Accurate and massively parallel quantification of nucleic acid

An oligonucleotide and labeling nucleotide technology, applied in the field of a method and a kit containing probes, can solve the problem of high cost of sequencing library preparation, waste of unrelated locus sequencing work, limited interpretation data statistical selection, etc. problem, to achieve high analytical specificity

Pending Publication Date: 2020-04-17
EAWAG SWISS FEDERAL INST OF AQUATIC SCI & TECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, current methods suffer from high cost of sequencing library preparation and waste of sequencing effort on sequencing non-related gene targets
For example, in cancer-associated liquid biopsies, non-targeted approaches lead to wasted sequencing of non-relevant loci in oncology
Untargeted sampling of epitope loci in fetal diagnostics greatly limits statistical options for interpreting data

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  • Accurate and massively parallel quantification of nucleic acid
  • Accurate and massively parallel quantification of nucleic acid
  • Accurate and massively parallel quantification of nucleic acid

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Embodiment Construction

[0039] The following detailed description illustrates embodiments of the invention and how to achieve it. Although a few modes for carrying out the invention have been disclosed, those skilled in the art will recognize that other embodiments for carrying out or practicing the invention are also possible.

[0040] In a first embodiment, the present invention provides a method for high-throughput detection of one or more target nucleotide sequences in a plurality of samples, the method comprising the steps of:

[0041] (i) providing for each target nucleotide sequence in each sample: a first probe, a second probe, and a bridge oligonucleotide, wherein

[0042] The first probe includes an optional 5' phosphate starting at the 5' end of the molecule, a first bridge oligonucleotide specific sequence, an optional first universal sequence, an optional first sequence barcode, and a first target-specific portion at the 3' end of the probe; and wherein

[0043] The second probe includ...

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Abstract

The present invention disclosure relates to a next generation DNA sequencing method and use for accurate and massively parallel quantification of one or more nucleic acid targets. More particularly, the invention is related to the method and a kit comprising probes for detecting and quantifying genetic targets in complex DNA pools primarily used for genetic target and variant detection in human and animal populations and environmental samples. Furthermore, the invention finds particular application in the field of detection of disease-causing genetic alterations in samples obtained from humanbody, including without limiting biopsies, saliva and other secretions, exhaled moisture extracts, tissue, blood plasma (liquid biopsies) or the like. The invention includes one or more target-specific nucleic acid probes per genetic target (left probe and right probe) and a bridge oligo.

Description

technical field [0001] The present disclosure relates to next generation DNA sequencing methods and uses for precise and massively parallel quantification of one or more nucleic acid targets. More specifically, the present invention relates to methods and kits containing probes for the detection and quantification of gene targets in complex DNA libraries primarily used in human and animal populations as well as in environmental samples. Gene target and variant detection. In addition, the present invention is useful in the detection of disease-causing genes in samples obtained from the human body, including but not limited to biopsy samples, saliva and other secretions, exhaled water extracts, tissues, plasma (liquid biopsy samples), etc. Special applications are found in changing fields. The invention includes one or more target-specific nucleic acid probes (left and right probes) and bridge oligonucleotides for each gene target. Background technique [0002] As technique...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/6855C12Q1/6865C12Q1/6869
CPCC12Q1/6806C12Q1/6855C12Q1/6865C12Q1/6869C12Q2521/531C12Q2525/143C12Q2525/161C12Q2525/179C12Q2525/307C12Q2531/125C12Q2535/122C12Q2537/159C12Q2537/162C12Q2563/179
Inventor 马努·塔米宁蒂莫西·朱利安珍妮·斯帕克莉亚·卡杜夫汉娜·希夫
Owner EAWAG SWISS FEDERAL INST OF AQUATIC SCI & TECH
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