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Sequence combination and probe for simultaneously detecting multiple gene mutation types of lung cancer

A gene sequence and type technology, applied in the field of sequence combinations and probes for the simultaneous detection of multiple gene mutation types of lung cancer, can solve the problems of lung cancer patients' biopsy sampling volume that cannot meet the detection needs, delaying patient treatment, and prolonging the detection cycle. Shorten the detection process, improve the sensitivity and accuracy, and the effect of high sensitivity

Inactive Publication Date: 2018-01-23
广州漫瑞生物信息技术有限公司
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  • Summary
  • Abstract
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AI Technical Summary

Problems solved by technology

According to the report in the "Expert Consensus on Epidermal Growth Factor Receptor Gene Mutation Detection in Patients with Non-Small Cell Lung Cancer in China (2016 Edition)", there are two types of EGFR gene detection: sensitive mutations and drug-resistant mutations. Sensitive mutations include: exon 19 Deletion mutation (19del), exon 21 point mutation (L858R), G719X, L861Q, and S768I; drug resistance mutations include: exon 20 insertion mutation (20ins) and exon 20 point mutation (T790M), a total of about 31 Multiple types of mutations need to be detected, which requires a large number of DNA samples, and the amount of biopsy samples of some lung cancer patients cannot meet the detection requirements
Patients can only undergo ALK fusion gene detection again after multiple negative EGFR gene tests, which will inevitably prolong the detection cycle, delay the disease, and delay the patient's treatment

Method used

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  • Sequence combination and probe for simultaneously detecting multiple gene mutation types of lung cancer
  • Sequence combination and probe for simultaneously detecting multiple gene mutation types of lung cancer
  • Sequence combination and probe for simultaneously detecting multiple gene mutation types of lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Based on the obtained information of 62 hotspot mutation genes related to lung cancer, the present invention constructs probes and detection kits for 234 highly mutated regions of 62 hot spot mutation genes related to lung cancer.

[0042] The design of the probe is based on the NCCN guidelines, expert consensus, relevant literature reports, and information from medical websites such as ClinVar and Cosmic to screen genes, mutation sites or related regions, determine the target region of the probe, and design a synthetic probe for the simultaneous detection of lung cancer The sequence combinations of various gene mutation types are shown in Table 1.

[0043] Table 1

[0044]

[0045]

[0046]

[0047]

[0048]

[0049] The length of the probe is preferably 22.5kb, and the probe with this length can detect as many mutations as possible and further improve the detection efficiency.

[0050]First, the extracted sample DNA is broken into 200-250bp DNA fragmen...

Embodiment 2

[0155] In this example, the commercial standard Multiplex I cfDNAReference Standard Set from Horizon Discovery was used as the sample to be tested. Catalog#: HD778 is a ctDNA standard product with a mutation frequency of 1%, and Catalog#: HD779 is a ctDNA standard product with a mutation frequency of 0.1%. The kit of the invention is used to capture the standard DNA and build a library. After bioinformatics data analysis Obtain the detection result of the standard product DNA, and compare it with the known site frequency to verify the detection sensitivity and specificity of the detection kit of the present invention. This example protocol includes three aspects: pre-hybridization library construction, hybridization capture, sequencing and data analysis.

[0156] (1), library construction before hybridization, see Example 1 for the experimental operation steps.

[0157] (2), hybrid capture, see example 1 for the experimental operation steps.

[0158] (3), data analysis

[0...

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Abstract

The invention discloses a sequence combination and a probe for simultaneously detecting multiple gene mutation types of lung cancer. The sequence combination, the probe, a gene chip and a kit, which are provided by the invention and are high in sensitivity and specificity, can be used for simultaneously detecting the mutation types such as point mutation, short fragment insertion deletion, copy number variation and fusion gene mutation of 62 related hot spot mutation genes. The method provided by the invention has the characteristics of being comprehensive, rapid, high in flux, cost performance and sensitivity, strong in specificity, and the like, and greatly improves the detection efficiency.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to sequence combinations and probes for simultaneously detecting multiple gene mutation types of lung cancer. Background technique [0002] Primary lung cancer (referred to as lung cancer) is one of the most common malignant tumors in my country. According to the World Health Organization (WHO) cancer Globocan database, in 2012, the number of new lung cancer patients in my country reached 652,800, ranking first among all malignant tumors, accounting for about 1 / 3 of the world, and more than 80% of the patients were non-small cell lung cancer (non- small cell lung cancer). In the past ten years, with the development of molecular medicine and the continuous emergence of targeted drugs, the treatment of non-small cell lung cancer has entered the era of individualized molecular targeted "precision" treatment from chemotherapy. The current individualized molecular targeted therap...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11C40B50/06
Inventor 阴层层车键为方鹏崔娜娜
Owner 广州漫瑞生物信息技术有限公司
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