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Gene probe composition and kit for detecting clone evolution of multiple myeloma

A technology of multiple myeloma and gene probes, applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problem of not being able to reflect the pathological process, and achieve the goal of improving detection ability and efficiency Effect

Active Publication Date: 2016-05-25
INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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Problems solved by technology

[0010] In summary, the five genes TP53, Rb1, CKS1B, CYLD and BIRC2 are closely related to the occurrence and development of multiple myeloma. At present, MM is mainly detected by a small number of single genes, and the results are far from reflecting the occurrence and development of multiple myeloma. complex pathological process

Method used

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  • Gene probe composition and kit for detecting clone evolution of multiple myeloma
  • Gene probe composition and kit for detecting clone evolution of multiple myeloma
  • Gene probe composition and kit for detecting clone evolution of multiple myeloma

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Embodiment 1

[0070] Embodiment 1: The preparation method of TP53 gene probe comprises the following steps:

[0071] 1. Primer design and clone screening: The TP53 gene is located at 17P13.1, the short arm of human chromosome 17. Search for all clones containing the TP53 gene in UCSCgenomebrowser, NCBICloneRegistry, and EnsemblGenomeBrowser databases, and use polymerase chain reaction to screen out the optimal clone containing the gene. The serial number is RP11-89D11 (as shown in Table 1).

[0072] 2. Cloning culture and identification: purchase clone RP11-89D11 (Invitrogen, USA), take 8 microliters of clone bacteria solution and add it to 5 milliliters of LB liquid medium containing chloramphenicol resistance, shake and activate for 12 hours at 37°C ; Then add all the bacterial liquid to 450 ml of LB liquid medium containing chloramphenicol resistance, and collect the bacterial liquid after shaking and culturing at 37°C for 12 hours. The upstream primer 5'-TGACACGCTTCCCTGGATTG-3' and the ...

Embodiment 2

[0101] Embodiment 2: the preparation method of TP53, Rb1, CKS1B, CYLD, BIRC2 gene probe composition, comprises the following steps

[0102] 1. Primer design and clone screening:

[0103]By searching UCSCgenomebrowser, NCBICloneRegistry, EnsemblGenomeBrowser and other databases, all clones containing TP53, Rb1, CKS1B, CYLD, BIRC2 genes were screened out by polymerase chain reaction, and the numbers were: RP11-89D11, RP11-305D15 , RP11-307C12, RP11-327F22, RP11-864G5 (as shown in Table 1).

[0104] 2. Cloning culture and identification: purchase the clone according to the clone number (Invitrogen, USA), take 8 microliters of the clone bacteria solution and add it to 5 milliliters of chloramphenicol-resistant LB liquid medium, shake and activate at 37°C for 12 hours Then all the bacterial liquid was added to 450 ml of LB liquid medium containing chloramphenicol resistance, and the bacterial liquid was collected after shaking and culturing at 37°C for 12 hours, and 5 target genes...

Embodiment 3

[0143] Embodiment 3: A kind of test kit (50 people) of detection multiple myeloma clonal evolution, composition is as follows:

[0144]

[0145] Among them, the fluorescent labeling probe group hybridization mixture includes TP53 gene probe, Rb1 gene probe, CKS1B gene probe, CYLD gene probe, BIRC2 gene probe, and the concentration of TP53 gene probe is: 4ng / μL, Rb1 gene probe Probe concentration: 4ng / μL, CKS1B gene probe concentration: 4ng / μL, CYLD gene probe concentration: 4ng / μL, BIRC2 gene probe concentration: 4ng / μL. The components of the hybridization mixture include 50% deionized formamide, 2×SSC and 0.1 g / ml dextran sulfate.

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Abstract

The invention relates to a gene probe composition and a kit for detecting clone evolution of multiple myeloma. The gene probe composition comprises a TP53 gene probe, an Rb1 gene probe, a CKS1B gene probe, a CYLD gene probe and a BIRC2 gene probe. The kit can be applied to the analysis of clone structures and clone heterogeneity of the multiple myeloma.

Description

technical field [0001] The invention relates to a gene probe composition and a kit, in particular to a gene probe composition and a kit for detecting multiple myeloma clone evolution. Background technique [0002] Multiple myeloma (Multiplemyeloma, MM) is a malignant blood system disease characterized by abnormal clonal proliferation of plasma cells and most of which are characterized by the production of monoclonal immunoglobulin (Immunoglobulin, Ig). The course of MM is clearly staged, from monoclonal gammopathy of undetermined significance (MGUS) to smoldering multiple myeloma (SMM), to symptomatic myeloma, to plasma cell leukemia (PCL) or Extramedullary plasmacytoma. Different disease courses have their different cytogenetic abnormalities. Therefore, MM has become a good model to study the clonal heterogeneity and clonal evolution of tumor cells. [0003] Although the clonal evolution of tumor cells has long been noticed, there is no fixed mature model for his researc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6841C12Q1/6886C12Q2600/156C12Q2600/158C12Q2600/16C12Q2531/113C12Q2563/107
Inventor 邱录贵安刚秦小琪刘兰婷胡林萍程涛
Owner INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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