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Screening of preinvasive carcinoma, carcinoma in situ and carcinoma focus by using DNA of circulating epstein-barr virus

A technology for precancer and cancer, which is applied in the field of screening of precancer, carcinoma in situ and cancer lesions by using circulating ibovirus DNA, which can solve the problems such as hindering the immune surveillance of tumor antigens.

Inactive Publication Date: 2006-06-21
杨华显
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these T cells were originally found to protect the host against the development of autoimmunity, they also prevent effective immune surveillance of the host against tumor antigens

Method used

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  • Screening of preinvasive carcinoma, carcinoma in situ and carcinoma focus by using DNA of circulating epstein-barr virus
  • Screening of preinvasive carcinoma, carcinoma in situ and carcinoma focus by using DNA of circulating epstein-barr virus
  • Screening of preinvasive carcinoma, carcinoma in situ and carcinoma focus by using DNA of circulating epstein-barr virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment I

[0047] Materials and methods

[0048] Blood samples from 132 high-risk cancer patients (Group A) and 3987 approximately normal individuals (Group B) have been sent to our laboratory in Hong Kong for the determination of circulating EBV DNA.

[0049] DNA Extraction from Plasma Samples Peripheral blood (5 ml) was collected from each subject and injected into EDTA tubes for separation of plasma. Blood samples were centrifuged at 1600 x g, and the plasma was carefully separated from the EDTA-containing tubes and transferred into blank polypropylene tubes. This sample was stored at -20°C until further manipulation. DNA was extracted from plasma samples using the QIAamp Blood Kit (Qiagen, Hilden, Germany) using the blood and body fluids procedure according to the manufacturer's recommendations (2). DNA extraction was performed using plasma samples (130-800 μl / column). Record the exact amount used to calculate the target DNA concentration. The volume of fina...

Embodiment II

[0066] Materials and methods

[0067] The percentage of CD4+CD25+ T cells in the peripheral blood of 20 patients with positive EBV DNA titers (group C) was compared with the EBV DNA-negative control group.

[0068] Peripheral blood from these 20 patients was obtained at the time of blood collection, injected into a Vacutainer tube containing heparin, diluted 2:1 with 1×Dulbecco's phosphate buffer (Mediatech) without calcium and magnesium, and then incubated at room temperature on poly Glycosomes (ficoll) were separated by centrifugation at 1000×g for 20 minutes on a density gradient. Peripheral blood mononuclear cells were collected, washed and cryopreserved in RPMI1640 containing 20% ​​fetal bovine serum and 10% DMSO for future use.

[0069] Cell types were determined using a four-color hemocytometer and flow cytometric analysis using activity markers of anti-CD3-PerCP, anti-CD4-APC or anti-CD8-FIT, anti-CD25-PE. Briefly, cells were incubated for 30 min...

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Abstract

The invention discloses a cancer shifting and finding method of different classifications and phases, whatever the cancer is relative to the Ibo-virus, which analyzes individual plasma or Ibo-virus DNA level in the serum once or several times continuously.

Description

technical field [0001] The present invention relates to the discovery that Epstein-barr virus (EBV) can be found in the cell-free fluid of a patient's blood, and when the virus is found, the patient may already have precancer, carcinoma in situ, or other Any form of, but not limited to, Ebovirus-associated cancer. The theory on which the present invention is based is that we found that circulating CD25+CD4+ regulates T(T reg ) Lymphocytes rise. We also found positive titers of circulating EBV DNA in the serum or plasma of patients with certain cancers of unknown relationship to Ibovirus. Combining the above observations, the findings demonstrate that EBV reactivation can lead to suppression of cytotoxic T cells, leading to a loss of cancer immune surveillance. The present invention proposes a new universal screening method, which uses circulating EBV DNA in serum or plasma to screen precancer, carcinoma in situ and cancer lesions. technical background [0002] It is well...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50C12Q1/68C12Q1/70G01N21/76
Inventor 杨华显
Owner 杨华显
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