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Primer and probe for detecting relative fusion genes of leukemia and kit of primer and probe

A fusion gene, leukemia technology, applied in biochemical equipment and methods, microbial determination/test, DNA/RNA fragments, etc., can solve the problems of difficult clinical application, complicated operation, poor accuracy, etc.

Active Publication Date: 2014-03-19
上海源奇生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have defects such as poor accuracy and complicated operation, making it difficult to apply them clinically.

Method used

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  • Primer and probe for detecting relative fusion genes of leukemia and kit of primer and probe
  • Primer and probe for detecting relative fusion genes of leukemia and kit of primer and probe
  • Primer and probe for detecting relative fusion genes of leukemia and kit of primer and probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0120] Table 15:

[0121]

[0122] in:

[0123] The RT-PCR reaction solution 1 for detecting the BCR-ABL fusion gene contains: 5 μl of 5×Buffer, 0.25 μl of 10 mmol / L dNTPs, 0.12 μl of 50 μmol / L of SEQ ID NO: 1 to SEQ ID NO: 5, 50μmol / L SEQ ID NO: 50.1μl, make up to 8μl with process water;

[0124] Table 16:

[0125] name

sequence

modify

SEQ ID NO: 1

CTGGCCCAACGATGGCGA

none

SEQ ID NO: 2

TCCGCTGACCATCAATAAGGA

none

SEQ ID NO: 3

TGGAGGAGGTGGGCATCTAC

none

SEQ ID NO: 4

CACTCAGACCCTGAGGCTCAA

none

[0126] SEQ ID NO: 5

CCTTCAGCGGCCAGTAGCATCTGA

5' Fam, 3' Tamara

[0127] The RT-PCR reaction solution 2 for detecting the PML-RARa fusion gene contains: 5 μl of 5×Buffer, 0.25 μl of 10 mmol / L dNTPs, 0.12 μl of 50 μmol / L of SEQ ID NO: 6 to SEQ ID NO: 10, 50μmol / L SEQ ID NO: 10 0.1μl, make up to 8μl with process water;

[0128] Table 17:

[0129] name

sequence ...

Embodiment 2

[0155] Embodiment 2: Using the kit of Embodiment 1 to detect the sample.

[0156] 1. Sample processing: RNA nucleic acid was extracted with RNA extraction kit for samples 1-10; the OD of the extracted RNA was 260 / OD 280 Between 1.8 and 2.0. It is recommended to test the extracted RNA immediately, otherwise it should be stored below -80°C; add the extracted RNA to 200ul RNeas-Free water, mix and dissolve, and use this treatment solution as a template for related PCR reactions;

[0157] 2. Preparation of amplification reagents:

[0158] Take out tubes 1-7 from the kit, melt at room temperature and shake to mix, then briefly centrifuge for 1-8 seconds;

[0159] Preparation of sample PCR master mix system:

[0160] Take 18ul of RT-PCR reaction solution, add 2ul of mixed enzyme solution; prepare 4 tubes for a single sample;

[0161] Take 2 8ul of RT-PCR reaction solution, add 2ul of mixed enzyme solution; prepare 4 tubes for a single sample;

[0162] Take 38ul of RT-PCR reac...

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Abstract

The invention relates to detection of genotypes, in particular to a primer and a probe for detecting relative fusion genes of leukemia and a kit of the primer and the probe. The kit can qualitatively detect quantity of BCR-ABL, AML1-ETO and PML-RARa of the relative fusion genes of the leukemia in human marrow specimens so as to instruct clinical diagnosis and pharmacy of the leukemia.

Description

technical field [0001] The invention relates to the detection of genotypes, in particular to a primer and a probe for detecting leukemia-related fusion genes, and a kit thereof. Background technique [0002] Leukemia is a malignant tumor of the hematopoietic system, which is characterized by the malignant proliferation of one or more blood cells in the hematopoietic system such as bone marrow and lymph nodes, and infiltrates various tissues and organs in the body, resulting in the inhibition of normal hematopoietic tissue cells and the production of various diseases. symptom. Clinically, leukemia is often characterized by fever, hemorrhage, anemia, and enlarged liver, spleen, and lymph nodes. Leukemia can generally be divided into acute leukemia (AL) and chronic leukemia (CL) according to the natural course of the disease and the degree of cell development. According to the morphology and biochemical characteristics of leukemia cells, it can be divided into acute lymphoblas...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q2600/156C12Q1/6886C12Q2600/158
Inventor 熊慧程新建谢立群陈嘉铮包文静童光铨陈伟
Owner 上海源奇生物医药科技有限公司
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