Single particle size based reagent kit capable of simultaneously detecting retionl binding protein in blood serum and urine samples

A technology for binding proteins and urine samples, applied in the field of retinol-binding protein kits, can solve the problems of high production cost and low detection sensitivity, and achieve the effects of low cost, high sensitivity and simple operation

Active Publication Date: 2018-10-09
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 t

Method used

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  • Single particle size based reagent kit capable of simultaneously detecting retionl binding protein in blood serum and urine samples
  • Single particle size based reagent kit capable of simultaneously detecting retionl binding protein in blood serum and urine samples
  • Single particle size based reagent kit capable of simultaneously detecting retionl binding protein in blood serum and urine samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Reagent configuration

[0048] 1.1 Configuration of reagent R1

[0049] In the reagent configuration process, configure the concentration according to the following table. After configuration, pass the 0.22μm filter membrane and place it at 4°C for use.

[0050] Component

Matching one

Matching 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] In the reagent configuration process, configure the reagent R2 buffer according to the concentration in the table below. After configuration, pass the 0.22μm filter membrane and place it at 4°C for standby.

[0054] Component

Matching one

Matching two

Matching three

Hepes / mM

50

80

120

Proclin 300 / %

0.1

0.1

0.1

sucrose / %

1

3

5

NaCl / %

1

2

5

Triton / %

0.1

0.5

1

pH

6.5

7.0

8.5

[0055] 1.2.2 Preparation of reagent R2

[0056] (1) Dilute latex particles with a diameter of 130nm with a p...

Embodiment 2

[0069] Kit detection process

[0070] 2.1 Test conditions:

[0071] Dominant wavelength

600nm

Reaction direction

Positive

temperature reflex

37℃

Reaction method

Two-point finish

[0072] 2.2 Operation process:

[0073]

[0074] The reading point in AU480 is 11-27.

Embodiment 3

[0076] Sensitivity performance evaluation (use the ratio 2 in Example 1 to perform the experiment, and test according to Example 2)

[0077] 3.1 Evaluation of the sensitivity of serum samples

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

5％

4％

[0080] 3.2 Evaluation of the sensitivity of urine samples

[0081] Take 0.9% NaCl as a blank sample, select a low-value urine sample for multiple dilution, prepare samples with different ...

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Abstract

The invention discloses a single particle size based reagent kit capable of simultaneously detecting a retionl binding protein in blood serum and urine samples. The reagent kit comprises a reagent R1,a reagent R2, a calibration product A, and a calibration product B, wherein the reagent R2 comprises emulsion-polyclonal antibody cross-linking product and a reagent R2 buffer solution and the emulsion-polyclonal antibody cross-linking product accounts for 0.05-1.0wt% of the reagent R2 buffer solution; the reagent R2 buffer solution comprises the following ingredients: 50-120mM Hepes buffer solution, 0.1% of Proclin 300, 1-5% of sucrose, 1-5% of NaCl, 0.1-1% of triton and the balance of purified water; and a pH (potential of hydrogen) of the reagent R2 buffer solution is 6.5-8.5. The reagentkit has the advantages that the reagent kit detects blood and urine simultaneously, and is simple to operate, high in sensitivity, wide in linear range, and low in cost (only a single particle size and a polyclonal antibody can meet detection requirements, and an emulsion-polyclonal antibody can meet marking requirements only by single centrifugation during preparation), and market popularizationis facilitated. At the same time, the detection reagent kit is compatible with most biochemical detection equipment on a market and can be put on the market directly.

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 the blood. RBP is mainly synthesized by liver cells and is widely distributed in human serum, cerebrospinal fluid, urine and other body fluids. In the blood, RBP, retinol and prealbumin exist as a 1:1:1 (mol) complex, transporting 90% of the retinol in the body to the body tissues, when RBP binds to the RBP receptor on the cell surface When retinol enters the cell, the complex is disintegrated, and free RBP is filtered out from the glomerulus, most of which are reabsorbed by the proximal tubular epithelial cells and decomposed. [0003] Studies have shown that retinol binding protein is of great significance in the diagnosis of hypertension and early diabetic...

Claims

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Application Information

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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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