Method for detecting multi-site mutation of free DNA of lung tumor plasma
A technology for multi-site mutations and lung tumors, applied in recombinant DNA technology, DNA/RNA fragments, biochemical equipment and methods, etc., can solve the problems of poor specificity and low sensitivity, and achieve rapid detection, good repeatability, and improved Effects of Sensitivity and Specificity
Pending Publication Date: 2022-02-01
深圳思凝一云科技有限公司
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- Abstract
- Description
- Claims
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Problems solved by technology
The existing ctDNA detection methods mainly have the disadvantages of low sensitivity and poor specificity
Method used
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Experimental program
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Effect test
Embodiment approach I
[0046] Fragment the standard gDNA, add adapters, and PCR enrichment to prepare the pre-library; then proceed to the subsequent steps (1)-(3)
Embodiment approach II
[0048] Fragmentation, addition of adapters, and PCR enrichment were performed on tissue DNA extracted from FFPE samples to prepare a pre-library;
[0049] Then follow up steps (1)-(3)
Embodiment approach III
[0051] Prepare a pre-library by adding adapters to standard cfDNA and PCR enrichment;
[0052] Then follow up steps (1)-(3)
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Abstract
The invention provides a method for detectingmulti-site mutation of free DNA of lung tumor plasma. The method is characterized in that a specific primer, a specific PNA probe and a capture technology are adopted to improve ctDNA detection and specificity, so that trace ctDNA can be detected. The invention provides primers, probes and a detection method for gene variation detection in a peripheral blood sample of a lung space-occupying lesion patient. The method has the advantages of high sensitivity, strong specificity, good repeatability, simplicity in operation, rapidness in detection, safety and the like, and has an auxiliary effect on diagnosis of lung cancer.
Description
technical field [0001] The invention relates to a method for detecting DNA multi-site mutations, in particular to a method for detecting free DNA multi-site mutations in retired lung tumor plasma. Background technique [0002] With the development of next-generation sequencing technology, "liquid biopsy", as a new diagnostic technology, has developed rapidly in recent years due to its non-invasive and real-time dynamic characteristics, and is becoming a new hot spot in the field of tumor precision medicine. Among them, gene detection of body fluid samples represented by plasma has attracted the most attention. This method obtains free nucleic acid or cells from peripheral blood to find tumor information, and then provides a basis for early diagnosis. Lung cancer ranks first among all cancers in terms of morbidity and mortality in China, and early diagnosis and screening are imminent. [0003] At present, the technical difficulties in accurately sequencing ctDNA and realizin...
Claims
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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6806C12Q1/6837C12Q1/6827C12N15/11
CPCC12Q1/6886C12Q1/6806C12Q1/6837C12Q1/6827C12Q2600/156C12Q2525/191C12Q2531/113C12Q2539/105C12Q2563/143C12Q2563/149
Inventor 李小花刘朝煜姚旭梅
Owner 深圳思凝一云科技有限公司
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