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Method for fast inspecting HBV, HCV, HIV, TP and RV simultaneouslly

An HIV-1, rapid technology, applied in the field of detection and diagnosis, which can solve the problems of patients infected with major pathogenic microorganisms, infection, reduction, etc.

Inactive Publication Date: 2007-04-04
鲍玉洲 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the development of urban construction in recent years, more and more people suffer from corneal trauma, but the corneal donor materials that can be used for corneal transplantation are decreasing day by day. The possibility of pathogenic microorganisms, which puts patients at risk of infection with serious pathogenic microorganisms through iatrogenic routes

Method used

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  • Method for fast inspecting HBV, HCV, HIV, TP and RV simultaneouslly
  • Method for fast inspecting HBV, HCV, HIV, TP and RV simultaneouslly
  • Method for fast inspecting HBV, HCV, HIV, TP and RV simultaneouslly

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] Embodiment 1, corneal virus detection:

[0105] 1. Collection of materials

[0106] Quickly cut about 50 mg of peripheral corneal tissue material from the donor, shred it with scissors under sterile conditions, and grind it thoroughly in a grinder to lyse the tissue and release the cells.

[0107] 2. Extraction of genomic DNA (refer to the Qiagen kit operation steps for extraction)

[0108] 1. Add 20ul proteinase K to the bottom of a 1.5ml centrifuge tube;

[0109] 2. Add 200ul sample to the centrifuge tube and mix well;

[0110] 3. Add 200ul Buffer AL to the sample, and vortex for 15s to mix;

[0111] 4. Incubate at 56°C for 10 minutes;

[0112] 5. Slightly centrifuge to remove the liquid sticking to the tube cap;

[0113] 6. Add 200ul of absolute ethanol to the sample, vortex for 15s to mix, then centrifuge slightly to remove the liquid sticking to the tube cap;

[0114] 7. Carefully transfer the sample in step 6 to the filter column, without sticking to the col...

Embodiment 2

[0124] Embodiment 2, corneal virus detection

[0125] 1. Collection of materials

[0126] Quickly cut about 50 mg of peripheral corneal tissue material from the donor, shred it with scissors under sterile conditions, and grind it thoroughly in a grinder to lyse the tissue and release the cells.

[0127] 2. Extraction of genomic RNA (refer to Qiagen kit operation steps for extraction)

[0128] 1. Add 3.5ul β-mercaptoethanol to 350ulRLT;

[0129] 2. Take an appropriate amount of positive specimen (generally 100ul of serum; about 10mg of tissue, grind it in an RNase-free grinder in advance, and dilute it with 80ul PBS; feces, saliva, throat swabs, etc. need to be diluted with PBS) and add it to RLT, mix uniform;

[0130] 3. Centrifuge at 10000rpm for 5min, suck the supernatant into a new centrifuge tube;

[0131] 4. Add one volume (generally 350ul or 600ul) of 70% ethanol and mix:

[0132] 5. Take 700ul sample and put it on the column (2ml- column), centrifuge at 10000rpm fo...

Embodiment 3

[0145] Embodiment 3, corneal virus detection

[0146] 1. Collection of materials

[0147] Quickly cut about 50 mg of peripheral corneal tissue material from the donor, shred it with scissors under sterile conditions, and grind it thoroughly in a grinder to lyse the tissue and release the cells.

[0148] 2. Extraction of genomic RNA (refer to Qiagen kit operation steps for extraction)

[0149] 1. Add 3.5ul β-mercaptoethanol to 350ulRLT;

[0150] 2. Take an appropriate amount of positive specimen (generally 100ul of serum; about 10mg of tissue, grind it in an RNase-free grinder in advance, and dilute it with 80ul PBS; feces, saliva, throat swabs, etc. need to be diluted with PBS) and add it to RLT, mix uniform;

[0151] 3. Centrifuge at 10000rpm for 5min, suck the supernatant into a new centrifuge tube;

[0152] 4. Add one volume (generally 350ul or 600ul) of 70% ethanol and mix:

[0153] 5. Take 700ul sample and put it on the column (2ml- column), centrifuge at 10000rpm fo...

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PUM

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Abstract

A method for inspecting HBV, HCV, HIV, TP and RV simultaneously and rapidly is carried out by extracting virus inspection template, RNA virus reversed transcripting, PCR amplifying for five peccant microbe genes in the one same reactive system, inspecting by agarose gel electrophoresis method, positioning several peccant microbe gene sensitive target genes, designing multiple PCR nested or semi-nested primer and inspecting sensitively and rapidly. It can be used to inspect clinical human body or animal blood or tissue sample.

Description

technical field [0001] The invention belongs to the technical field of detection and diagnosis, and particularly relates to a rapid and sensitive detection of hepatitis B virus (Hepatitis B virus, HBV) and hepatitis C virus infection in humans or higher organisms simultaneously in a reaction system through polymerase chain reaction technology. (Hepatitis C virus, HCV), AIDS virus type I and type II (Human immunodeficiency virus type I and type II, HIV-1 and HIV-2), Treponema pallidum (Treponema pallidum, TP) and rabies virus (Rabies virus, RV) Methods. Background technique [0002] Hepatitis B, also known as serum hepatitis, is an infectious disease caused by the hepatitis B virus (HBV). It can be transmitted through blood, body fluids, and sexually, and has a chronic carrier state. The disease is widely prevalent in my country, and the infection rate of the population is high, and the infection rate in some areas is as high as 35%. The latest research data shows that the...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 鲍玉洲张艳敏
Owner 鲍玉洲
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