Chemical luminescence immune assay determination reagent kit for gastrin releasing peptide precursor

A gastrin-releasing peptide and chemiluminescence immunology technology, applied in the field of immunoassay medicine, can solve the problems of limited development and use, low analytical sensitivity, etc., and achieve the effects of reducing dosage, improving linear range and enhancing stability

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

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Problems solved by technology

ELISA technology has limited its development

Method used

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  • Chemical luminescence immune assay determination reagent kit for gastrin releasing peptide precursor
  • Chemical luminescence immune assay determination reagent kit for gastrin releasing peptide precursor
  • Chemical luminescence immune assay determination reagent kit for gastrin releasing peptide precursor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation of the gastrin releasing peptide precursor (31-98) chemical method photoimmunoassay assay kit of the present invention

[0024] 1. Preparation of standard products

[0025] Select adult bovine serum as the standard matrix solution to dilute the pure product of proGRP (31-98), the concentrations are 0pg / ml, 30pg / ml, 150pg / ml, 300pg / ml, 600pg / ml, 1200pg / ml;

[0026] 2. Preparation of enzyme markers and determination of enzyme dilution concentration

[0027] 1. The improved glutaraldehyde cross-linking method marks horseradish peroxidase (HRP), and its steps are as follows:

[0028] a. Dissolve 12mg of HRP in 1ml of PBS (0.1Mol / L pH6.8) containing 5mg of proGRP monoclonal antibody, add 4mL of 1% glutaraldehyde solution under slow stirring, and leave at room temperature for 3 hours.

[0029] b. Add 0.1 mL of 0.2 mol / L lysine (0.29 g dissolved in 10 mL of distilled water), and place at 4°C for 2 hours to block the residual aldehyde groups and terminat...

Embodiment 2

[0070] Example 2 Method of using the gastrin releasing peptide precursor chemical method photoimmunoassay assay kit of the present invention

[0071] 1. Sample pretreatment

[0072] Take human fasting serum, centrifuge at 2000-4000rpm for 10min, and take the supernatant, which can be analyzed without any other special treatment.

[0073] 2. Detection steps

[0074] Before using this kit for detection, it is necessary to take out the components packaged 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; adjust the incubator or water bath to 37°C; Prepare a suitable micro-injector and corresponding tips and check whether the chemiluminescence instrument is working normally.

[0075] The specific operation steps for using this kit are as follows:

[0076] 1) Take out the kit and equilibrate to room temperature;

[0077] 2) Add 25 μL of serum sample or serial...

Embodiment 3

[0085] Embodiment 3 The methodological index of the kit of the present invention

[0086] 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:

[0087] 1. Kit sensitivity

[0088] 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 1.37pg / ml.

[0089] 2. Kit precision

[0090] Three batches of the kit prepared in Example 1 were taken respectively for precision experiment. With the kit extracted in Example 1, three high\middle\low value samples of 110pg / mL and 220pg / mL proGRP samples were measured 8 times eac...

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Abstract

The invention relates to the immunoassay medical field, in particularly provides a chemiluminescence immunoassay test kit and a preparing method thereof for pro gastrin releasing peptide (31-98). Through adopting Biotin-Streptoavidin system coated antibody, the invention can improve antibody coating efficiency, and can improve the sensitivity at the same time. The test kit provides diagnosis of small-cell lung cancer with accurate chemiluminescence test kit which has simple operation and sensitive result detecting, so as to meet the requirements in clinical diagnosis.

Description

technical field [0001] The present invention relates to the field of immunoassay medicine. Specifically, the present invention provides a chemiluminescent immunoassay kit for gastrin releasing peptide precursor (31-98) (proGRP(31-98)) and a preparation method thereof. The inventive method adopts a one-step immunoassay method, and the proGRP (31-98) monoclonal antibody is coated with a streptavidin-biotin system. The kit of the invention has the characteristics of simple operation, reliable performance, sensitivity and accuracy, and provides a reliable detection method for the early diagnosis of small cell lung cancer. Background technique [0002] In industrialized societies, the morbidity and mortality of lung cancer rank first among malignant tumors. The main reason is that in areas with developed industries and transportation, the harmful substances that can cause lung cancer, such as benzopyrene carcinogenic hydrocarbons, which are produced after burning oil and coal, p...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/574G01N21/76
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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