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Methods for rapid and sensitive detection of hotspot mutations

一种身体、样品的技术，应用在遗传和生物化学分析领域，能够解决不适用临床用途、误导疗法等问题

Active Publication Date: 2017-05-31
DUKE UNIV
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Problems solved by technology

These limitations have significant diagnostic and prognostic inferences and can produce misguided therapies, making them unsuitable for clinical use

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  • Methods for rapid and sensitive detection of hotspot mutations
  • Methods for rapid and sensitive detection of hotspot mutations
  • Methods for rapid and sensitive detection of hotspot mutations

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[0038] TERT promoter and IDH1 / 2 hotspot mutation AS LNA q-PCR analysis:

[0039] A. Background :

[0040] Allele-specific PCR is a form of DNA template amplification for the selective amplification of templates containing the variant of interest, and is thus a method for SNP genotyping 18 . Most methods rely on identifying primers with higher complementarity to target sequences with the genotype of interest. This is most commonly done by forcing the 3' of the primer to be at the mutation position and only complementary to the target variant or wild-type nucleotide. In this way, PCR efficiency decreases when primers bind to non-target alleles, imparting selective amplification ( Image 6 ).

[0041] The major advance in this region has been the use of candidate nucleic acids at the 3' end of these allele-specific primers called locked nucleic acids (LNAs). LNAs are nucleic acid analogs with a methylene bridge between the 2'-O and 4'-C of the nucleic acid, which creates...

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Abstract

Methods that rapidly, sensitively, and specifically detect mutations in IDH1 / 2 and the TERT promoter employ amplification of particular portions of the genes that experience frequent and exquisitely localized mutations. The ability to distinguish between sequences that differ only by one nucleotide and which may be present in very low ratios is essential for such an assay.

Description

technical field [0001] The present invention relates to the fields of genetic and biochemical analysis. Specifically, it concerns the analysis of neoplastic samples and their components. Background technique [0002] Malignant glioma is the most common primary central nervous system (CNS) malignancy in adults, causing >14,000 deaths in the US in 2012 2 . The World Health Organization (WHO) has established a number of histological and clinical guidelines for classifying gliomas into various subtypes and grading them from I to IV (indicating their degree of malignancy). Diffuse glioma (WHO grade II-IV), which includes astrocytoma, oligodendroglioma, oligoastrocytoma, and glioblastoma (GBM) 3 , is of particular clinical importance as it accounts for 80% of all primary malignant brain tumors. These tumors are diffusely infiltrating, which makes curative surgical resection impossible. In addition, grade II–III diffuse gliomas also have the capacity to progress to higher W...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12P19/34C12N15/10
CPCC12Q2600/16C12Q2600/166C12Q2600/156C12Q1/6886C12Q2521/101C12Q2527/101C12Q2531/113C12Q2525/117C12Q2600/158
Inventor H·严Y·何R·杨B·H·迪普拉斯L·汉森D·比格纳
Owner DUKE UNIV
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