High-throughput detection of ctDNA of hematopoietic and lymphatic tissue tumors

A lymphoid tissue and high-throughput technology, applied in the field of molecular diagnostics of hematological diseases, can solve the problems of lack of application of hematopoietic lymphoid tissue tumors and the lack of sequencing products for hematopoietic and lymphoid tissue tumors.

Pending Publication Date: 2020-04-17
珠海铂华生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the mainstream clinical applications of ctDNA liquid biopsy technology are still concentrated in a few high-in...

Method used

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  • High-throughput detection of ctDNA of hematopoietic and lymphatic tissue tumors
  • High-throughput detection of ctDNA of hematopoietic and lymphatic tissue tumors
  • High-throughput detection of ctDNA of hematopoietic and lymphatic tissue tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] The first step is to collect gene combination information by searching major authoritative databases at home and abroad (including COSMIC, NCBI, etc.) and combining authoritative journals and magazines. At the same time, an in-depth analysis is carried out on the data obtained by independent detection, gene classification is carried out through accurate clinical typing and gene pathway typing, data filtering of mutation sites is carried out through quality control data such as detection rate and Q20, and a batch of filtered leftovers are obtained. The final gene combination was merged with the gene combination of the first step, and finally a gene combination of 173 genes was obtained. In addition to covering almost all hematopoietic and lymphoid tissue tumor-related genes in current authoritative journals, this gene combination also covers hematopoietic and lymphoid tissue tumor types to prognosis related gene loci at all levels (including a number of inventors' new one...

Embodiment 2

[0127] 1. Extraction of sample DNA

[0128] A commercial ctDNA extraction kit ("QIAamp Circulating Nucleic Acid Kit" from Qiagen, product number 55114) was used to extract peripheral blood into a DNA solution, the quality of DNA 1730 / 230 was measured by Nonodrop, and the DNA concentration was detected by a Qubit instrument.

[0129] 2. DNA multiplex PCR library preparation

[0130] Use the DNA sample obtained in step 1 and the primer library obtained in Example 1 to build a library, use the "Ampliseq Library kit" of Lifetech Company, and operate according to the instructions. The specific steps are as follows:

[0131] 1) Prepare the DNA according to the following system:

[0132] components Reaction system (20μL) High Fidelity PCR Buffer 4 Primer library with 5 times concentration 4 ctDNA, 10ng Y Deionized water (12-Y) total capacity 20

[0133] Remarks: Y is the volume of liquid that needs to be added for 15ng DNA.

[0134] 2)...

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Abstract

The invention provides a gene combination comprising 173 myeloid tumor and lymphoid tissue tumor related genes, a primer combination for the gene combination, and applications of the gene combinationand the primer combination in high-throughput sequencing detection of myeloid tumors. According to the invention, hematopoietic and lymphatic tissue tumors can be comprehensively detected by using ctDNA.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis of blood diseases, and in particular relates to high-throughput sequencing detection of hematopoietic and lymphoid tissue tumors. Background technique [0002] Hematopoietic and lymphoid tissue tumors are a general term for tumors related to the hematopoietic system. In the latest classification of the World Health Organization (WHO), hematopoietic and lymphoid tissue tumors are divided into two categories: myeloid tumors and lymphoid tumors. It can be divided into multiple branches and subtypes. WHO's classification of hematopoietic and lymphoid tissue tumors is based on the four major disciplines of molecular biology, cytogenetics, cell morphology, and immunology. There are dozens of tumor subtypes, and the treatment methods and methods for different subtypes of tumors. Prognosis also varies. Therefore, accurate classification of hematopoietic and lymphoid tissue tumors is of great significa...

Claims

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Application Information

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IPC IPC(8): C40B50/06C40B40/06C12Q1/6886C12N15/11
CPCC40B50/06C40B40/06C12Q1/6886
Inventor 周剑峰何旭华肖敏张炜
Owner 珠海铂华生物工程有限公司
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