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Method for sceening viral nucleic acid of blood through isothermal amplification based on loop mediated technique

A virus nucleic acid and blood screening technology, applied in the field of clinical medicine, can solve problems such as missed detection

Active Publication Date: 2005-10-26
SHANGHAI BLOOD CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The existing blood screening method is mainly enzyme-linked immunoassay (ELISA), but due to the existence of the window period, it often leads to missed detection
It is known that the average window period of HCV antibody detection is 70 days, the average window period of HIV antibody detection is 22 days, and the average window period of HBsAg is 56 days. Missing detection during the window period of enzyme immunoassay is almost inevitable, which is the current further impact on blood safety. bottleneck of improvement

Method used

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  • Method for sceening viral nucleic acid of blood through isothermal amplification based on loop mediated technique
  • Method for sceening viral nucleic acid of blood through isothermal amplification based on loop mediated technique
  • Method for sceening viral nucleic acid of blood through isothermal amplification based on loop mediated technique

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0125] Example 1 Application example of the operating method of the HBV blood screening and identification system

[0126] Reagent

Packing volume Storage conditions

nucleic acid

Extraction reagent

Solution 1

300μl Store at 4°C

Solution 2

400μl Store at 4°C

Solution 3

500μl Store at 4°C

LAMP test

agent

Master Mix.

5ml Store at -20°C

Primer Mix.

3ml Store at -20°C

DNA polymerase

2ml Store at -20°C

Analyzing Software

LA-200 Program Ver0.18 software

[0127] The specific operation method is as follows:

[0128] (1) Nucleic acid extraction

[0129] Follow the steps below for HBV nucleic acid extraction

[0130] a. Add 300μl of solution to a microcentrifuge tube of 1 to 1.5ml

[0131] b. Add 100μl serum or plasma to the centrifuge tube and vortex for 5 seconds

[0132] c. Incubate at room temperature (15-25°C) for 10 minutes, boil for 10 minutes, and cool to room ...

Embodiment 2

[0150] Example 2 HBV, HCV and HIV triple blood screening identification system operating method

[0151] Reagent

Packing volume Storage conditions

nucleic acid

Extraction reagent

Solution 1

300μl Store at 4°C

Solution 2

400μl Store at 4°C

Solution 3

500μl Store at 4°C

LAMP test

agent

Master Mix.

5ml Store at -20°C

Primer Mix.

3ml Store at -20°C

Enzyme Mix

2ml Store at -20°C

Analyzing Software

LA-200 Program Ver0.18 software

[0152] The specific operation method is as follows:

[0153] (1) Nucleic acid extraction

[0154] Follow the steps below for HBV, HCV and HIV nucleic acid extraction

[0155] Add 300 μl solution to 1.5 ml microcentrifuge tube

[0156] Add 100μl serum or plasma to the centrifuge tube and vortex for 5 seconds

[0157] Incubate at room temperature (15-25°C) for 10 minutes

[0158] Add 400μl solution 2 to the centrifuge tube, and vo...

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PUM

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Abstract

This invention belongs to clinic domain, discloses a detecting method for screening viral nucleic acid of blood through isothermal amplification based on loop mediated technique, which is used to detecting nucleic acid virus selection of blood. We detect or select the nucleic acid consisting of HBV, HCV and HV by technology of molecular domain, this technology process is that: using special primer, using LMAP tech platform to amplify target gene under assistant of a series control system and inner comparison. This invent has the virtue of simple and convenient, economical and fast, delicacy and special, so that it has a expansive application.

Description

technical field [0001] The invention belongs to the field of clinical medicine, and relates to a blood screening method, more specifically, the invention relates to a blood virus nucleic acid screening method developed based on the principle of loop-mediated isothermal amplification technique (LAMP). Background technique [0002] The existing blood screening method is mainly enzyme-linked immunoassay (ELISA), but due to the existence of the window period, it often leads to missed detection. It is known that the average window period of HCV antibody detection is 70 days, the average window period of HIV antibody detection is 22 days, and the average window period of HBsAg is 56 days. Missing detection during the window period of enzyme immunoassay is almost inevitable, which is a further factor affecting blood safety. Improve the bottleneck. The results of the recently completed blood safety evaluation in Shanghai show that the blood safety in Shanghai is close to the level ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 王迅易进华郑岚伍晓菲裴爱琳
Owner SHANGHAI BLOOD CENT
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