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Leukemia MEF2D gene disruption probe detection kit

A technology of gene breakage and kit, which is applied in recombinant DNA technology, microbial measurement/testing, DNA/RNA fragments, etc., can solve the problems of expensive detection, complicated operation, and long time, so as to achieve optimal treatment and prognosis evaluation, The effect of strong fluorescent signal and high accuracy

Active Publication Date: 2019-08-06
BEIJING CHILDRENS HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that RNA sequencing has extremely high requirements on sample quality. If the quality control of extracted RNA is not up to standard, subsequent experiments will not be possible.
Therefore, in clinical applications, it is difficult to apply this technology independently, and the positive results of the analysis must be verified by combining PCR methods.
Due to the complex operation and long time of this method, it is difficult to meet the needs of rapid clinical diagnosis
In addition, the detection cost of this method is expensive, which is a bottleneck problem that limits its popularization and application.

Method used

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  • Leukemia MEF2D gene disruption probe detection kit
  • Leukemia MEF2D gene disruption probe detection kit
  • Leukemia MEF2D gene disruption probe detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Embodiment 1, MEF2D gene fragmentation detection probe, preparation and use method of kit

[0047] Technical thinking of the present invention is:

[0048] Fluorescence in situ hybridization is a method that uses a probe labeled with fluorescein to specifically combine with chromosomes and (or) gene loci, and observes the type of fluorescent signal through a fluorescence microscope to detect changes in chromosomes and corresponding genes. It is safe and economical. , fast, high sensitivity, strong detection signal, high hybridization specificity, and can display multiple colors at the same time, etc., and makes up for the defects that traditional methods cannot diagnose interphase cells, complex karyotype cells, and chromosomal microdeletions. At the same time, fluorescence in situ hybridization technology is applied to paraffin-embedded samples for retrospective research, which greatly reduces the requirements for research samples. Based on the rapid development of fl...

Embodiment 2

[0066] Example 2, Practical application of MEF2D gene breakage detection probes and kits

[0067] In this embodiment, the bone marrow of 40 children with clinically confirmed ALL (30 of the 40 children were B-ALL, and 10 were T-ALL; their guardians were informed and agreed) was used as the sample to be tested, and the bone marrow droplet sample was prepared. , use the probe kit in Example 1 to detect whether the MEF2D gene is broken or not, and refer to the method in Step 3 of Example 1 for details.

[0068] The results showed that 2 of the 30 bone marrow cell droplet specimens from clinically diagnosed B-ALL patients met the diagnostic criteria of fluorescence in situ hybridization and were judged to be positive, while the remaining 28 cases were diagnosed as negative. In the bone marrow cell droplet samples of 10 clinically diagnosed T-ALL patients, no gene break signal was detected, and they were judged as negative. figure 2 It is a negative control case of children with ...

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Abstract

The invention discloses a leukemia MEF2D gene disruption probe detection kit. The present invention provides a fluorescent in-situ hybridization polyclonal separating probe for detecting chromosomal MEF2D gene disruption, which is composed of by two BAC clone fragments (RP11-964F7 and RP11-139I14) located on the centromere side of the chromosomal MEF2D gene and two BAC clone fragments (RP11-214H6and RP11-1047J23) located on the telomere side of the chromosomal MEF2D gene. The kit utilizes a FISH technology to detect leukemia associated with MEF2D gene disruption, and performs individualized treatment for patients. The probe of the invention can comprehensively detect all translocations involving the MEF2D gene, finds new translocations, has high application accuracy, high specificity, high success rate, strong fluorescence signal and simple operation, and can assist in the optimization of the treatment and prognosis of leukemia associated with the MEF2D gene disruption.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a leukemia MEF2D gene breakage probe detection kit. Background technique [0002] B-cell acute lymphoblastic leukemia (ALL) is the most common malignant tumor in childhood. Molecular typing based on genetic abnormalities (gene fusion, aneuploidy) can help guide clinical diagnosis, risk stratification, and targeted therapy, greatly improving the cure rate of B-ALL. However, there are still about 20% of children with relapse. The reason is that there are insufficient risk grading indicators, lack of targeted therapy, and insufficient research on the pathogenesis. Fusion gene is one of the important causes of childhood B-ALL and is associated with ALL risk classification and targeted therapy [1,2] . Therefore, discovering new fusion genes and elucidating their mechanism of action are of great significance for revealing the mechanism of leukemia development, as well as B-ALL risk stra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6841C12N15/11
CPCC12Q1/6886C12Q1/6841C12Q2600/118C12Q2600/156C12Q2563/107C12Q2563/173
Inventor 高超岳志霞刘曙光田硕郑胡镛张瑞东陈绍宇
Owner BEIJING CHILDRENS HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV
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