TEST SYSTEM AND METHOD FOR DETECTING A, B, D MUTATIONS OF NPM1 GENE FOR QUANTITATIVE DETERMINATION OF MINIMAL RESIDUAL DISEASE
	SIDOROVA YU V; SEVERINA N A; SUDARIKOV A B
	2023-09-25
专利权人	FEDERAL STATE BUDGETARY INST (FSBI-C)
申请日期	2023-09-25
专利号	RU2830545-C1
成果简介	NOVELTY - Described is a method for detecting A, B, D type NPM1 gene mutations in a test sample and determining MRD, involving: performing RT-PCR for primary analysis for detection of A, B, D type NPM1 gene mutations; setting of RT-PCR for MRD testing; determining MRD in percent, in the case of a sample containing DNA, according to the formula for calculating the allelic load (AL) of mutant allele 100*AO/(1+AO), where AO is the result of calculating the allelic ratio, wherein the allelic ratio (AO) is calculated by formula 5/2∆Ct, where ΔCt is the difference in amplification cycles for the mutant and wild allele, in the case of the sample containing RNA, according to the formula for calculating the relative expression of mutant allele 100/2ΔCt, where ΔCt is the difference in amplification cycles for the mutant and wild allele, wherein a set of reaction mixtures is used for RT-PCR, which includes four mixtures: mixture of oligonucleotides NPM1 W with sequences SEQ ID NO: 1, SEQ ID NO: 6, SEQ ID NO: 5; mixture of oligonucleotides NPM1 A with sequences SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 5; mixture of oligonucleotides NPM1 B with sequences SEQ ID NO: 3, SEQ ID NO: 6, SEQ ID NO: 5; mixture of oligonucleotides NPM1 D with sequences SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 5, intended for amplification of the analyzed DNA or RNA, which are specific for the wild-type NPM1 gene and for each type of NPM1 gene mutations A, B, D, after detecting the NPM1 mutation type A, B, D. Corresponding sets are disclosed. USE - Medicine; molecular biology. ADVANTAGE - Invention enables molecular genetic diagnosis of NPM1 gene mutations by a highly sensitive quantitative method based on allele-specific real-time PCR; analytical sensitivity of this method is 10−4–10−5 and allows detecting minimal residual disease (MRD) in biological material (bone marrow puncture and blood) of patients diagnosed with acute myeloid leukemia. 32 cl, 15 dwg, 15 tbl
IPC 分类号	C12Q-001/68
国家	俄罗斯
专业领域	生物科学
语种	英语
成果类型	专利
文献类型	科技成果
条目标识符	http://119.78.100.226:8889/handle/3KE4DYBR/20255
专题	中国科学院新疆生态与地理研究所
作者单位	FEDERAL STATE BUDGETARY INST (FSBI-C)
推荐引用方式 GB/T 7714	SIDOROVA YU V,SEVERINA N A,SUDARIKOV A B. TEST SYSTEM AND METHOD FOR DETECTING A, B, D MUTATIONS OF NPM1 GENE FOR QUANTITATIVE DETERMINATION OF MINIMAL RESIDUAL DISEASE. RU2830545-C1[P]. 2023.

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