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Stable preparation method for antigen slices used in detection of FAMA (Fluorescent Antibody to Membrane Antigen)

An antigen and stable technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of large errors, undetected, short shelf life of antigen sheets, etc.

Active Publication Date: 2018-06-19
CHANGCHUN KEYGEN BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the traditional FAMA method, each laboratory uses the Oka strain of varicella-zoster virus to infect diploid cells to prepare antigen sheets, and the antigen sheets prepared by the traditional method have a short shelf life. Fast inactivation cannot bind to specific antibodies
In a patent for a preparation method of antigen sheets for varicella virus antibody detection, the antigen sheet is prepared by infecting VERO cells with the VZV84-7 strain, which can be stored in a low-temperature or ultra-low temperature refrigerator for up to one year. However, general laboratories cannot meet this condition and are limited. application of the method
The antigen sheet prepared by this method will quickly inactivate the virus cell sheet when stored at 2-8°C, and it is very difficult to use the same batch of antigen sheets in large quantities in the experiment. Using antigen sheets prepared many times to detect the same batch of large batches of VZV serum samples has a large error
Up to now, there are no literature reports on how to prolong the preservation of antigen sheets and the new method of preparing antigen sheets.

Method used

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  • Stable preparation method for antigen slices used in detection of FAMA (Fluorescent Antibody to Membrane Antigen)
  • Stable preparation method for antigen slices used in detection of FAMA (Fluorescent Antibody to Membrane Antigen)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Add 30 μl of protective solution containing 2% lactose and 3% trehalose (the solution for dissolving sugar is 20 mmol / l PBS, pH value 7.4) dropwise to each well of the surface of the VZV antigen sheet cells prepared in the above manner, wait for drying, and pump Vacuum seal the slides and store at 2-8°C for later use.

Embodiment 2

[0024] On the surface of the VZV antigen sheet cells prepared in the above manner, 30 μl of protective solution containing 2% lactose, 3% trehalose and 2% dextran were added dropwise to each hole (the solution for dissolving sugar was 50 mmol / l PBS, pH value 7.0), after drying, vacuum seal, and store at 2-8°C for later use.

Embodiment 3

[0026] Add 20 μl of protective solution containing 1% lactose and 4% trehalose (the solution for dissolving sugar is 20 mmol / l PBS, pH value 7.4) to each well of the surface of the VZV antigen sheet cells prepared in the above-mentioned way, and wait for drying and vacuumize Seal the slides and store at 2-8°C for later use.

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Abstract

The invention provides an FAMA (Fluorescent Antibody to Membrane Antigen) antigen slice preparation and preservation method for detecting varicella zoster virus (VZV) antibodies. The method comprisesthe following steps: infecting 2BS cells with varicella zoster virus strains, incubating in a 5% of CO2 incubator at a temperature of 35 DEG C to reach a virus replication peak, performing trypsin digestion and cell counting, selecting an appropriate cell quantity to prepare virus cell suspension, dropping the suspension onto a 12-well glass slide, incubating in a 5% of CO2 incubator at a temperature of 37 DEG C, performing cold acetone fixation, cleaning, drying, dropping a protective fluid on the cell surface to seal, and vacuumizing. The antigen slices can be preserved in a refrigerator of2-8 DEG C for one year or longer.

Description

technical field [0001] The invention relates to a stable preparation method for antigen sheet detection by FAMA method and the prepared antigen sheet. It specifically relates to a cell that is infected with varicella zoster virus until the cell is damaged, digested to make a cell suspension, drop the tablet, and after drying, add a protective solution dropwise, and after drying, vacuum seal the film, which can be stored at 2-8°C A preparation method for long-term preservation. Background technique [0002] The FAMA method is currently internationally recognized as the gold standard for the detection of varicella-zoster virus antibodies, and has the characteristics of high sensitivity and strong specificity. However, the preparation and storage of the antigen sheet is the decisive part of this experiment. [0003] In the traditional FAMA method, each laboratory uses the Oka strain of varicella-zoster virus to infect diploid cells to prepare antigen sheets, and the antigen s...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56994
Inventor 朱晓文
Owner CHANGCHUN KEYGEN BIOLOGICAL PROD
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