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Kit for quantitatively detecting BCR/ABL mRNA level

A quantitative detection and kit technology, which is applied in the field of kits for quantitative detection of BCR/ABL mRNA levels, can solve problems such as poor prognosis, and achieve the effects of cost reduction, cost reduction, and high sensitivity

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

(2) Prognosis: The prognosis of Ph(+) ALL patients is very poor, and hematopoietic stem cell transplantation is necessary for long-term survival
(3) MRD monitoring: At present, there are effective methods such as hematopoietic stem cells and ABL tyrosine kinase inhibitors (such as Glivec) for the treatment of CML and Ph(+) ALL patients, which can make quite a lot of patients acquire genetic MRD monitoring can only rely on the currently recognized most sensitive RQ-PCR technology

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  • Kit for quantitatively detecting BCR/ABL mRNA level
  • Kit for quantitatively detecting BCR/ABL mRNA level

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Embodiment 1

[0025] Embodiment 1, the preparation of the kit of quantitative detection BCR / ABL mRNA level

[0026] 1. Design of primers and probes in the kit for quantitative detection of BCR / ABL mRNA levels

[0027]The composition of the M-type BCR / ABL real-time quantitative PCR system is as follows:

[0028] (1) Upstream primer (located at BCR exon 13): 5'-CCGCTGACCATCAATAAGGAA-3' (sequence 1 in the sequence listing), with a final concentration of 0.3 μM;

[0029] (2) Downstream primer (located at ABL exon 2): 5'-CTCAGACCCTGAGGCTCAAAGT-3' (sequence 2 in the sequence listing), with a final concentration of 0.3 μM;

[0030] (3) TaqMan probe (located in ABL exon 2): 5'-AGCCCTTCAGCGGCCAGTAGCATCT-3' (sequence 3 in the sequence listing), the final concentration is 0.2 μM;

[0031] (4) Master Mix for fluorescent PCR (purchased from ABI, USA).

[0032] The composition of the m-type BCR / ABL real-time quantitative PCR system is as follows:

[0033] (1) Upstream primer (located at BCR exon 1): ...

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Abstract

The invention discloses a kit for quantitatively detecting a BCR / ABL 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 at least one of the following three real-time quantitative PCR systems: an M-type BCR / ABL real-time quantitative PCR system, m-type BCR / ABL real-time quantitative PCR system and a mu-type BCR / ABL real-time quantitative PCR system. The kit can accurately, quickly and quantitatively detect various BCR / ABL mRNA levels, is used for diagnosing chronic myelogenous leukemia and acute lymphoblastic leukemia expressed by BCR / ABL 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 BCR / ABL mRNA level. Background technique [0002] 95% of chronic myelogenous leukemia (CML), 25-30% of adult acute lymphoblastic leukemia (ALL) and 2-5% of children with ALL have a characteristic Ph chromosome, interacted by t(9;22) Ectopic formation, at the molecular level, the BCR gene on chromosome 22 and the ABL gene on chromosome 9 form a BCR / ABL fusion gene. The breakpoints of ABL gene were concentrated in the upstream of exon 1b, between 1b and 1a or upstream of a2, and the ABL fusion part was all a2 at the mRNA level. The breakpoints on the BCR gene are mainly concentrated in the following three regions: the 5.8kb region spanning exons 12-16, namely the M-BCR region; the 55kb region on the first intron, namely the m-BCR region and the 19th intron On the intron, that is, the μ-BCR region. Due to the different breakpoints on the BCR gene, BCR / ABL fusion proteins with different molecul...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q2600/156C12Q1/6886C12Q2600/158
Inventor 秦亚溱刘艳荣主鸿鹄李金兰李玲娣陈珊珊
Owner PEOPLES HOSPITAL PEKING UNIV
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