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Hormone receptor-positive breast cancer recurrence monitoring gene mutation library construction method

A technology of hormone receptors and construction methods, applied in biochemical equipment and methods, microbial measurement/inspection, DNA preparation, etc., can solve problems such as interference, treatment decision-making troubles, and detection method distortion

Active Publication Date: 2017-07-11
XIAMEN SPACEGEN BIOTECH CO LTD
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Problems solved by technology

However, this kind of treatment decision is only based on the molecular classification determined by the initial treatment and the subjective judgment of whether the current clinical manifestation is "inert", and the molecular biological characteristics of the primary lesion of breast cancer and the metastasis have been proved to be significantly different; At the same time, once endocrine therapy is applied, it takes about 2 months on average for it to take effect, during which it is often disturbed by the phenomenon of "flickering", which leads to the distortion of routine detection methods; in addition, this type of advanced breast cancer often has multiple skin and bone metastases, and the above-mentioned lesions Difficult to objectively evaluate with existing imaging methods
These factors often cause great confusion to the treatment decision of this type of patients and limit the maximum benefit of endocrine therapy

Method used

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  • Hormone receptor-positive breast cancer recurrence monitoring gene mutation library construction method
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  • Hormone receptor-positive breast cancer recurrence monitoring gene mutation library construction method

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Embodiment Construction

[0144] The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

[0145] Construction method of gene mutation library for hormone receptor positive breast cancer recurrence monitoring, covering human genes KRAS, JAK3, AKT1, CREBBP, PTEN, RB1, TP53, CTNNB1, TSC2, TSC1, ERBB2, PIK3CA, SF3B1, JAK2, CDH1, SMAD4, A total of 1357 somatic mutations in GATA3, MAP2K4, MAP3K1 and ESR1.

[0146] Specifically include the following steps:

[0147](1) Design basic amplification primers for target genes KRAS, JAK3, AKT1, CREBBP, PTEN, RB1, TP53, CTNNB1, TSC2, TSC1, ERBB2, PIK3CA, SF3B1, JAK2, CDH1, SMAD4, GATA3, MAP2K4, MAP3K1 and ESR1 The 5' end of the forward primer and the reverse primer of the basic amplification primer set is provided with an additional 2-5 Ts, and the first T of the 2-5 Ts near the 3' end has a PNA modification, At the same time, the Tm value difference of the basic amplification primer se...

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Abstract

The invention discloses a hormone receptor-positive breast cancer recurrence monitoring gene mutation library construction method, which is characterized that the library covers the total 1357 somatic cell mutations on human genes such as KRAS, JAK3, AKT1, CREBBP, PTEN, RB1, TP53, CTNNB1, TSC2, TSC1, ERBB2, PIK3CA, SF3B1, JAK2, CDH1, SMAD4, GATA3, MAP2K4, MAP3K1 and ESR1. According to the present invention, a plurality of the target sequences are subjected to single tube amplification with the construction method to rapidly complete the library construction, wherein the whole library construction process takes only 2-3 h and the manual time only needs 45 min, such that the difficulty that the multi-gene and multi-target detection of somatic cells on the basis of the small amount of the peripheral blood sample is required in the clinical breast cancer recurrence monitoring is required can be effectively solved, and the lost is low.

Description

technical field [0001] The invention relates to a method for constructing a mutation library of a hormone receptor-positive breast cancer recurrence monitoring gene for high-throughput sequencing detection. Background technique [0002] Breast cancer will become the group with the largest number of cancer survivors in the future. According to the early molecular classification, there are two recurrence peak periods, which are 1-3 years and 5-10 years after surgery, during which repeated imaging examinations are required. Especially for hormone receptor-positive patients, endocrine therapy has been suggested to continue for 10 years, but there is no efficacy predictor that can accurately identify the beneficiary group, and there are many complications. Therefore, it is necessary to develop a non-invasive efficacy prediction and monitoring system for breast cancer It will reduce the radiological dose of patients and reduce unnecessary side effects of treatment. Traditional tu...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/10
CPCC12N15/1068C12Q1/6886C12Q2600/156
Inventor 辇伟奇张海伟李丽仙余慧青唐万燕张娜何永鹏
Owner XIAMEN SPACEGEN BIOTECH CO LTD
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