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CHST2-SLC9A9-AS2 fusion gene as well as application and detection kit thereof

A CHST2-SLC9A9-AS2, fusion gene technology, applied in the field of biomedicine, to achieve the effects of good specificity, low false positives, safe and easy operation

Pending Publication Date: 2022-02-08
THE SECOND HOSPITAL OF DALIAN MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above two genes form a new fusion gene CHST2-SLC9A9-AS2 in ALL patients, which has not been reported in the literature so far

Method used

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  • CHST2-SLC9A9-AS2 fusion gene as well as application and detection kit thereof
  • CHST2-SLC9A9-AS2 fusion gene as well as application and detection kit thereof
  • CHST2-SLC9A9-AS2 fusion gene as well as application and detection kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1. Patients with acute lymphoblastic leukemia carry the new fusion gene CHST2-SLC9A9-AS2

[0039] (1) Applying bioinformatics technology to screen out fusion genes of patients with acute lymphoblastic leukemia and new fusion genes of CHST2-SLC9A9-AS2 that have not been reported in the literature.

[0040] (2) The new CHST2-SLC9A9-AS2 fusion gene is formed by splicing exon 1-2 of CHST2 gene and exon 2 of SLC9A9-AS2, and the specific base sequence is shown in SEQ ID NO.1.

[0041] (3) Using PCR and performing sanger sequencing on the PCR products, we confirmed that the patient with acute lymphoblastic leukemia carried the new fusion gene CHST2-SLC9A9-AS2, and the verification results are shown in figure 1 . figure 1 It is the CHST2-SLC9A9-AS2 fusion gene structure map and PCR, sanger verification. A: The translocation of the 3q24 region leads to a graphical display of the CHST2-SLC9A9-AS2 fusion gene; B: PCR verification of the fusion gene in the patient's prima...

Embodiment 2

[0044] Embodiment 2. Preparation of kit

[0045] 1. Design of specific primers and probes

[0046] According to the gene sequence (ABL1 gene sequence, CHST2 gene sequence, SLC9A9-AS2 gene sequence are all from the National Biotechnology Information Center nucleic acid database, ABL1 gene Entrez Gene ID25, gene reference sequence NM_005157.5; CHST2 gene Entrez Gene ID 9435, gene Reference sequence ENST00000309575.5; SLC9A9-AS2 gene Entrez Gene ID 106480356, gene reference sequence ENST00000490153.1) design specific probes and primers.

[0047] 2. Preparation of kit components

[0048] cDNA first-strand synthesis reagent: TIANScriptⅡRT Kit (TIANGEN Company), detection system PCR reaction solution: Pro Taq HS Premix Probe qPCR Kit (Acurrate Biology, AG11704), its main components include DNA polymerase, Mg 2+ , dNTPs, reverse transcriptase, DTT.

[0049] Primers and probes: including detection of CHST2-SLC9A9-AS2 fusion gene and internal reference ABL primers and probes corresp...

Embodiment 3

[0057] Example 3. The operation flow of this kit for detecting ALL patient specimens

[0058] 1. Take the anticoagulated blood samples of ALL patients submitted for inspection, and extract the total RNA in the blood: add 1ml of red blood cell lysate to a clean 1.5ml centrifuge tube, take 0.5ml of anticoagulated blood and mix well. Let stand at room temperature for 10 minutes; centrifuge at 5000rpm for 5min, discard the supernatant, and collect the cells at the bottom; add 0.5ml red blood cell lysate again, centrifuge at 5000rpm for 5min, discard the supernatant, and collect the cells at the bottom; add 1ml TRIzol to the cells, and pipette repeatedly until sedimentation Completely dissolve, let stand at room temperature for 5 minutes; add 0.2ml chloroform, shake evenly; centrifuge at 14000rpm 4°C for 10 minutes, absorb the supernatant layer and transfer to another new centrifuge tube; add an equal volume of isopropanol, mix well up and down, room temperature Let stand for 10min...

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Abstract

The invention provides a CHST2-SLC9A9-AS2 fusion gene as well as application and a detection kit thereof. The sequence of the fusion gene is as shown in SEQ ID NO. 1. The new fusion gene CHST2-SLC9A9-AS2 in acute lymphocytic leukemia is firstly found and identified, the abundance of the fusion gene is closely related to the disease progression stage, and the fusion gene can be used as a specific molecular marker of acute lymphocytic leukemia, so that a kit which can be clinically used for diagnosing and monitoring minimal residual disease (MRD) of a patient is developed. The kit uses Taqman probe real-time fluorescence PCR to carry out absolute quantification on the CHST2-SLC9A9-AS2 fusion gene in the body of an acute lymphocytic leukemia patient, the detection specificity is good, the sensitivity is high, the method is simple, convenient and efficient, the cost is low, and clinical diagnosis and treatment requirements can be met.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an acute lymphoblastic leukemia fusion gene and its application and detection kit. Background technique [0002] Acute lymphoblastic leukemia (ALL) is one of the most common hematological malignancies caused by the malignant proliferation of B or T lineage lymphoid progenitor cells. Due to the lack of normal peripheral blood cells, patients are clinically characterized by fever, recurrent infection, skin / mucosal hemorrhage, bone pain and arthralgia. Mediastinal mass and central nervous system are also often involved. The chemotherapy remission rate of children with ALL can reach 80-90%, but the prognosis of adult ALL is very poor, the overall survival rate is 35%-55%, and it is only 30% for adults over 60 years old. [0003] In order to assess the risk of recurrence and predict the clinical prognosis, monitoring minimal residual disease (MRD) is one of the commonly used clin...

Claims

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Application Information

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IPC IPC(8): C12N15/62C12N15/113C12Q1/6886C12Q1/686C12N15/11
CPCC12N9/13C12Y208/02C12N15/113C12Q1/6886C12Q1/686C12Q2600/166C07K2319/00C12Q2561/113C12Q2563/107
Inventor 张静静闫金松王海娜卢莹
Owner THE SECOND HOSPITAL OF DALIAN MEDICAL UNIV