Breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination and kit thereof

A detection kit, PIK3CA542-MT3 technology, applied in the field of molecular biology, can solve the problems of being easily affected by various factors, low throughput and high cost

Active Publication Date: 2018-12-18
SHENZHEN HUADA GENE INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, gene sequencing is internationally recognized as the most standard and accurate, but the cost is high and the throughput is small, which is more conducive to the analysis of complex and highly polymorphic genes
The advantage of the gene chip lies in its automation and high throughput.

Method used

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  • Breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination and kit thereof
  • Breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination and kit thereof
  • Breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination and kit thereof

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Experimental program
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Embodiment Construction

[0044] The present invention will be further described in detail below through specific embodiments in conjunction with the accompanying drawings.

[0045] The present invention is mainly based on a droplet digital PCR platform, and designs primers and probes for three mutation sites of the PIK3CA gene, and uses cell line DNA and positive standard products as samples for testing and optimization, thereby determining the reaction system. The specific detection sites are shown in Table 1 below:

[0046] Table 1

[0047]

[0048] The detection primer sequences of PIK3CA mutation sites are shown in Table 2:

[0049] Table 2

[0050]

[0051]

[0052] The detection probe sequence of the PIK3CA mutation site is shown in Table 3, in which the FAM and VIC fluorophores are located at the 5' end.

[0053] table 3

[0054]

[0055] The technical scheme of the present invention will be described in detail below through examples.

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Abstract

The invention discloses a breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination and a kit thereof. The detection probe and primer sequence combination comprises a primer sequence as shown in SEQ ID NO: 1-4, and a probe sequence as shown in SEQ ID NO: 5-10. The probe with the sequence is modified with a fluorophore. Based on a micro-drop digital PCR platform, primersand probes are designed for three mutation sites of the PIK3CA gene; and through experimental verification, a breast cancer PIK3CA hotspot mutation detection probe and primer sequence combination isdetermined and the combination can effectively detect the three mutation sites of the PIK3CA gene.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a primer sequence combination and a kit for breast cancer PIK3CA hotspot mutation detection probes. Background technique [0002] Breast cancer is a common malignant tumor that seriously threatens women's health. Its incidence is increasing year by year, and it has become the number one malignant tumor in women. With the continuous deepening of research on molecular biological changes in the occurrence and development of breast cancer, targeted therapy using tumor cell signal transduction pathways as the treatment site has become another effective treatment for breast cancer after chemotherapy and endocrine therapy. treatment approach. [0003] The phosphatidylinositol-3-kinase catalytic unit (phosphatidylinositol-3-kinase catalyticalpha, PIK3CA) was detected by Volinia in 1994 using in situ hybridization. It is located at 3q26.3, 34kb long, and contains 21 exons sub,...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/686C12Q1/6886C12Q2600/156C12Q2563/159C12Q2563/107
Inventor 李南南易吉朱师达
Owner SHENZHEN HUADA GENE INST
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