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Method for detecting content of C peptide in human serum, kit used by method and preparation method thereof

A kit and reagent technology, applied in the field of medical laboratory science, can solve the problems of radioactive pollution and high detection cost, and achieve the effects of low detection cost, no radioactive pollution and convenient operation.

Inactive Publication Date: 2015-07-01
SHANGHAI CHUANZHI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention overcomes the deficiencies in the prior art, and provides a latex-enhanced immune turbidimetric assay kit that is convenient to operate, low in price, and can be used for clinical determination of C-peptide content in human serum, its preparation method and the use of the reagent The method of detecting the content of C-peptide in human serum by using a box to solve the problems of long detection time, high detection cost and radioactive contamination in current clinical application of C-peptide

Method used

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  • Method for detecting content of C peptide in human serum, kit used by method and preparation method thereof
  • Method for detecting content of C peptide in human serum, kit used by method and preparation method thereof
  • Method for detecting content of C peptide in human serum, kit used by method and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] This example provides the first reagent and its preparation method.

[0033] (1) For the main components and concentration of the first reagent, see the following table:

[0034] Tris buffer pH7.5 40mM Polyethylene glycol 6000 4% Magnesium chloride 5% Tween20 1% TritonX-100 0.05% Bovine serum albumin 0.2% EDTA 0.04% Sodium azide 0.01%

[0035] (2) The preparation method of the first reagent: add the above-mentioned components of corresponding concentration in order in order, stir at low speed until all are dissolved, adjust the pH to 7.5, add distilled water to the required volume, and mix at room temperature for about 30 After filtering with 0.45μm pore size filter, store at 2~8℃ after completion. The first reagent is a colorless or slightly yellow transparent liquid.

Embodiment 2

[0037] This example provides the second reagent and its preparation method.

[0038] (1) For the main components and concentration of the second reagent, see the following table:

[0039]

[0040] (2) Preparation method of the second reagent:

[0041] a) Latex activation: Take a proper amount of latex, wash with 10 times the volume of latex 40mM Tris buffer (pH 7.5), then dilute to 4 times the volume with 0.04M Tris buffer, and add 1% latex volume of 1% EDC Evenly shake and activate, the activation time is 30min;

[0042] b) Antibody labeling: Add C-peptide antibody (addition ratio of 2-6mg antibody per ml of latex) to the activated latex, shake slightly and evenly at 37°C for 2 hours, then add half the volume of blocking buffer at room temperature Shake evenly for 2 hours. The main components and concentrations of the blocking buffer are shown in the following table:

[0043] Bovine serum albumin Tris pH7.5 Glycine Tween20 0.2% 40mM 0.5% 1%

[0044] c) Washing and reconstitution...

Embodiment 3

[0049] This example provides the calibrator and its preparation method.

[0050] (1) For the main components and concentrations of the calibrator, see the following table:

[0051] Pure C-peptide protein 20ng / ml Tris buffer pH7.540mM Bovine serum albumin 0.2% Tween20 1% TritonX-100 0.05% Sodium azide 0.01%

[0052] (2) Preparation method of calibrator: add the above-mentioned components of corresponding concentration in order in order, stir at low speed until all are dissolved, adjust the pH to 7.5, add distilled water to the required volume, and mix at room temperature for about 30 minutes. Filter with a 0.45μm pore size filter to prepare the highest concentration calibrator. A series of calibrators can be obtained from the highest concentration calibrator through multiple dilution. The prepared calibrator should be stored at 2~8℃ for short-term storage, and at minus 20℃ for long-term storage. The calibrator is a yellowish transparent liquid.

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Abstract

The invention relates to a latex enhanced immunoturbidimetry kit for measuring the content of C peptide in human serum, a preparation method of the kit and a method for detecting the content of C peptide in human serum by use of the kit. The kit comprises a first reagent, a second reagent and a calibrator, wherein the first reagent contains buffer solution, electrolyte, a surface active agent, a stabilizing agent, a reaction enhancer and a preservative; the second reagent contains buffer solution, electrolyte, a surface active agent, a preservative and anti-human C peptide antibody enveloped sensitization polystyrene latex particles; and the calibrator contains buffer solution, a surface active agent, a stabilizing agent, a preservative and human C peptide protein. Compared with the existing kit used clinically, the kit provided by the invention has strong specificity, high sensitivity and high accuracy, the kit is suitable for biochemical analyzers of common detection mechanism, has short detection time, can be operated conveniently, has low detection cost, has no radioactive contamination, and is suitable for popularization and application clinically.

Description

Technical field [0001] The invention belongs to the field of medical laboratory science, and specifically relates to a kit for clinically determining the content of C peptide in human serum, a preparation method of the kit and a method for detecting the content of C peptide in human serum by using the kit. Background technique [0002] Diabetes is a group of metabolic syndrome characterized by chronic hyperglycemia caused by absolute or relative insufficient insulin secretion. In the past 30 years, the prevalence of diabetes in my country has increased significantly. According to the findings of the Diabetes Branch of the Chinese Medical Association (CDS), from 2007 to 2008, the prevalence rate of diabetes in adults over 20 years old in my country was 9.7%. In addition, 14.82 million adults had symptoms of prediabetes, accounting for approximately 1.55% of the population. In the current clinical diagnostic screening, serum C-peptide and insulin levels have important clinical ref...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/042
Inventor 陈明王妮妍
Owner SHANGHAI CHUANZHI BIOTECH CO LTD
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