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Bifunctional antibody serving as positive control in detection of hepatitis C virus

A hepatitis C virus, bifunctional antibody technology, applied in measurement devices, anti-animal/human immunoglobulin, instruments, etc., can solve the problem of no vaccine and so on

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

[0030] The bifunctional antibody used as positive control when embodiment 1 is used to detect hepatitis C virus

[0031] 1. Obtaining mouse anti-HCV monoclonal antibody

[0032] HCV virus genetically engineered antigen was used as the immune antigen to immunize BALB / c mice, the spleen lymphocytes of the immunized mice were fused with myeloma cells, and then the hybridoma that could secrete erythrocyte monoclonal antibody was obtained by limiting dilution method. Specific steps are as follows:

[0033] Take 0.2 mg / mL of HCV virus genetically engineered antigen, and use subcutaneous multi-point injection for the initial immunization, 0.1 mL / point, a total of four points. 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 another 2 to 3 weeks, boo...

Embodiment 2

[0047] Embodiment 2 Hepatitis C indirect method ELISA diagnostic kit

[0048] Utilizing the bifunctional antibody obtained in Example 1 as a positive control to prepare a hepatitis C indirect enzyme immunoassay diagnostic kit, the composition is as follows:

[0049] 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 liquid

3ml×1 bottle

6ml×1 bottle

Store at 2-8°C

Chromogen B liquid

3ml×1 b...

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PUM

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Abstract

The invention relates to enzyme linked immunosorbent assay kits and in particular provides a bifunctional antibody serving as positive control in detection of hepatitis C virus. The bifunctional antibody is prepared from a mouse anti-HCV antibody and an anti-mouse IgG antibody by virtue of a chemical modification method. The bifunctional antibody prepared by utilizing an immunological method is taken as positive control of a hepatitis C indirect enzyme immunoassay diagnostic reagent, the source of the positive control material is solved, and the potential risk due to use of human serum is avoided.

Description

technical field [0001] The invention relates to an ELISA reagent kit, in particular to a bifunctional antibody used as a positive control for detecting hepatitis C virus 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)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18G01N33/577G01N33/576
Inventor 李银生王琳程珊赵林静张付平许锦秀朱杨阳
Owner XINXIANG MEDICAL UNIV
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