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A calibrator stabilizer, a detection kit and a detection method for measuring c-peptide

A detection kit and stabilizer technology, applied in the field of biomedical testing, can solve problems such as short shelf life, slow precipitation, and destruction of antigen structure, and achieve the effects of reducing reaction time, optimizing process flow, and improving utilization rate

Active Publication Date: 2020-07-14
GUANGZHOU JINDE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Disadvantage 1: Photoactivated chemiluminescence requires an antibody coating process. Although it is in a homogeneous system during the reaction, it is not completely homogeneous, and the emission intensity depends on various environmental factors. In different environmental systems, the emission intensity and There is a big difference in the time curve, so various external factors must be strictly controlled. For example, if there is no luminous bead within 200nm near the photosensitive bead, the singlet oxygen will decay to the ground state oxygen and enter the next energy cycle. No light signal is generated; the second disadvantage is that the strength of the light signal of photochemiluminescence depends on the size of the sample to a certain extent, there are many interfering substances in the blood, and the sample size for photochemiluminescence is generally very high , so it is easy to be interfered by interfering substances
The general antigen preservation solution will slowly precipitate at room temperature, resulting in the destruction of the antigen structure and the decrease of the concentration
Especially in the environment above 10°C or the environment of the solution is not good, it will lead to a sharp drop in the antigen concentration, which is irreversible, which makes the calibration test invalid, and the general calibrator stabilizer can only ensure that the calibrator Store at 4°C or even -20°C before opening the bottle for one year or even a shorter shelf life, and it is extremely unstable for repeated freezing and thawing and placing it at room temperature after opening the bottle
In addition, it is impossible to completely guarantee that the calibrator is carried out in an environment of 4°C during use and actual transportation.

Method used

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  • A calibrator stabilizer, a detection kit and a detection method for measuring c-peptide
  • A calibrator stabilizer, a detection kit and a detection method for measuring c-peptide
  • A calibrator stabilizer, a detection kit and a detection method for measuring c-peptide

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] 1. Preparation of calibrator stabilizer

[0075] Bovine serum albumin, trehalose, sucrose, mannitol, glycine, gelatin, edetate disodium, casein, magnesium chloride (proportion 5:3:2:1:2:1:2:2:1 ) was added to deionized water and mixed with a glass rod for 10 minutes until completely dissolved, then filtered. Stand at 37°C for 11 hours to obtain mixture A. Add glycerin to the mixed solution A, and mix it with a glass rod for 34 minutes at room temperature. The volume ratio of the mixed solution A to glycerin is 1:1. Collect the powder after freeze-drying to obtain the calibrator stabilizer.

[0076] 2. Preparation of calibrator

[0077] Phosphate buffer 30mmol / L stabilizer 1w / v% pH 7.4

[0078] The test group can be obtained by adding the calibrator stabilizer to the phosphate buffer matrix solution containing preservatives, and the amount of the calibrator stabilizer added to 1L of the matrix solution is 10g. Take the calibrator matrix s...

Embodiment 2

[0085] Embodiment 2, acridinium ester-labeled anti-C-peptide antibody and horseradish peroxidase-labeled anti-C-peptide antibody concentration selection standard allow

[0086] The acridinium ester-labeled anti-C-peptide antibody and the horseradish peroxidase-labeled anti-C-peptide antibody were mixed at different dilutions by using the square array method. The minimum signal-to-noise ratio was greater than 5 and the highest signal-to-noise ratio was greater than 200. The matching relationship with the lowest cost is preferred.

[0087] The acridinium ester-labeled anti-C-peptide antibody was mixed with horseradish peroxidase-labeled anti-C-peptide antibody at different dilutions of 1 / 500, 1 / 1000, 1 / 2000, 1 / 4000, 1 / 8000, and 1 / 16000 Different dilutions of 1 / 500, 1 / 1000, 1 / 2000, 1 / 4000, 1 / 8000, 1 / 16000 were investigated by the square matrix method. The proportions of the two groups were cross-matched, and the optimal proportion was finally selected as Acridan-labeled anti...

Embodiment 3

[0088] Embodiment 3, the present invention measures the detection kit of C peptide

[0089] Calibrator: pH 7.4

[0090] C-peptide antigen 10ng / mL Phosphate buffer 30mmol / L Calibrator Stabilizer 1w / v%

[0091] Reagent R1: pH 7.4

[0092]

[0093]

[0094] Reagent R2: pH 7.4

[0095] Phosphate buffer 30mmol / L Horseradish peroxidase-conjugated anti-C-peptide antibody 5mg / L BSA 0.01w / v% Mannitol 0.5w / v% PEG20000 0.5w / v%

[0096] Reagent R3:

[0097] ascorbic acid 0.1w / v% bovine serum albumin 0.5w / v% sucrose 0.5w / v%

[0098] Reagent R4:

[0099] hydrogen peroxide 10mmol / L bovine serum albumin 0.5w / v% Trehalose 0.5w / v% Methyl p-hydroxycinnamate 0.1w / v%

[0100] The semi-finished product of the kit was prepared according to the above formula, and assembled into a C-peptide detection kit after passing the performance verification.

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Abstract

The invention belongs to the technical field of biomedical examination, and particularly relates to a calibration product stabilizer, a C peptide determination detection kit and a detection method. The kit comprises a calibration product, a reagent 1, a reagent 2, a reagent R3 and a reagent R4. According to the kit, an acridinium ester-labeled anti-C peptide antibody, an antigen and a horse radishperoxidase-labeled anti-C peptide antibody are utilized to form an antibody-antigen-antibody compound. A triggering agent is added into the compound without a washing process, the compound is continuously detected for a period of time, the peak area is calculated every 0.02-0.05 S, and a dosage-reaction curve is made by using C peptide with known concentration and the calculated peak area; and the content of the C peptide in the sample to be detected is calculated according to the curve. The kit for detecting the C peptide by adopting the spatial proximity chemiluminiscence method provided bythe invention has the advantages of strong calibration product stability, strong reagent anti-interference capability, high accuracy, good specificity and wide linear range, and is suitable for beingused by medical and research institutions at all levels in combination with instrument measurement.

Description

technical field [0001] The invention belongs to the technical field of biomedical testing, and in particular relates to a calibrator stabilizer, a detection kit for measuring C-peptide and a detection method. Background technique [0002] C-peptide is a secretory product of pancreatic β cells, and it has a common precursor with insulin—proinsulin. A molecule of proinsulin is cleaved into a molecule of insulin and a molecule of C-peptide under special action. Therefore, in theory, C-peptide and insulin are equally secreted. The physiological function of free C-peptide in the blood is not very clear. However, C-peptide is not destroyed by the liver, and its half-life is significantly longer than that of insulin. Therefore, the determination of C-peptide level can better reflect the function of β-cell synthesis and release of insulin. [0003] For patients who have been treated with insulin, the insulin antibodies produced in the body can interfere with the determination of in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/535G01N21/76
Inventor 丘传添李民友曾华宁吴新伟赵肃清郑盛武
Owner GUANGZHOU JINDE BIOTECH