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A group of thyroid cancer markers and their applications

A technology of thyroid cancer and markers, applied in the direction of recombinant DNA technology, microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of treatment plan selection or prognosis evaluation without guiding significance, molecular information cannot be obtained, etc.

Active Publication Date: 2020-08-25
SHANGHAI BIOTECAN PHARMA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the genes used in the above kits are mainly genes related to the molecular typing of thyroid cancer, and do not involve other thyroid cancer-related genes, so comprehensive molecular information cannot be obtained
[0009] "Application of Next-Generation Sequencing Technology in the Screening of Genetic Susceptibility Genes for Familial Nonmedullary Thyroid Cancer" mainly uses thyroid cancer molecular typing-related genes and thyroid cancer genetic-related genes, but does not involve medication or therapeutic markers , does not have guiding significance for the choice of treatment options or prognosis assessment

Method used

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  • A group of thyroid cancer markers and their applications
  • A group of thyroid cancer markers and their applications
  • A group of thyroid cancer markers and their applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0157] Example 1 gDNA extraction

[0158] Most of the detection samples of patients with thyroid disease are biopsy puncture samples, the sample size is small, and the quality of nucleic acid extraction is very important. In this example, the papillary thyroid carcinoma sample (tissue and blood) numbered BT1809140101LNDBE / TV is taken as an example to conduct genome analysis. DNA (gDNA) extraction was carried out using QIAGEN’s tissue and blood DNA extraction kits, the extraction steps were carried out according to the instructions, and the extracted nucleic acids were quality controlled using Qubit 3.0.

[0159] The extracted nucleic acid was digested and fragmented. The enzyme digestion system was shown in Table 1, and the enzyme digestion conditions were shown in Table 2. After the enzyme digestion reaction was completed, 40 μL of enzyme-free water was added to the enzyme digestion product to a total volume of 50 μL. Agilent 2100 was used to check the fragmented nucleic acid...

Embodiment 2

[0165] Embodiment 2 library construction

[0166] This embodiment builds a database with reference to the SureSelect XT Low Input System, and improves and optimizes the steps. The specific operations are as follows:

[0167] (1) End repair

[0168] Add enzyme-free water (Nuclease-free Water) to the fragmented sample obtained in Example 1 to make the volume up to 50 μL, prepare the end repair reaction system according to Table 3, mix it and place it on a PCR instrument at 20°C for 15 minutes, 72 Keep at ℃ for 15min, and store at 4℃ for later use.

[0169] Table 3 End repair reaction system

[0170] Reagent Dosage (μL) Buffer (End Repair-ATailing Buffer) 16 End Repair-ATailing Enzyme Mix 4 Fragmented samples 50 total capacity 70

[0171] (2) Joint connection

[0172] Prepare the adapter connection reaction system according to Table 2, mix it and place it on the PCR instrument for 30 minutes at 20°C, and store it at 4°C for later use; i...

Embodiment 3

[0192] Example 3 Target region capture of thyroid-related genes

[0193] This embodiment carries out target area capture with reference to SureSelect XT Low Input System, specific operation is as follows:

[0194] (1) capture hybridization

[0195] Transfer 1000ng of the pre-amplified library (including capture probes such as SEQ ID NO: 1 to 44) to a new centrifuge tube, dry it, and add 12 μL ddH 2 O dissolved, vortexed;

[0196] Add 5 μL of blocking solution (SureSelect XT HS and XT Low Input Blocker Mix), vortex to mix, transfer to a PCR plate, and place on a PCR machine for reaction, the conditions are shown in Table 9; when the program runs to step 3, pause , add 13 μL of the capture hybridization system shown in Table 10.

[0197] Table 9 target region capture reaction conditions

[0198]

[0199] Table 10 capture hybrid system

[0200] Reagent Dosage (μL) RNase Blocking Solution (25% RNase Block solution) 2 Library (Capture library 2 ...

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Abstract

The invention provides a group of thyroid cancer markers and an application thereof. The markers comprise thyroid cancer molecular typing related genes, targeted medication related genes, chemotherapymedication related genes, operation prompt related genes, prognosis related genes and thyroid cancer heredity related genes. According to the invention, aiming at all exon regions of 44 genes relatedto thyroid molecular typing, targeted medication, chemotherapy medication, operation prompt, prognosis and heredity, high-depth sequencing is carried out; meanwhile, various variation types like SNV / Indel and gene fusion of genes are analyzed; and molecular level information of thyroid nodules and thyroid cancer is deeply analyzed. Aiming at a capture probe designed for the 44 genes, 558 targeting target regions are covered; the probe comprises 3633 sequences with a total size of 254.096 Kbp; and a prepared kit is wide in coverage, high in cost performance and high in effectiveness, can provide a reference basis for further molecular typing, medication prompt, genetic risk assessment and the like of a thyroid patient, and is applicable to clinical popularization and application.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and relates to a group of thyroid cancer markers and applications thereof. Background technique [0002] Thyroid cancer (TC) is the most common endocrine gland malignancy, and the incidence rate has increased significantly in recent years. Among them, the incidence rate of women is higher than that of men, and the highest incidence age is 20-40 years old. According to the statistical report of the Ministry of Health of China in 2012, thyroid cancer has become the third largest female malignant tumor; the data of the National Cancer Center in 2017 shows that the incidence of thyroid cancer is increasing year by year, increasing at a rate of 6.2% every year, and urban women suffer from thyroid cancer. Cancer risk is higher. Thyroid cancer has become the most common malignant tumor in female patients in South Korea and Zhejiang Province of my country. It is estimated that by 2020, thyroid ca...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q1/6886C12Q2600/106C12Q2600/112C12Q2600/118C12Q2600/136C12Q2600/156C12Q2525/191C12Q2531/113C12Q2535/122
Inventor 魏利然林灵吴昊天杨广超刘静仪王永攀范梓伟许骋楼敬伟李紫薇
Owner SHANGHAI BIOTECAN PHARMA
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