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Leukemia ZNF384 gene fragmentation probe disruption kit

A technology of gene breakage and kit, which is applied in the direction of recombinant DNA technology, microbial detection/testing, DNA/RNA fragments, etc., can solve the problems of high cost of 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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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 ZNF384 gene fragmentation probe disruption kit
  • Leukemia ZNF384 gene fragmentation probe disruption kit
  • Leukemia ZNF384 gene fragmentation probe disruption kit

Examples

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Comparison scheme
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Embodiment 1

[0047] Embodiment 1, ZNF384 gene fragmentation detection probe, preparation and use method of kit

[0048] Technical thinking of the present invention is:

[0049] 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 f...

Embodiment 2

[0067] Example 2, Practical application of ZNF384 gene breakage detection probe and kit

[0068] 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. , using the probes and kits in Example 1 to detect whether the ZNF384 gene is broken or not. For details, refer to the method in Step 3 of Example 1.

[0069] The results showed that among the 30 bone marrow cell droplet specimens from clinically diagnosed B-ALL patients, 2 cases met the diagnostic criteria of fluorescence in situ hybridization and were judged 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 child...

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Abstract

The invention discloses a leukemia ZNF384 gene fragmentation probe disruption kit. The present invention provides a fluorescent in-situ hybridization polyclonal separating probe for detecting chromosomal ZNF384 gene disruption, which is composed of two BAC clone fragments (CTD-2640D15 and RP11-256J20) located on the centromere side of the chromosome ZNF384 gene and two BAC clone fragments (RP11-687D2 and RP11-773K10) located on the telomere side of the ZNF384 gene. The kit utilizes a FISH technology to detect leukemia associated with ZNF384 gene disruption, and performs individualized treatment for patients. The probe of the invention can comprehensively detect all translocations involving the ZNF384 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 ZNF384 gene disruption.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a leukemia ZNF384 gene break probe detection kit. Background technique [0002] B-cell acute lymphoblastic leukemia (B-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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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/142
Inventor 岳志霞高超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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