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Reagent kit for early warning and diagnosis of severe hepatitis and hepatic failure and preparation method thereof

A diagnostic kit and early warning technology, applied in measurement devices, instruments, scientific instruments, etc., can solve problems such as unreported explanations

Inactive Publication Date: 2012-02-29
同昕生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report explaining the direct relationship between CXCL10 and severe hepatitis liver failure

Method used

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  • Reagent kit for early warning and diagnosis of severe hepatitis and hepatic failure and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Statistics and verification test of CXCL 10 content in serum of patients with different types of hepatitis.

[0024] Serum samples from three batches of 230 cases provided by You'an Hospital were used, including 106 cases of severe hepatitis (ZG), 36 cases of liver cirrhosis (GY), 38 cases of chronic hepatitis (MG), and 39 cases of normal control serum (D).

[0025] The GraphPad Prism 4 software in the United States was used to test the normality of the variables, and the target expression values ​​between groups were compared using the unpaired T-test of two-sample comparison with a group design and the rank sum test one way ANOVA with Tukey's multiple comparison test, the test level α = 0.05 .

[0026] According to the detection results, the average value and standard error of the expression levels of the target in the four blood samples were calculated.

[0027] Table 1 The average expression of targets in the tested blood samples

[0028] group...

Embodiment 2

[0031] Embodiment 2, preparation and detection method of chemiluminescence immunoassay kit

[0032] 1. Coat the anti-CXCL10 monoclonal antibody to become a chemiluminescence reaction plate.

[0033] ELISA plate is milky white opaque polystyrene 96-well chemiluminescent ELISA plate.

[0034] 0.5ug / ml, 1ug / ml, 2ug / ml, and 4ug / ml monoclonal antibodies were used to coat the microtiter plate, respectively, to detect 0pg / ml and 6000pg / ml standards, and the detection of S5 / S0=6000pg / ml Value and the ratio of the detected value of 0pg / ml, the maximum of the ratio is the optimal coating concentration, and the experimental results are obtained as follows:

[0035] Ratio (10,000 times) / coating concentration

0.5ug / ml

1ug / ml

2ug / ml

4ug / ml

S5 / S0

32

45

70

69

[0036] Therefore, 2ug / ml is selected as the optimal coating concentration, and the optimal coating antibody concentration of the antibody detection plate is 2ug / ml.

[0...

Embodiment 3

[0058] Embodiment 3, kit sensitivity and accuracy test

[0059] 1. Kit sensitivity experiment

[0060] 1. Carry out 20 tests on the zero standard solution, and the average value of the test results plus 3 times the standard deviation is used as the minimum detection limit of the kit.

[0061] Table 1 Zero standard measurement results statistical table pg / mL

[0062] sample number

1

2

3

4

5

6

7

8

measured value

0.05

0.08

0.07

0.07

0.06

0.06

0.05

0.06

sample number

9

10

11

12

13

14

15

16

measured value

0.04

0.06

0.07

0.08

0.05

0.07

0.05

0.08

sample number

17

18

19

20

average

standard deviation

Minimum detection limit

measured value

0.05

0.08

0.04

0.06

0.06

0.01

0.1 ...

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Abstract

The invention relates to a reagent kit for the early warning and the diagnosis of severe hepatitis and hepatic failure and a preparation method thereof, which belong to the medical detection reagent. The reagent kit comprises an enzyme-linked immunosorbnent assay (ELISA) plate which is coated with a CXC chemokine ligand-10 (CXCL10)-resistant monoclonal antibody. The ELISA plate is prepared through the following steps that: (1) the CXCL10 monoclonal antibody is diluted by acetate buffer solution with the pH value of 4.6 until the concentration of the CXCL10 monoclonal antibody is 2 micrograms per milliliter; (2) the CXCL10 monoclonal antibody solution is filled into reaction holes of the blank chemical luminous ELISA plate to be statically placed for 12 hours at the temperature of 4 DEG C; (3) liquid inside the reaction holes is removed, and each reaction hole is adequately washed by washing liquor and is dried; and (4) the ELISA plate is sealed and then is dried. The reagent kit has the advantages that the sensitivity and the accuracy are high, the repeatability is good, the minimum detection limit can be 0.1pg / mL, the variation coefficient range is 6.5%-13.1%, and early warning and diagnosis can be provided for the severe hepatitis and the hepatic failure so as to provide sufficient time for doctors and patients to take reasonable prevention and treatment measures.

Description

technical field [0001] The invention relates to a medical detection reagent, in particular to a kit for early warning and diagnosis of severe hepatitis liver failure and a preparation method thereof. Background technique [0002] The liver is the largest internal organ of the human body and one of the organs with the most complex functions. It is responsible for many very important physiological functions, such as the center of various substance metabolism, secretion of bile, synthesis of proteins and blood coagulation factors, detoxification and other functions. The stability of life and internal environment plays an important role. At present, end-stage liver diseases, such as severe hepatitis, liver failure, liver cirrhosis, and liver cancer, are life-threatening and have extremely high mortality rates, which have always been difficult problems for the world's medical community. According to the report of the World Health Organization, about 2 billion people in the world...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/576G01N33/543G01N21/76
Inventor 吴凡焦守恕李全
Owner 同昕生物技术(北京)有限公司
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