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A quantitative method for hbv-dna, a primer and probe for detecting hbv-dna, and a detecting kit comprising the same

A kit and probe technology, applied in the field of hepatitis B virus quantification, to achieve the effects of solving technical and economic problems, enhancing sensitivity, and improving signal strength

Inactive Publication Date: 2009-04-01
凯奇拜基因有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] [8] In particular, some Korean pharmaceutical companies (such as Dong-A Pharmaceutical, Green-Cross, Shin-Jin Medics, etc.) can produce antigen-antibody reaction-screening reagents that can only confirm whether a patient is affected by HBV, but there is no pharmaceutical The company develops and produces reagents for quantitative detection of HBV-DNA according to the molecular biology method of quantitative detection of HBV-DNA

Method used

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  • A quantitative method for hbv-dna, a primer and probe for detecting hbv-dna, and a detecting kit comprising the same
  • A quantitative method for hbv-dna, a primer and probe for detecting hbv-dna, and a detecting kit comprising the same
  • A quantitative method for hbv-dna, a primer and probe for detecting hbv-dna, and a detecting kit comprising the same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] [53] Example 1. Use of Enzyme Immunoassay Analyzer

[0059] [54] Composition of reagents

[0060] [55] 1. HBV-DNA extraction process

[0061] [56] 1) Dissolving reagent (02.N NaOH)

[0062] [57] 2) Neutralization reagent (0.2N HCl)

[0063] [58]

[0064] [59] 2. Amplification and hybridization reaction

[0065] [60] * Amplification of HBV-DNA

[0066] [61] 1) PCR reagents with freeze-dehydration (containing 2.5U of polymerase, 250 μM of four types of nucleic acid, 10 mM of Tris-HCl, 40 mM of KCl, MgCl 2 and stabilized colorant, and stored in a dehydrated state)

[0067] [62] 2) PCR buffer solution (purified water containing 100 μM primers and 1 pM d-UTP)

[0068] [63] 3) Low concentration positive control

[0069] [64] 4) Medium concentration positive control

[0070] [65] 5) Draw the standard sample of the standard curve

[0071] [66] Concentration of standard solution: 1.0.5pg / ml

[0072] [67] 2.100pg / ml

[0073] [68] 3.1000pg / ml

[0074] [69] 4.2000p...

Embodiment 2

[0112] [107] Example 2. Use of Chemiluminescence Meter

[0113] [108] Composition of reagents

[0114] [109] 1. HBV-DNA extraction process

[0115] [110] 1) Dissolution test solution (0.2N NaOH)

[0116] [111] 2) Neutralize test solution (0.2N HCl)

[0117] [112] 2. Amplification and hybridization reaction

[0118] [113] * Amplification of HBV-DNA

[0119] [114] 1) Freeze-dehydrated PCR reaction reagents (comprising 2.5U of polymerase, 250 μM of four types of nucleic acid, 10 mM of Tris-HCl, 40 mM of KCl, MgCl 2 and stabilized colorant, and stored in a dehydrated state)

[0120] [115] 2) PCR buffer solution (purified water containing 100 μM primers and 1 pM d-UTP)

[0121] [116] 3) Low concentration positive control

[0122] [117] 4) Medium concentration positive control

[0123] [118] 5) Draw the standard sample of standard curve

[0124] [119] Concentration of standard solution: 1.0.5pg / ml

[0125] [120] 2.100pg / ml

[0126] [121] 3.2000pg / ml

[0127] [122] 4....

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Abstract

The present invention relates to a method for quantifying hepatitis B virus DNA (HBA-DNA) by means of the PCR hybridization process, and to primers and probes used for such method. The present invention also relates to a kit for detecting HBV-DNA comprising said primers and probes. Specifically, the present invention is characterized in that said HBV is either an adr sub-type or an adw sub-type.

Description

technical field [0001] [1] The present invention relates to a method for quantifying hepatitis B virus (HBV) through a PCR hybridization process, and also relates to primers and probes used in the quantification method. The invention also relates to an HBV-DNA detection kit comprising the primer and probe. Background technique [0002] [2] Hepatitis B virus (HBV) is a virus from the family Hepadnaviridae that specifically acts on the human body. It is understood that about 500 million people in the world are infected with HBV. The incubation period of HBV is 60 to 110 days. After varying levels of clinical cycles, 90-95% of patients infected with HBV fully recover, but for patients who do not recover from infection, HBV genomic DNA is inserted and integrated into the patient's stem cell genomic DNA until it develops into Chronic active hepatitis, liver cirrhosis, liver cancer, etc. Like other diseases, HBV infection can cause chronic viral infection disease lymphadenoma ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12N15/11C12Q1/6851C12Q1/6883C12Q1/706
Inventor 崔贤一朴亨根金泰宇李惠泳赵孝卿
Owner 凯奇拜基因有限公司
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