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Reagent and kit for detecting fusion genes and applications of kit

A detection kit and gene fusion technology, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of reducing the recurrence rate and achieve the effect of predicting the progress of the disease or the possibility of recurrence

Pending Publication Date: 2019-04-12
BEIJING USCI MEDICAL LAB CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Trace amounts of leukemic cells remaining in decontaminated grafts are the main cause of higher relapse rates after autologous bone marrow transplantation, and molecular control of decontamination may reduce relapse rates

Method used

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  • Reagent and kit for detecting fusion genes and applications of kit
  • Reagent and kit for detecting fusion genes and applications of kit
  • Reagent and kit for detecting fusion genes and applications of kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] This embodiment comprises the steps:

[0039] (1) ARIDIB-ZNF384 fusion gene, DDX3X-MMLLT10 fusion gene, KMT2A-AF17 fusion gene, PATZ1-JAK2 fusion gene, CREBBP-ZNF384 fusion gene, EP300-ZNF384 fusion gene, RUNX1-MECOM fusion gene and ABL1 internal reference gene specificity Design of primers and probes;

[0040] (2) Design and preparation of standard products; establishment of real-time quantitative RT-PCR detection method and result processing.

[0041] (3) One-step real-time fluorescent quantitative RT-PCR to quickly detect the copy number of target fusion gene and internal reference gene ABL1 mRNA in leukemia bone marrow or peripheral blood samples, and use the correction ratio to provide reference for the efficacy detection and prognosis judgment of fusion gene-positive leukemia.

[0042] Using this method can simplify the traditional detection method. The specific detection method establishment and performance test results are as follows:

[0043] 1. Method establ...

Embodiment 2

[0115] 7 fusion gene positive samples (sample 1 is ARIDIB-ZNF384, sample 2 is DDX3X-MMLLT10, sample 3 is KMT2A-AF17, sample 4 is PATZ1-JAK2, sample 5 is CREBBP-ZNF384, sample 6 is EP300-ZNF384 and sample 7 is RUNX1-MECOM) common detection experiment process:

[0116] 1. For the extraction of total RNA from each of the 7 samples, the extraction method refers to the instructions of the Kangwei Reagent RNA Extraction Kit.

[0117] 2. Plasmid standard product dilution, take the ABL1 internal reference gene plasmid standard product, and use TE buffer to serially dilute to 1×10 6 copies / μL, 1×10 5 copies / μL, 1×10 4 copies / μL, 1×10 3 copies / μL and 1×10 2 copies / μL.

[0118] 3. Prepare a fluorescent quantitative RT-PCR system.

[0119] The RT-PCR system of the corresponding fusion gene and internal reference gene of each sample RNA, and the RT-PCR system of the ABL plasmid standard were respectively prepared in the following table 3.

[0120] table 3:

[0121]

[0122] 4. P...

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Abstract

The invention provides a reagent, a reagent and a kit for detecting fusion genes and applications of the kit. The reagent includes a detection reagent aiming at at least one of the following fusion genes: ARIDIB-ZNF384, DDX3X-MMLLT10, KMT2A-AF17 and PATZ1-JAK2. The present invention first discovered the association between four fusion genes and leukemia, so that the related detection reagent designed based on any one or more of the four fusion genes not only can be used for detecting the expression quantity of the corresponding fusion genes, but also can be used for monitoring changes of the expression quantity of corresponding fusion genes in leukemia patients so as to monitor the corresponding conditions. Moreover, based on the above-mentioned fusion genes, according to the actual monitoring needs, the product can also be used together with the previously reported leukemia-related marker genes, thereby relatively and comprehensively predicting the progression condition of leukemia orthe recurrence possibility.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a reagent, kit and application for detecting fusion genes. Background technique [0002] With the improvement of chemotherapy regimens for acute leukemia and the progress of hematopoietic stem cell transplantation, the complete remission (CR) rate of acute leukemia has increased significantly. However, there are still many CR patients who relapse within a few years, mainly due to the residual 10 6 ~10 9 leukemia cells, known as minimal residual disease (MRD). Some scholars believe that MRD is the root cause of leukemia relapse during hematological CR and even sustained complete remission (CRR). How to diagnose MRD early and accurately is the prerequisite for the prevention and treatment of acute leukemia relapse. [0003] Existing data show that there is a high correlation between MRD detection with fusion genes as target genes and disease-free survival, which has important clin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158
Inventor 张淑丽李富威方楠王建伟伍启熹刘倩刘珂弟唐宇
Owner BEIJING USCI MEDICAL LAB CO LTD
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