Kit for quantitatively detecting AML1/ETO mRNA level

A quantitative detection and kit technology, which is applied in the field of kits for quantitative detection of AML1/ETO mRNA levels, achieves the effects of strong practicability, cost reduction and cost reduction

Inactive Publication Date: 2010-01-13
PEOPLES HOSPITAL PEKING UNIV
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Problems solved by technology

(3) MRD monitoring: The current treatment for AML1 / ETO(+) AML patients includes chemotherapy and hematopoietic stem cell transplantation, which can make most patients achieve genetic complete remission, and subsequent MRD monitoring can only rely on the current Recognized as the most sensitive RQ-PCR technology

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  • Kit for quantitatively detecting AML1/ETO mRNA level
  • Kit for quantitatively detecting AML1/ETO mRNA level

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Embodiment 1

[0024] Embodiment 1, the preparation of the kit of quantitative detection AML1 / ETO mRNA level

[0025] 1. Design of primers and probes in the kit for quantitative detection of AML1 / ETO mRNA levels

[0026] AML1 / ETO real-time quantitative PCR system is as follows:

[0027] (1) Upstream primer (located at exon 5 of AML1): 5'-CACCTACCACAGAGCCATCAAA-3' (sequence 1 in the sequence listing), with a final concentration of 0.3 μM;

[0028] (2) Downstream primer (located at ETO exon 2): 5'-ATCCACAGGTGAGTCTGGCATT-3' (sequence 2 in the sequence listing), the final concentration is 0.3 μM;

[0029] (3) TaqMan probe (located in ETO exon 2): 5'-AACCTCGAAATCGTACTGAGAAGCACTCCA-3' (sequence 3 in the sequence listing), the final concentration is 0.2 μM;

[0030] (4) Master Mix for fluorescent PCR (purchased from ABI, USA).

[0031] The internal reference gene real-time quantitative PCR system is as follows:

[0032] (1) Upstream primer (located in ABL exon 2):

[0033] 5'-TGGAGATAACACTCTAA...

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Abstract

The invention discloses a kit for quantitatively detecting an AML1 / ETO mRNA level. The kit comprises a standard product which is used for manufacturing a standard curve, an inner reference gene real-time quantitative PCR system and an AML1 / ETO real-time quantitative PCR system. The kit can accurately, quickly and quantitatively detect the AML1 / ETO mRNA level, is used for diagnosing acute myelogenous leukemia and monitoring minimal residual diseases in a treatment process, and provides an important molecular basis for accurate diagnosis of clinical diseases, determination of a treatment proposal, curative effect evaluation and prognosis.

Description

technical field [0001] The invention relates to a kit for quantitatively detecting AML1 / ETO mRNA level. Background technique [0002] Acute myeloid leukemia (AML) chromosomal translocation t(8; 21) (q22; q22) is manifested at the molecular level as the fusion gene AML1 / ETO of AML1 gene and ETO gene, and about 8-12% of adults and children are genetically This fusion gene is contained in episodic AML, and about 40% of M2 AML patients contain this fusion gene. In addition, there are also a few cases reported in M0, M1 and M4 AML patients. The AML1 gene encodes the α-subunit of the heterodimeric transcription factor CBF, which is a key factor in the development of the hematopoietic system, and the ETO protein is a transcriptional co-repressor. At present, the mechanism of AML1 / ETO triggering AML is still not very clear. The AML1 / ETO fusion gene type is single, the breakpoints on AML1 are all on intron 5, and the breakpoints on ETO are all upstream of exon 2, so a set of primer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 刘艳荣秦亚溱主鸿鹄李金兰李玲娣
Owner PEOPLES HOSPITAL PEKING UNIV
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