BCR/ABL fusion gene mRNA fluorescence quantitative PCR detecting kit

A detection kit and fusion gene technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problems of high price, affecting the optimization of PCR system and reaction conditions, etc., and achieve high specificity, high sensitivity, and fluorescence signal enhancement Effect

Inactive Publication Date: 2008-04-30
冯文莉
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Problems solved by technology

Domestic commercial fluorescent quantitative PCR kits are also mostly developed using Taqman probe technology, but this technology requires the synthesis of 1 to 2 fluorescent probes separately in addition to the amplification primers, and the fluorescent groups and quenching groups must be la

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  • BCR/ABL fusion gene mRNA fluorescence quantitative PCR detecting kit

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[0028]The BCR / ABL fusion gene mRNA fluorescent quantitative PCR detection kit includes lymphocyte separation liquid, RNA extraction liquid A, RNA extraction liquid B, Oligo(dT) 12-18 , RT reaction solution, reverse transcriptase, quantitative PCR reaction solution, standard substance, reference substance, DEPC water, quantitative PCR reaction solution contains PCR buffer, MgCl 2 , dNTPs, sterile deionized water, labeled primers and unlabeled primers. RNA extraction solution A is Trizol; RNA extraction solution B is 75% (v / v) ethanol prepared with DEPC water; Oligo(dT) 12-18 The concentration is 0.5μg / μl; the RT reaction solution contains 2.8×RT buffer, 22.2U / μl RNasin, 1.6mM dNTPs, DEPC water; the reverse transcriptase is 200u / μl M-MLV; the quantitative PCR reaction solution contains 1.3 ×PCR buffer, 2.7mM MgCl 2 , 0.33mM dNTPs, 0.33pmol / μl labeled primer (self-quenching primer), 0.33pmol / μl unlabeled primer, 0.17U / μl Taq DNA polymerase, sterile deionized water; the standard...

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Abstract

The invention relates to a BCR/ABL fusion gene (P210bcr/abl) mRNA fluorescence quantitative PCR measurement detection reagent box which comprises lymphocyte segregating liquid, RNA extracting liquid A, RNA extracting liquid B, Oligo (dT) 12-18, RT reaction fluid, reverse transcriptase, quantitative PCR reaction fluid, standard sample, comparison sample, and DEPC water; wherein, a marked primer and a non-marked primer are contained in the quantitative PCR reaction fluid. The reagent box can accurately detect the BCR/ABL fusion gene (P210) mRNA level in the specimen to be detected by extracting the total RNA of medulla ossium or peripheral blood, obtaining cDNA through reverse transcriptase and being combined with a real-time fluorescence quantitative PCR measurement detection technology. The reagent box adopts the latest self-quenched probe technology, thereby having the advantages that the repetitiveness is good, the sensitivity is high, the cost is low, and the invention can be applied to the dynamic monitoring of chronic myelocytic leukemia diagnosis, curative effect observation, prognosis and micro residual leukemia (MRD).

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to BCR / ABL fusion gene (P210 bcr / abl )mRNA self-quenching probe fluorescent quantitative PCR detection kit is a kind of cDNA obtained by extracting total RNA from clinical bone marrow and reverse transcription, combined with real-time fluorescent quantitative PCR detection technology, it can accurately quantify the BCR / ABL fusion gene (P210 bcr / abl ) mRNA expression kit, which provides a basis for the diagnosis, curative effect observation and prognosis judgment of chronic myelogenous leukemia. Background technique [0002] Chronic myelogenous leukemia (CML) is a malignant blood disease originating from abnormal clones of bone marrow multipotent hematopoietic stem cells. It has a characteristic Philadelphia chromosome (Ph') with a typical karyotype of t(9;22)(q34;q11), a chromosomal translocation that results in the fusion of two normal genes, BCR and ABL, to form a neoplasti...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 彭辉王小中刘钉宾曾建明
Owner 冯文莉
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