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Chitosanase 3-like protein 1 determination kit

A technology of chitinase and kit, which is applied in the field of biological analysis, can solve the problems of low sensitivity and poor specificity of detection methods, and achieve the effects of improving sensitivity, easy operation, and reducing detection costs

Pending Publication Date: 2021-03-19
BEIJING 3S CENTURY TECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to at least solve one of the technical problems existing in the prior art, provide a kind of test kit that utilizes latex immune turbidity method to quantitatively measure chitinase 3-like protein 1 (CHI3L1) content in human serum in vitro, to solve The detection method in the prior art has the defects of low sensitivity and poor specificity

Method used

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  • Chitosanase 3-like protein 1 determination kit
  • Chitosanase 3-like protein 1 determination kit
  • Chitosanase 3-like protein 1 determination kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The preparation of embodiment 1 kit

[0038] Table 1 Kits

[0039]

[0040]

[0041] 1. Preparation of R1 and R2

[0042] (1) Prepare reagent R1 according to Table 1.

[0043] (2) The preparation of reagent R2 is prepared according to the following method:

[0044] Step 1. Add 250 μl of latex microspheres with a particle size of 150 to 300 nm into 1 mL of the first reaction buffer, mix well, add 4 mL of the second reaction buffer, and mix at 28° C. to obtain a latex microsphere solution; this implementation In the example, the specific preparation method of the first reaction buffer and the second reaction buffer is as follows:

[0045] First reaction buffer: pH 6.0-6.5

[0046] MES·H2O 4.888g / L

[0047] NaOH 0.39g / L

[0048] Second reaction buffer: pH 7.2-7.3

[0049] Boric acid 3.0915g / L

[0050] Sodium tetraborate 0.8586g / L.

[0051] Step 2. Add 250 μl of CHI3L1 monoclonal antibody with a concentration of 10 mg / ml to the reaction system in step 1;

[...

Embodiment 2

[0064] The detection method of embodiment 2 kit

[0065] 1. Reagent preparation

[0066] Liquid dual reagents are ready to use right out of the bottle. Store the reagent in the dark at 2-8°C after opening the bottle, and it is stable for 30 days. Calibration products and quality control products need to be reconstituted with 1.00mL of purified water before use, and placed at 10-30°C for 30 minutes to completely dissolve the lyophilized product, then gently mix them and use them as the original calibration product solution and quality control product solution . Dilute the original calibrator solution with purified water as shown in the table below (volume ratio), and dilute into five concentration gradients. The dilution method is as follows:

[0067] Table 2 Original Calibrator Dilution Ratio

[0068] serial number 1 2 3 4 5 Purified water: stock solution 1:0 7:1 3:1 1:1 0:1

[0069] 2. Parameter setting

[0070] Test method: End point method...

Embodiment 3

[0098] Embodiment 3 Carry out the detection of indexes such as linear range to the kit of embodiment 1

[0099] 1. Linear range

[0100] Standard samples were prepared, and the samples of each concentration were measured 3 times with the kit in Example 1 to get the average value. Correlation analysis is carried out on the measured values, such as figure 1 As shown, R=0.9999 and P<0.05. The analytical measurement range of CHI3L1 verified in this experiment is 0-500.50 U / mL, and the results are shown in Table 3.

[0101] Table 3 Test results

[0102]

[0103] 2. Precision

[0104] Intra-assay precision and inter-day precision were determined using the reagents prepared in Example 1. Select low-value (63.04) and high-value (199.25) two levels of quality control serum as evaluation samples, intra-batch precision: repeat the measurement 20 times in a short period of time and under stable conditions, record each measurement Results and calculation of mean (Mean), standard d...

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Abstract

The invention discloses a chitosanase 3-like protein 1 determination kit, and relates to the technical field of biological analysis. The chitosanase 3-like protein 1 determination kit comprises a reagent R1 and a reagent R2; the reagent R1 comprises 50 to 300 mmol / L of sodium citrate, 0.05 to 0.5 mol / L of sodium chloride and 0.02 to 5% of a preservative, and the reagent R2 comprises 50 to 300 mmol / L of sodium citrate, 0.5 to 5% of bovine serum albumin, 0.2 to 2% of Tween 20, 0.02 to 5% of a preservative and 0.5 to 5% of latex particles coated with an anti-chitosanase 3-like protein 1 antibody.The kit provided by the invention has the advantages of high sensitivity, strong specificity, simple operation and the like, can be applied to various full-automatic biochemical analyzers for large-batch determination, is suitable for clinical popularization, and reduces the hospital detection cost.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, in particular to a high-efficiency assay kit for chitinase 3-like protein 1 (CHI3L1) (latex immunoturbidimetric method). Background technique [0002] Chitinase-3-like 1 (CHI3L1) belongs to the glycosyl hydrolase family 18, and various cells can express and secrete CHI3L1, such as macrophages, neutrophils, chondrocytes, fibroblasts, Tumor cells and airway epithelial cells. CHI3L1 can participate in pathological processes such as inflammation, cell proliferation and differentiation, protection of cell apoptosis, promotion of angiogenesis and extracellular matrix remodeling. The expression level of CHl3L1 in the liver is much higher than that in other tissues, and it is a liver-specific or highly liver-enriched gene. CHl3L1 is a good serological indicator of liver fibrosis to distinguish chronic hepatitis B (CHB) from cirrhosis . [0003] At present, histopathological examination, se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/539
CPCG01N33/573G01N33/539
Inventor 彭勇郝立颖刘洁
Owner BEIJING 3S CENTURY TECH CORP
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