gamma-carboxyand-prothrombin (DCP) microporosity plate and a preparation method thereof, and realizes the rapid, sensitive and high-specificity serological detection of DCP with the one step chemiluminescent immunoassay method. The kit of the invention has the advantages of simplicity, convenience, rapidness, sensitivity, stability and the like, eliminates the interference of the prothrombin analogues and the serum cellulose and the analogues thereof, and has the advantage of high specificity.

Description

technical field [0001] The invention relates to the field of immunoanalysis medicine. Specifically, the invention provides a one-step immunoassay method for fast, highly sensitive, and highly specific chemiluminescent immunoassay kits for measuring degamma carboxyprothrombin (DCP) and a preparation method thereof. Background technique [0002] Cancer has posed a serious threat to human survival and health since its birth. At present, the key to tumor treatment lies in early detection. With the early diagnosis of tumors entering the protein era, humoral proteins represented by blood have become a hot spot in the field of tumor marker research. The discovery of more and more tumor markers has put forward higher requirements for the establishment of new detection technologies for rapid development, high sensitivity and high specificity of tumor markers. [0003] Liver cancer is the fifth most common cancer worldwide and the third deadliest. Primary liver cancer accounts for 8...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/543G01N21/76
Inventor 张倩云应希堂宋胜利胡国茂郑金来于尚永
Owner CHEMCLIN DIAGNOSTICS CO LTD
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