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High-throughput assay method for leukaemia driver genes

A detection method and gene-driven technology, which can be used in biochemical equipment and methods, microbial determination/inspection, etc.

Pending Publication Date: 2018-02-23
JIAXING YUNYING MEDICAL INSPECTION CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, usually the clinical progress of leukemia is very fast, and this detection method may often lead to wrong selection and delay the diagnosis of the disease.

Method used

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  • High-throughput assay method for leukaemia driver genes
  • High-throughput assay method for leukaemia driver genes
  • High-throughput assay method for leukaemia driver genes

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0044] A kind of detection method of leukemia driver gene of the present invention, comprises the steps:

[0045] 1. Sample library preparation:

[0046] (1) Ultrasound fragmentation: the initial amount is 3ug, add nuclease-free water to 100ul and dilute to 30ng / ul. SCIENTZ08-Ⅲ cup-type ultrasonic cell pulverizer was used for ultrasonic fragmentation, and the setting parameters were: power 70%, interruption for 3s, stop for 1s, and cycle for 30-60min.

[0047] (2) Take out the magnetic beads half an hour in advance to return to room temperature, shake and mix, take 180ul of magnetic beads and add them to the PCR product interrupted by ultrasound, beat and mix, and incubate in the greenhouse for 5 minutes.

[0048] (3) Put the PCR single tube on the magnetic stand, let it stand for 5 minutes, remove the supernatant, keep the PCR tube on the magnetic stand, add 200ul of 80% ethanol (newly prepared) to rinse, let stand for 30s, remove the supernatant, 80 Rinse twice with % etha...

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Abstract

The invention discloses a high-throughput assay method for leukaemia driver genes. By choosing mutant genes and fusion genes commonly seen in leukaemia at present, in combination with the second-generation high-throughput sequencing technology, the method can more comprehensively assay the gene mutation and fusion conditions of leukaemia. Compared with the classical Sanger sequencing method mostlyadopted at present, the second-generation high-throughput sequencing technology has the advantages of high throughput, short sequencing time, high precision, low cost and rich information, and can accurately locate and analyze target gents in a short time. With the help of the high-throughput assay technology, the method can provide more detailed gene mutation and fusion spectra for clinical doctors to comprehensively judge the prognosis of leukaemia and guide individualized treatment.

Description

technical field [0001] The invention relates to a high-throughput detection method for leukemia driving genes, in particular to a detection method for leukemia gene mutations. Background technique [0002] Leukemia, also known as "blood cancer", is a malignant clonal disorder of hematopoietic stem cells. People of almost all ages are at risk of developing leukemia. In 2012, about 352,000 people worldwide suffered from various types of leukemia, and 265,000 of them died of this malignant tumor. About 90 percent of all leukemia patients are adults. Leukemia ranks first in the incidence of pediatric malignant tumors, and its mortality rate ranks first in malignant tumors that cause death in children and adults under the age of 35. [0003] There are many types of leukemia, which are divided into acute and chronic according to the speed of disease progression, mainly including four types: Chronic Myelocytic Leukemia (CML), Chronic Lymphocytic Leukemia (CLL), Acute Myelogenous...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6869
CPCC12Q1/6869C12Q2535/122C12Q2565/519C12Q2531/113
Inventor 张道允巩子英叶建伟王伟
Owner JIAXING YUNYING MEDICAL INSPECTION CO LTD
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