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Blood plasma cfDNA detection technology-based noninvasive pancreas cancer polygene detection method and kit

A detection technology and detection method technology, which are applied in the field of non-invasive pancreatic cancer multi-gene detection methods and kits

Inactive Publication Date: 2016-11-02
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, there are hundreds of copies of mitochondrial DNA in each human cell, and the detection limit of traditional Sanger sequencing technology is 10-15%, so many low-abundance mitochondrial heterogeneities cannot be detected by traditional techniques

Method used

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  • Blood plasma cfDNA detection technology-based noninvasive pancreas cancer polygene detection method and kit
  • Blood plasma cfDNA detection technology-based noninvasive pancreas cancer polygene detection method and kit
  • Blood plasma cfDNA detection technology-based noninvasive pancreas cancer polygene detection method and kit

Examples

Experimental program
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Effect test

Embodiment 1

[0058] The whole blood (provided by the First Affiliated Hospital of Zhejiang University) from 3 patients with pancreatic cancer from No. 1-3 was subjected to a pancreatic cancer plasma cfDNA polygene test and a tumor tissue DNA polygene test to compare their consistency and judge Whether the mutations detected in plasma cfDNA can be used as individualized monitoring markers for patients.

[0059] 1. Take material, separate

[0060] 1.1 5ml of whole blood from 3 pancreatic cancer patients from 1-3, add the whole blood to the EDTA tube containing anticoagulant. Transfer the EDTA tube to a 4°C refrigerated centrifuge within 2 hours and centrifuge at 1900g (3000rpm) for 10 minutes. The lower layer contains blood cells and the upper layer contains plasma. They are transferred to EP tubes and stored at -80°C for storage.

[0061] 1.2 Take 30 mg of tumor surgery tissue and put it directly into the EP tube and immediately freeze it at -80°C or liquid nitrogen.

[0062] 2 DNA purification an...

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Abstract

The invention belongs to the field of medicines and biology, and relates to a blood plasma cfDNA detection technology-based noninvasive pancreas cancer polygene detection method and kit. The blood plasma cfDNA detection technology-based noninvasive pancreas cancer polygene detection method comprises the following steps: collecting blood, separating blood plasma, and purifying cfDNA; designing the panel of a pancreas cancer characteristic hotspot mutation region; and carrying out PCR primer amplification, constructing a library, and carrying out high-flux next generation sequencing and data analysis. The dynamic change of the mutation type and the mutation frequency of pancreas cancer characteristics in the blood plasma cfDNA is detected according to the treatment process and the pathogenesis development of every individual, so information is provided for pathogenesis monitoring and resistance drugs of the pancreas cancer.

Description

Technical field [0001] The invention belongs to the field of medical biology and relates to a non-invasive pancreatic cancer multi-gene detection method and kit based on plasma cfDNA detection technology. Background technique [0002] Pancreatic cancer is one of the common malignant tumors of the digestive system. It usually refers to pancreatic tubular adenocarcinoma. Its characteristics such as insidious onset, difficult early diagnosis, rapid progress, and easy recurrence and metastasis have led to poor clinical treatment effects, and it is currently the worst-prognosis malignant tumor One, the median survival period is about 6 months, and the overall 5-year survival rate is 3 to 5%. It is a serious threat to human health and is called the "king of cancer." [0003] Studies have shown that human cancer is caused by the accumulation of oncogenes and tumor suppressor gene mutations. The vast majority of gene mutations are non-functional or non-pathogenic mutations, called "Passeng...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/106C12Q2600/118C12Q2600/156
Inventor 王伟林赵忻艺周东锴胡振华叶松孔阳
Owner ZHEJIANG UNIV
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