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APOE kit based on catalytic signal amplification

A signal amplification and kit technology, applied in the field of medical immunology in vitro diagnosis, can solve the problems of high price, large batch-to-batch difference, and low sensitivity, and achieve the effects of reducing usage, improving sensitivity, and reducing costs

Active Publication Date: 2018-05-18
SHANGHAI JIEMEN BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the immunoturbidimetric method is widely used in the domestic detection of ApoE. Although this method is easy to operate, its sensitivity is not high, and it cannot meet the clinical requirements for ApoE quantification.
The reason is that the antibodies used in domestic ApoE detection reagents have problems such as low purity and large batch-to-batch variation, and the main source of antigens mainly depends on imports, which are expensive

Method used

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  • APOE kit based on catalytic signal amplification
  • APOE kit based on catalytic signal amplification
  • APOE kit based on catalytic signal amplification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The present embodiment provides a kind of ApoE detection kit based on signal amplification, which is composed as follows:

[0032] 1. Reagent R1 is:

[0033]

[0034] 2. Reagent R2 is:

[0035]

[0036] 3. Calibrator

[0037] 4 bovine serum matrix calibrators with ApoE concentrations of 1.25mg / dl, 2.5mg / dl, 5mg / dl and 10mg / dl; each bovine serum matrix calibrator also includes glycerol 1%, Proclin300 0.5%, Tween -20 1%.

[0038] The preparation of the bovine serum matrix calibrator:

[0039] Add 1% glycerol, 0.5% Proclin300, and 1% Tween-20 to the processed bovine serum to obtain a calibration product dilution. ApoE was dissolved in the prepared solution similar to human serum matrix to prepare different concentrations of calibrator (1.25mg / dl, 2.5mg / dl, 5mg / dl, 10mg / dl).

[0040] The ApoE detection kit described in this example is applicable to various types of automatic biochemical analyzers. Taking Hitachi 7170 automatic biochemical analyzer as an example, ...

Embodiment 2

[0045] The present embodiment provides a kind of ApoE detection kit based on signal amplification, which is composed as follows:

[0046] 1. Reagent R1 is:

[0047]

[0048] 2. Reagent R2 is:

[0049]

[0050] 3. Calibrator

[0051] 4 bovine serum matrix calibrators with ApoE concentrations of 1.25mg / dl, 2.5mg / dl, 5mg / dl and 10mg / dl; each bovine serum matrix calibrator also includes glycerin 2.5%, Proclin300 1.5%, Tween -20 8%.

[0052] The preparation of the bovine serum matrix calibrator:

[0053] Add 2.5% glycerol, 1.5% Proclin300, and 8% Tween-20 to the processed bovine serum to obtain a calibration product dilution. ApoE was dissolved in the prepared solution similar to human serum matrix to prepare different concentrations of calibrator (1.25mg / dl, 2.5mg / dl, 5mg / dl, 10mg / dl).

[0054] The specific operation method of the kit is the same as in Example 1.

Embodiment 3

[0056] The present embodiment provides a kind of ApoE detection kit based on signal amplification, which is composed as follows:

[0057] 1. Reagent R1 is:

[0058]

[0059] 2. Reagent R2 is:

[0060]

[0061] 3. Calibrator

[0062] 4 bovine serum matrix calibrators with ApoE concentrations of 1.25mg / dl, 2.5mg / dl, 5mg / dl and 10mg / dl; each bovine serum matrix calibrator also includes glycerol 3%, Proclin300 2.2%, Tween -20 10%.

[0063] The preparation of the bovine serum matrix calibrator:

[0064] Add 3% glycerol, 2.2% Proclin300, and 10% Tween-20 to the processed bovine serum to obtain a calibration product dilution. ApoE was dissolved in the prepared solution similar to human serum matrix to prepare different concentrations of calibrator (1.25mg / dl, 2.5mg / dl, 5mg / dl, 10mg / dl).

[0065] The specific operation method of the kit is the same as in Example 1.

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Abstract

The invention provides an ApoE kit based on catalyzed signal amplifying. The kit comprises a reagent R1, a reagent R2 and a calibrator; and the reagent R1 is a phosphate buffer system which contains horseradish peroxidase labeled streptavidin, the reagent R2 is a phosphoric acid buffer system which contains -ApoE antibody and biotin labeled bioamine, and the calibrator comprises four bovine serum matrix calibrators with different ApoE concentrations. A low-molecule nitrogen-containing bioamine catalyzed signal amplifying technology is adopted, the bioamine which is quite high in sensitivity reacts with peroxidase, the reaction turbidity of the reagents is improved, the sensitivity of the reagents is increased, the use amount of the antibody is reduced, the costs of the reagents are greatly reduced, the problem that ApoE reagents are not high in detection sensitivity and expensive in price and the like in markets at present is solved, and the sensitivity of a measurement result is improved obviously.

Description

technical field [0001] The invention belongs to the field of in vitro diagnosis of medical immunity, and in particular relates to an ApoE kit based on catalytic signal amplification. Background technique [0002] Apolipoprotein E (ApoE) is a glycoprotein of 299 amino acids bound to phospholipids. Its molecular weight is 34kD. ApoE can be synthesized in various tissues, but the liver is the main one. First, a propeptide containing 317 amino acids is synthesized. Mature ApoE is formed by ylation and asialolation of extracellular fluid; ApoE is secreted into the blood and transferred to lipoproteins. ApoE is a necessary structural protein for lipoproteins. It can participate in lipoprotein metabolism, regulate cholesterol levels, activate lecithin cholesterol acyltransferase (LCAT) by binding to the receptors of various tissue cells, and has certain immune regulation effects. Determination of ApoE provides an important theoretical basis for the diagnosis and treatment of hyp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/323
Inventor 李伟奇陈瑛房君江张秀文林清玉
Owner SHANGHAI JIEMEN BIO TECH