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A kit for auxiliary diagnosis of early early liver cancer and a detection method thereof

A technology for early stage liver cancer and auxiliary diagnosis, which is used in the determination/examination of microorganisms, biochemical equipment and methods, instruments, etc.

Inactive Publication Date: 2019-01-11
杭州翱锐生物科技有限公司
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Problems solved by technology

Liquid biopsy mainly relies on sensitive technology to detect tumor-specific DNA (circulating tumor DNA, circulating tumor DNA, ctDNA) in the free DNA (ccfDNAs) in the blood; image 3 As shown, since the amount of tumor DNA released into the blood is closely related to the size of the tumor and the clinical stage of the tumor, while early tumors release very little DNA into the blood, it is a great challenge to diagnose early tumors with liquid biopsy

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  • A kit for auxiliary diagnosis of early early liver cancer and a detection method thereof
  • A kit for auxiliary diagnosis of early early liver cancer and a detection method thereof
  • A kit for auxiliary diagnosis of early early liver cancer and a detection method thereof

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Embodiment Construction

[0067] The present invention will be specifically introduced below in conjunction with the accompanying drawings and specific embodiments.

[0068] A kit for auxiliary diagnosis of early liver cancer, including the following reagents: ccfDNA end treatment system, circular linker reaction system, 0.06-0.15U / ul USER enzyme, PCR amplification system, magnetic beads. ccfDNA: circulating cell-free DNA.

[0069] The ccfDNA end treatment system consists of: ccfDNA, 0.003-0.007U / ul T4 DNA polymerase, 0.1-0.3U / ul T4 polynucleotide kinase, 0.0015-0.0035U / ul TaqDNA polymerase, 1x T4 DNA ligase Buffer, 0.2-0.6 mM dNTP, 0.1-0.3 mM ATP, 2.5% binder.

[0070] The circular linker reaction system consists of: 0.1-0.3U / ul T4 ligase, 1xT4 ligase buffer, and 0.7-1.2uM circular linker, the circular linker sequence: 5'-P-GAT CGG AAG AGC ACA CGT CTG AAC TCC AGTC dU A CAC TCT TTC CCT ACA CGA CGC TCT TCC GAT CT-3'.

[0071] The PCR amplification system consists of 15-25 parts of DNA fragments, 3-8 ...

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Abstract

The invention discloses a kit for auxiliary diagnosis of early liver cancer and a detection method thereof, which comprises the following reagents: a ccfDNA terminal treatment system, a ring linker reaction system, 0.06-0.15 U / ul USER enzyme, PCR amplification system, magnetic beads; The whole ccfDNA genome is sequenced by sequencing library. After genome-wide data processing, statistics and machine learning model were established to detect the abnormal copy number of ccfDNA in patients so as to achieve the early diagnosis of liver cancer. Such a detection method maximizes the accuracy of fluid biopsy for the diagnosis of early liver cancer, especially for the detection of primary liver cancer.

Description

technical field [0001] The invention relates to the field of auxiliary diagnosis of liver cancer, in particular to a kit for auxiliary diagnosis of early liver cancer and a detection method thereof. Background technique [0002] Liver cancer is a highly malignant tumor. Once it metastasizes, there is basically no effective treatment plan, and the survival rate is very low. If liver cancer is diagnosed at an early stage, surgery or liver transplantation can still be performed, and the five-year survival rate can reach 70%. If it has been transferred, the five-year survival rate is only about 5%. So early diagnosis is crucial to the patient's survival. [0003] Routine diagnosis of liver cancer is mainly through alpha-fetoprotein (AFP) screening. For high-risk groups, ultrasound, MRI and other imaging methods are used for further examination. When suspicious lesions are found, the diagnostic method is liver biopsy. However, ultrasound has limited sensitivity and is not defi...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6869G16B40/10G16B30/00
CPCC12Q1/6869C12Q1/6886C12Q2600/156C12Q2531/113C12Q2535/122C12Q2537/16C12Q2537/165
Inventor 杨楚虎张琼
Owner 杭州翱锐生物科技有限公司
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