Harmless positive control for enzyme-linked immune diagnosis of hepatitis c

A positive control and immunodiagnostic technology, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as lack of vaccines

Inactive Publication Date: 2015-04-29
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

HCV cannot be cultivated by conventional methods at present, and there is no vaccine or specific drug, so prevention is the main way to control its epidemic

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Harmless positive control for ELISA diagnosis of hepatitis C

[0029] 1. Obtaining mouse anti-HCV monoclonal antibody

[0030] Using HCV virus genetically engineered antigens (including proteins encoded by HCV virus Core, NS3, NS4, NS5, E2 and other genes) as immune antigens, immunize BALB / c mice, and fuse spleen lymphocytes of immunized mice with myeloma cells , and then use the limited dilution method to obtain hybridomas that can secrete erythrocyte monoclonal antibodies. Specific steps are as follows:

[0031] Take 0.2 mg / ml HCV virus genetically engineered antigen, and adopt subcutaneous multi-point injection for initial immunization, 0.1 ml / point, four points in total. Two weeks later, the second immunization was performed with the same dosage as above, intraperitoneal injection. Another 2 weeks later, the third immunization, the same dose as above, intraperitoneal injection (5 to 7 days later, the blood was collected to measure its potency). After a...

Embodiment 2

[0042] Example 2 Hepatitis C Indirect Method ELISA Diagnostic Kit

[0043] Utilize the harmless positive control obtained in Example 1 to prepare the hepatitis C indirect enzyme immunoassay diagnostic kit, the composition is as follows:

[0044] Kit composition

48-well configuration

96-well configuration

save

manual

1 copy

1 copy

Sealing film

2 pieces (48)

2 pieces (96)

sealed bag

1

1

Enzyme-coated plate

1×48

1×96

Store at 2-8°C

negative control

0.5ml×1 bottle

0.5ml×1 bottle

Store at 2-8°C

positive control

0.5ml×1 bottle

0.5ml×1 bottle

Store at 2-8°C

Enzyme label reagent

3ml×1 bottle

6ml×1 bottle

Store at 2-8°C

sample diluent

3ml×1 bottle

6ml×1 bottle

Store at 2-8°C

Chromogen A solution

3ml×1 bottle

6ml×1 bottle

Store at 2-8°C

Chromogen B solution

3ml×1 bottle

6ml×1 b...

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PUM

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Abstract

The invention relates to an enzyme-linked immunosorbent kit, and particularly provides harmless positive control for enzyme-linked immune diagnosis of hepatitis c. The positive control is prepared by an anti-mouse HCV antibody and an anti-mouse IgG antibody in the ratio of (2 to1) to (1 to 32) in a mixing manner. According to the immunology method, the problem of source of a positive control material for enzyme-linked immune diagnosis of the hepatitis c is solved; and the potential risk caused by human serum is avoided.

Description

technical field [0001] The invention relates to an ELISA reagent kit, in particular to a harmless positive control for ELISA diagnosis of hepatitis C and its preparation method and application. Background technique [0002] Hepatitis C virus (hepatitis C) is a disease mainly transmitted by blood, caused by hepatitis C virus (hepatitis C virus, HCV) infection, and is prevalent worldwide, with an infection rate of about 3%. The infection rate of the general population in my country is 3.2%, 3.6% in the north of the Yangtze River, and as high as 3.8% and 4.6% in the south-central and northeast regions, respectively. [0003] The harm of hepatitis C is that the cases of spontaneous recovery after HCV infection are rare, and the chronicity rate can be as high as 60% to 85%. The consequences are chronic inflammation, necrosis and fibrosis of the liver, which can easily transform into end-stage liver diseases such as cirrhosis and hepatocellular carcinoma (HCC), with a poor progno...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/56983G01N33/5767G01N2333/186
Inventor 李银生李勇莉杨杰周薇张东芳胡亚男刘辉
Owner XINXIANG MEDICAL UNIV
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