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A kit based on single particle size and capable of simultaneously detecting retinol-binding protein in serum and urine samples

A technology for binding protein and urine samples, which is applied in the field of retinol binding protein kits, can solve the problems of high production cost and low detection sensitivity, and achieve the effect of low cost, high sensitivity and wide linear range

Active Publication Date: 2019-05-03
宁波海尔施智造有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The technical problem to be solved by the present invention is to solve the low detection sensitivity (can only detect RBP in serum) or the high production cost (can detect RBP in serum and urine simultaneously on the market at present, but the production cost based on the technical scheme of double particle size is relatively low). High) technical defect, provide a kind of kit based on single particle size and can detect retinol binding protein in serum and urine sample simultaneously

Method used

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  • A kit based on single particle size and capable of simultaneously detecting retinol-binding protein in serum and urine samples
  • A kit based on single particle size and capable of simultaneously detecting retinol-binding protein in serum and urine samples
  • A kit based on single particle size and capable of simultaneously detecting retinol-binding protein in serum and urine samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Reagent configuration

[0048] 1.1 Configuration of reagent R1

[0049] During the reagent preparation process, prepare according to the concentration in the table below, pass through a 0.22 μm filter membrane after preparation, and store at 4°C for later use.

[0050] components

Proportion one

Proportion two

Matching three

Hepes / mM

50

80

120

Proclin 300 / %

0.1

0.1

0.1

NaCl / %

1

2

5

Triton / %

0.1

0.5

1

pH

6.5

7.0

8.5

[0051] 1.2 Configuration of reagent R2

[0052] 1.2.1 Configuration of reagent R2 buffer

[0053] During the reagent preparation process, the reagent R2 buffer solution was prepared according to the concentration in the table below. After preparation, it was passed through a 0.22 μm filter membrane and stored at 4°C for later use.

[0054] components

Proportion one

Proportion two

Matching three

Hepes / mM

50

80

120 ...

Embodiment 2

[0069] Kit detection process

[0070] 2.1 Test conditions:

[0071] dominant wavelength

600nm

Reaction direction

Forward

temperature reflex

37℃

Response method

two-point endpoint

[0072] 2.2 Operation process:

[0073]

[0074] The reading points in AU480 are 11-27.

Embodiment 3

[0076] Sensitivity performance evaluation (carry out experiment with proportioning 2 in embodiment 1, and detect according to embodiment 2)

[0077] 3.1 Sensitivity determination and evaluation of serum samples

[0078] Take 0.9% NaCl as a blank sample, select a low-value serum sample for doubling dilution, prepare samples with different concentrations (0, 1, 2, 2.5, 3mg / L), repeat the measurement 10 times for each sample, and calculate Mean and standard deviation, the results are shown in the table below and figure 1 , it can be seen that the detection sensitivity of the kit of the present invention to serum RBP is 1 mg / L.

[0079] Concentration (mg / L)

0

1

2

2.5

3

STDV

0.32

0.09

0.16

0.12

0.12

AVERAGE

0.38

0.91

1.81

2.60

3.28

3SD

0.96

0.28

0.49

0.35

0.37

AVERAGE-3SD

/

0.63

1.31

2.24

2.31

AVERAGE+3SD

0.57

/

/

/

/

cv

83%

10%

9% ...

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Abstract

The invention discloses a kit based on a single particle size and capable of simultaneously detecting retinol-binding protein in serum and urine samples. The kit includes reagent R1, reagent R2, calibrator A, calibrator B, and the reagent R2 includes a latex-polyclonal antibody cross-linked product and a reagent R2 buffer, the latex-polyclonal antibody cross-linked product accounts for 0.05-1.0% by mass of the reagent R2, and the reagent R2 buffer component is: 50-120mM Hepes buffer, 0.1% Proclin 300, 1-5% sucrose, 1-5% NaCl, 0.1-1% Triton, the rest is purified water, pH is 6.5-8.5. The kit of the present invention has the advantages of simultaneous measurement of hematuria, simple operation, high sensitivity, wide linear range, and low cost (not only reflected in the fact that only one particle size and polyclonal antibody can meet the detection requirements, but also in the preparation of latex-multiple When cloning antibodies, only one centrifugation can meet the labeling requirements) and facilitate the advantages of marketing. At the same time, the detection kit is compatible with most biochemical detection equipment on the market and can directly enter the market.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit based on a single particle size and capable of simultaneously detecting retinol-binding proteins in serum and urine samples. Background technique [0002] Retinol Binding Protcin (RBP) is a transporter of retinol (vitamin A) in blood. RBP is mainly synthesized by liver cells and widely distributed in human serum, cerebrospinal fluid, urine and other body fluids. In the blood, RBP exists in the form of a 1:1:1 (mol) complex with retinol and prealbumin, transporting 90% of retinol in the body to body tissues, when RBP binds to the RBP receptor on the cell surface When retinol enters the cell, the complex disintegrates, and free RBP is filtered out from the glomerulus, most of which are reabsorbed by the proximal renal tubular epithelial cells and decomposed. [0003] Studies have shown that retinol binding protein is of great significance in the diagnosis of hypertension and ea...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 柳建敏林威彦裘艳群
Owner 宁波海尔施智造有限公司
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