BCR/ABL fusion gene mRNA fluorescence quantitative PCR detecting kit

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...

AI Technical Summary

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 labeled separately. The probe will affect the PCR amplification during the binding process, affecting the optimization of the PCR system and reaction conditions, and the synthesis of the probe and the double labeling of the probe are expensive, which brings great economic pressure to the patient

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