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T cell immune state detecting method for hepatitis B patient

A technology for cell immunity and state detection, applied in measurement devices, color/spectral property measurement, instruments, etc., can solve the problems of easy inactivation, inability to guarantee the consistency of experimental conditions, poor stability of recombinant proteins, etc.

Inactive Publication Date: 2005-01-26
SHENZHEN DAKEWE BIOENG
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Problems solved by technology

The experimental method established in this way has poor specificity and high experimental background
[0007] 3. The stability of the recombinant protein is poor, and it is easy to be inactivated. Generally, it needs to be stored below minus 20 degrees.
Even if it is freeze-dried, it needs to be re-dissolved and cannot be used repeatedly
Thus, the consistency of the experimental conditions cannot be guaranteed

Method used

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  • T cell immune state detecting method for hepatitis B patient

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Embodiment Construction

[0011] This method is a detection method based on ELISPOT, and it mainly has the following requirements for ELISPOT:

[0012] 1. Select a coated monoclonal antibody against human interferon-γ with high activity and specificity. After the antibody is coated, it should be able to store its activity at 4 degrees for more than one year.

[0013] 2. Select the second monoclonal antibody against human interferon-γ with high activity and specificity. The antibody should be aimed at different antigen sites from the monoclonal antibody used for coating, and there will be no steric hindrance effect in binding to the antigen. After the antibody is covalently linked to biotin, it has high activity and can be stored at 4 degrees for more than one year.

[0014] 3. A commercially available 96-well highly absorbent cell culture plate and avidin-horseradish peroxide complex were used. All other laboratory reagents must be of analytical grade.

[0015] 4. Hepatitis B virus core antigen pep...

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Abstract

This invention discloses a detecting method for immune status of T-cell of hepatitis B patients. It is to use artificial synthesized polypeptide to stimulate the peripheral blood lymphocyte cultivated out of hepatitis B patients; to detect the number of lymphocyte with Gamma interferon in simulated body by use of enzyme associated immune blob experimental method, so to determine immune status of T-cell of hepatitis B patients. In normal matched group, there is barely no non-specific blot, and the stability of biological activity of synthesized polypeptide between batches is greatly increased and errors between batches are greatly induced. Synthesized polypeptide can be kept well in the temperature of four degree for six months without apparent reduction of activity, which is very helpful for the use of reagent box.

Description

Technical field: [0001] The invention relates to a method for detecting T cell immune state of hepatitis B patients. Background technique: [0002] Hepatitis B virus (HBV) infection is a global problem. In my country alone, more than 100 million people are long-term carriers of hepatitis B virus. Long-term carriers of the virus, especially those with the "big three yang" (surface antigen positive, e antigen positive, core antibody positive); will eventually lead to liver cirrhosis and liver cancer. Therefore, the treatment of hepatitis B is an important and urgent work. Unfortunately, humans have not found an effective method to cure hepatitis B virus so far. Studies in recent years have found that the body's response to viruses is closely related to its own immune status. When the T cells in the patient's immune system are in an activated state to the hepatitis B virus, the body will inhibit the virus with the cooperation of drugs, and ultimately facilitate the removal o...

Claims

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Application Information

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IPC IPC(8): G01N21/27G01N33/543
Inventor 吴庆军
Owner SHENZHEN DAKEWE BIOENG
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