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Probe for detecting circulating tumor DNA (Deoxyribonucleic Acid) of human breast cancer and application of probe

A technology for human breast cancer and breast cancer, which is applied in recombinant DNA technology, DNA/RNA fragments, and the determination/examination of microorganisms. Easy to collect, highly specific effects

Inactive Publication Date: 2016-09-21
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Breast ultrasonography has poor objectivity and needs to rely on the experience of the sonographer, which is highly subjective. Therefore, foreign guidelines do not recommend it as the main screening method.

Method used

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  • Probe for detecting circulating tumor DNA (Deoxyribonucleic Acid) of human breast cancer and application of probe
  • Probe for detecting circulating tumor DNA (Deoxyribonucleic Acid) of human breast cancer and application of probe
  • Probe for detecting circulating tumor DNA (Deoxyribonucleic Acid) of human breast cancer and application of probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Design of a specific selector for detection of human breast cancer and preparation of probes

[0042] 1. Design of a specific selector for human breast cancer detection

[0043] The human breast cancer-specific selector of the present invention includes all exon data and breast cancer mutation driver gene of 496 breast cancer patients in the TCGA (THE CANCER GENOME ATLAS) database, the length is 0.365Mb, and the specific length of each part is determined according to breast cancer The subtype and gene distribution are shown in Table 2 below, designed and verified by the School of Bioinformatics Science and Technology, Harbin Medical University.

[0044] Table 2

[0045] Subtype Selector(Mb) Basal-like 0.044069 Her2 0.040817 Luminal A 0.216391 Luminal B 0.091488 BRCA1 0.008220 BRCA2 0.011386 Total (after deduplication) 0.365177

[0046] Specifically, all the exome data of the 496 cases of breast cancer pa...

Embodiment 2

[0058] Example 2 A capture sequencing system for diagnosing human breast cancer

[0059] The capture sequencing platform of the present invention includes the probe prepared in Example 1, a hybridization system, a sequencing device (Roche Nimblegen) and a circulating free DNA extraction kit (QIAamp Circulating Nucleic Acid Kit).

Embodiment 3

[0060] Example 3 Application of the gene chip of the present invention in the diagnosis of human breast cancer

[0061] The capture sequencing platform used in this invention is Roche Nimblegen; the circulating free DNA extraction kit is QIAamp Circulating Nucleic Acid Kit; the sequencing analysis is done by Shanghai Ouyi.

[0062] The biological materials used in the present invention are all from the Department of Breast Surgery, Tumor Hospital Affiliated to Harbin Medical University, and all samples are provided with informed consent and ethics certificates signed by the patients themselves.

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Abstract

The invention discloses a probe for detecting a circulating tumor DNA (Deoxyribonucleic Acid) of a human breast cancer and application of the probe, belonging to the field of diagnosis and prevention of diseases. The invention designs a breast cancer selector which consists of DNA segments in mutation regions of related genes of the breast cancer. The circulating tumor DNA (ctDNA) of a breast cancer patient is subjected to liquid-phase hybridization capture by using the probe designed by the selector; after high-throughput sequencing, specific cancer gene variation of a patient is found, and a tumor load is monitored. Compared with a conventional detection means, the probe and the application thereof disclosed by the invention have the characteristics of no invasiveness, high throughout, good sensitivity and specificity, and the like; the probe and the application thereof can be used for better screening and early-stage diagnosis of the human breast cancer and are applied to individualized treatment of cancer.

Description

technical field [0001] The invention relates to a method for diagnosing diseases, in particular to a method for detecting human breast cancer circulating tumor DNA, which belongs to the field of disease diagnosis and prevention. Background technique [0002] Breast cancer is the malignant tumor with the highest incidence rate among women in the world. About 1.7 million women suffer from breast cancer and 522,000 die from breast cancer every year in the world. The incidence of breast cancer in the world varies with regional environment, economic differences and living habits. Generally speaking, developed countries such as Western Europe and North America have a high incidence rate, while most parts of Asia, Latin America, and Africa are low-incidence areas. From a global perspective, the annual incidence of breast cancer in the world increases by 0.2% to 8%. Breast cancer is the first cause of death in women aged 40 to 55. Since the late 1970s, the incidence of breast canc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869C12Q1/6886C12Q2600/156C12Q2565/519C12Q2535/122C12Q2525/191
Inventor 庞达张显玉肖云魏巍
Owner HARBIN MEDICAL UNIVERSITY
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