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Application of chromosome instable mutation to preparing agent or kit for diagnosis, evaluation and prognosis of multiple myeloma

A multiple myeloma, chromosome technology, applied in the field of biotechnology and medical diagnosis, can solve the lack of multiple myeloma chromosome distribution characteristics and other problems, to achieve high sensitivity, simple operation, high accuracy

Pending Publication Date: 2019-10-11
SUZHOU HONGYUAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, there is a clinical method using in situ fluorescence hybridization to detect instability in some regions of chromosomes, but it lacks the distribution characteristics of chromosome instability at the entire genome level in patients with multiple myeloma, which has more comprehensively guided multiple myeloma diagnosis and prognosis

Method used

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  • Application of chromosome instable mutation to preparing agent or kit for diagnosis, evaluation and prognosis of multiple myeloma
  • Application of chromosome instable mutation to preparing agent or kit for diagnosis, evaluation and prognosis of multiple myeloma
  • Application of chromosome instable mutation to preparing agent or kit for diagnosis, evaluation and prognosis of multiple myeloma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation of library adapters

[0043] Linker sequences synthesized by third-party companies:

[0044] Linker-1 (SEQ ID NO.1):

[0045] 5'-CGTGATAGATCGGAAGAGCACACGTCTGAACTCCAGTC-3';

[0046] Linker-2 (SEQ ID NO.2):

[0047] 5'-ACACTCTTTCCCTACACGACGCTCTTCCGATCTATCACGT-3'.

[0048] The 5' end of Adapter-1 is modified by phosphorylation, and the 3' end of Adapter-2 is modified by phosphorothioation between G and T.

[0049] Use the annealing buffer (Annealing buffer) to dilute the dry powder of adapter-1 and adapter-2 to 20 μM respectively, mix the two dilutions in equal proportions, put the mixture in the PCR instrument, and run the program as shown in Table 1:

[0050] Table 1

[0051]

[0052]

[0053] The product was subjected to agarose electrophoresis, and the size range of the electrophoresis band was between 80-100bp.

Embodiment 2

[0055] Materials used:

[0056] human genomic DNA

[0057] DNA fragmentation reagent (Enzymatics)

[0058] DNA end repair, adding A, adding adapter enzyme reaction system (NEB)

[0059] PCR amplification reaction system (Kapa Biosystems)

[0060] Purification magnetic beads (Beckman)

[0061] Steps:

[0062] 1. Genome Fragmentation

[0063] Take 20ng of genomic DNA, use the enzyme digestion reaction system, configure the reaction system according to Table 2, and perform the reaction according to Table 3.

[0064] For cell-free DNA, such as cfDNA in blood, it does not need to undergo genome fragmentation and can directly enter the next step.

[0065] Table 2

[0066] DNA fragmentation reaction system Genomic DNA xμL interrupt enzyme 3μL Buffer 7μL nucleic acid free water 25-x μL total capacity 35μL

[0067] Shake to mix, centrifuge (avoid air bubbles) and run the following program on the PCR instrument:

[0068] table ...

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Abstract

The invention relates to the field of biotechnology and medical diagnosis, in particular to application of chromosome instable regions to preparing an agent or kit for the diagnosis of multiple myeloma. There are seven chromosome instable regions 1p, 1q, 6q, 11q13.3, 13q, 14q and 17p. Samples of clinically confirmed multiple myeloma patients and healthy people are collected, the specific chromosome instability distribution characteristics of the multiple myeloma are expounded by comparing chromosome abnormity conditions of the tumor patients and the contrast through whole-genome sequencing, the seven common chromosome instable regions of the multiple myeloma patients are disclosed, and the total carrying rate is up to 89.3%, which has great significance in the clinical diagnosis, treatment, monitoring, prognosis and evaluation of the multiple myeloma and provides the scientific basis for conducting early diagnosis next and formulating an individualized treatment scheme.

Description

technical field [0001] The present invention relates to the technical fields of biotechnology and medical diagnosis, specifically, a group of chromosomal instability mutations related to multiple myeloma, and the use of these unstable mutations in preparing reagents or reagents for diagnosing multiple myeloma and evaluating prognosis application in the box. Background technique [0002] Multiple myeloma (Multiple Myeloma, MM) is a malignant tumor characterized by the accumulation of plasma cells in the bone marrow, which can lead to bone destruction and bone marrow failure. According to statistics, MM accounts for 1% of all cancers and 10% of hematologic malignancies. In 2018, there were 30,770 new MM cases and 12,770 MM patients died. In 2014, the International Myeloma Working Group (IMWG) revised the diagnostic criteria for MM and divided the disease development of MM into three stages, namely, monoclonal immunoglobulinemia (MGUS), smoky myeloma (SMM) and active myeloma....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/118C12Q2600/156
Inventor 钱自亮陈红王白云徐文胜
Owner SUZHOU HONGYUAN BIOTECH CO LTD
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