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Kit for detecting integrated viruses of genome in hybrid capture

A hybrid capture and genome technology, applied in the field of biomedicine, can solve the problems of inability to popularize clinical practice, produce non-specific products, and consume a lot of labor.

Inactive Publication Date: 2011-02-16
何以丰
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These restrictive conditions determine that this method is only suitable for some established cell lines and cannot be extended to real clinical practice.
[0008] 3) The genomic library method not only needs to obtain established tumor cell lines, but also consumes a lot of labor to carry out large-scale screening and sequencing identification of the obtained library clones
However, although PCR can guarantee the sensitivity of molecular detection, there is also the possibility of non-specific products during the amplification process.

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  • Kit for detecting integrated viruses of genome in hybrid capture
  • Kit for detecting integrated viruses of genome in hybrid capture
  • Kit for detecting integrated viruses of genome in hybrid capture

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Embodiment Construction

[0042] First assemble the integrated HBV detection kit, including 8 components: 1) "DNA single-strand molecular capture mixture"; 2) Positive quality control; 3) Negative quality control; 4) "Positive quality control" specificity PCR amplification system with primer pair; 5) PCR amplification system with specific primer pair for “negative quality control”; 6) PCR amplification system with HBV specific primer pair; 7) “positive” and “negative” Control product" specific Tagman molecular probe; 8) HBV specific Tagman molecular probe.

[0043] (1) from 5×10 6 DNA was extracted from HT-3, HEK293, SKOV-3, CAOV-3, and OMC cells (using QIAamp DNA Mini Kit from Diagen Company), and then mixed. The DNA mixture was digested with Hae III endonuclease for 2 hours at 37°C, followed by inactivation at 90°C for 20 minutes. The resulting DNA fragments were then coupled to 1 g of Sephacryl S-500 (product of GE) solid phase carrier through diazotization. This is: "DNA Single Strand Molecular ...

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Abstract

The invention belongs to the field of biomedicine, in particular to a kit for detecting integrated viruses of a genome in hybrid capture. The kit comprises the following main components: (1) a solid phase medium fixed with a capture mixture of human genome DNA (Deoxyribonucleic Acid) single-chain molecules, (2) positive and negative quality control materials for identifying the hybrid capture efficiency and specificity, and (3) a primer pair and a molecular probe for carrying out real-time PCR (Polymerase Chain Reaction) identification to the captured mixture of the target DNA molecules, obtained by capture. By utilizing all or partial components in the invention, a complete integrated genome virus detection kit or an integrated genome virus detection kit with the type specificity can be assembled. The obtained virus detection kit can be used for detecting and identifying the integrated state of the genome in a certain common integrated and oncogenic viruses, such as human hepatitis B viruses, human papilloma viruses, Epstein-Barr viruses and human herpes simplex viruses, in tissue or cytology specimens.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a kit for detecting genome-integrated viruses by hybridization capture. Background technique [0002] Integration of viral DNA into the genome of human cells is a late event of infection. Common virus integration phenomena can be divided into two categories, one is the integration of deoxyribonucleic acid (abbreviated DNA) virus, such as: hepatitis B virus (Hepatitis B virus, abbreviated HBV), Epstein-Barr virus (Epstein-Barr virus) virus, abbreviated as EBV), adeno-associated virus (Adenovirus-associated virus, abbreviated as AAV) and human papillomavirus (Humanpapillomavirus, abbreviated as HPV); the other type is the integration of ribonucleic acid (abbreviated as RNA) virus, such as: Hepatitis C virus (abbreviated HCV) and human immunodeficiency virus (Humanimmunodeficiency virus, abbreviated HIV). DNA virus integration is mainly achieved by the already expressed integrase or trans...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12R1/93C12Q1/70
Inventor 何以丰
Owner 何以丰
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