Method for real-time detecting HBV through PCR

A composition and base sequence technology, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems such as reduced detection sensitivity, false negatives, and reduced amplification efficiency

Inactive Publication Date: 2010-10-27
LG LIFE SCI LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, false negative results or reduced detection sensitivity may be obtained due to a decrease in amplification efficiency
Therefore, it is difficult to guarantee accurate quantification

Method used

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  • Method for real-time detecting HBV through PCR
  • Method for real-time detecting HBV through PCR
  • Method for real-time detecting HBV through PCR

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1. Construction of primers and probes

[0066] The HBV-specific primers and probes used in the present invention are primer sequences capable of amplifying only HBV viral genes, which were analyzed by using the DNAsis program from Hitachi Software GenBank ( www.ncbi.nlm.nih.gov ) (managed by the National Center for Biotechnology Information (NCBI) at the National Institutes of Health (NIH)), this DNA sequence was sequenced, and subsequently used BLAST ( www.ncbi.nlm.nih.gov / BLAST / ) to analyze this DNA sequence again to confirm.

[0067] After the base sequence was obtained, HS-Taq was used to determine the specific primers and probes of plant-derived genes that were not competitively inhibited by the HBV-specific primers and probes in the HBV detection composition.

Embodiment 2

[0068] Example 2. Synthesis of primers

[0069] Using, for example, the third edition of Molecular cloning (Molecular cloning 3 rd , Sambrook and Russell, Cold Spring Harbor Laboratory Press, New York, USA, 2001) in the "synthesis of oligonucleotides" described in section 10.42, the primers analyzed in Example 1 were synthesized by Metabion (Germany).

Embodiment 3

[0070] Example 3. Extraction of HBV RNA from clinical samples

[0071] Serum or EDTA-plasma was isolated from blood of suspected HBV-infected patients and extracted using silica membrane-based spin columns such as the QIAamp MineElute virus spin column (Qiagen).

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PUM

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Abstract

The invention relates to a method for detecting HBV genes, in particular to a primer pair complemented with the HBV gene, a HBV detecting compound containing the primer pair complemented with the HBV gene and a probe, a HBV detecting kit containing the compound and a method for detecting HBV genes.

Description

technical field [0001] The invention relates to the detection of hepatitis B virus gene. More specifically, the present invention relates to a pair of primers complementary to HBV genes, a composition for HBV detection comprising a pair of primers complementary to HBV genes and a probe, an HBV detection kit comprising said composition, and a composition for detecting HBV genes Methods. Background technique [0002] Hepatitis B virus (hereinafter referred to as HBV) is known to be a major cause of viral hepatitis, and it is estimated that 2 billion people are infected with HBV worldwide. Of these infected populations, approximately 350 million are chronic carriers, and most are at risk of progressively developing cirrhosis and hepatocellular carcinoma. The HBV envelope gene consists of the preS region (including preS1 and preS2) and the S region. After transcription of the entire envelope gene, three envelope proteins (L protein, M protein, and S protein) are synthesized b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/70C12Q1/68G01N21/64
CPCC12Q1/706
Inventor 李东焕姜镇锡朴荣石金东铉
Owner LG LIFE SCI LTD
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