Kit for combined detection of acute myeloid leukemia

A combined detection technology for acute myeloid cells, applied in the field of molecular biological detection, can solve problems such as increased detection time and complex data analysis process

Active Publication Date: 2020-12-18
苏州科贝生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, its data analysis process is complicated, which increases the detection time

Method used

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  • Kit for combined detection of acute myeloid leukemia
  • Kit for combined detection of acute myeloid leukemia
  • Kit for combined detection of acute myeloid leukemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Example 1 A kit for combined detection of acute myeloid leukemia

[0099] Included in this kit:

[0100] (1) The special linker for DNA library construction includes 8 dimers composed of linker primer 1 and 8 different i5-terminal primers.

[0101] The sequences of the linker primer 1 and 8 different i5-terminal primers are shown in Table 4.

[0102] Table 4 Sequence list of special linker primers

[0103] Primer name Primer sequence 5_index sequence SEQ ID NO Adapter Primer 1 GATCGGAAGAGCCACATACTGA - SEQ ID NO:1 Primer 1 at i5 end AATGATACGGCGACCACCGAGATCTACACCCTCTCTATACACTCTTTCCCTACACGACGCTCTTCCGATCT CTCT CTAT SEQ ID NO:2 Primer 2 at i5 end AATGATACGGCGACCACCGAGATCTACACTATCCCTCTACACTCTTTCCCTACACGACGCTCTTCCGATCT TATC CTCT SEQ ID NO:3 Primer 3 at the i5 end AATGATACGGCGACCACCGAGATCTACACGTAAGGAGACACTCTTTCCCTACACGACGCTCTTCCGATCT GTAA GGAG SEQ ID NO:4 Primer 4 at the i5 end AATGATACGGCGACCACCGAGATCTACACACTG...

Embodiment 2

[0126] Example 2 A method of using a kit for joint detection of acute myeloid leukemia

[0127] The kit of this embodiment is the kit provided in Example 1, Figure 7 For the experimental flow chart of the detection method, Figure 8 Flowchart for single primer amplification library construction. The detection method of the kit comprises the following specific steps:

[0128] 1. Reverse transcription of cDNA

[0129] 1. First strand cDNA synthesis

[0130] 1.1 Reagent preparation: thaw the extracted sample RNA slowly on ice, mix it with a pipette, mix the random primer first-strand synthetase mixture, and the first synthesis buffer (purchased from Tiangen Biochemical Technology Co., Ltd., product type TIANSeq cDNA synthesis module, catalog number NG308-T1) Take out from -20 ℃, flick and mix well after thawing.

[0131] 1.2 Establish the following reaction system in the PCR tube, and mix well by gently blowing with a pipette:

[0132] RNAXμL (1000ng);

[0133] Random prime...

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Abstract

The invention provides a kit for combined detection of acute myeloid leukemia. The kit comprises a special linker for DNA library construction, a specific composite primer for detecting gene rearrangement and gene whole exon mutation, and a composite primer for library amplification. The special linker is 8 dimers formed by a linker primer 1 and eight different i5-terminal primers respectively; the types of the gene rearrangement and the gene whole exon mutation are CBFB rearrangement, RUNX1 rearrangement, MLL rearrangement, RARA rearrangement, MECOM rearrangement and TP53 gene whole exon mutation; and the composite primer for library amplification has different i7 terminals. The kit provided by the invention can be used for detecting a plurality of common molecular genetic variations of acute myeloid leukemia at one time, and is simple to operate, short in time and high in detection efficiency.

Description

technical field [0001] The invention belongs to the field of molecular biological detection, in particular to a kit for combined detection of acute myeloid leukemia. Background technique [0002] Acute myeloid leukemia (AML) is a malignant disease of myeloid hematopoietic stem cells. It is mainly characterized by abnormal proliferation of primitive and immature myeloid cells in bone marrow and peripheral blood. The clinical manifestations are anemia, hemorrhage, infection and fever, visceral infiltration, abnormal metabolism, etc. Most cases are acute and severe, and the prognosis is dire. [0003] In recent years, through the study of gene mutation frequency and some susceptibility biomarkers, it has been found that acute myeloid leukemia may be the result of a combination of genetics and environmental factors. Acute myeloid leukemia (AML) is a heterogeneous disease comprising many entities with distinct genetic abnormalities and clinical features. [0004] Studying the g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6886C12Q2600/156C12Q2600/16C12Q2525/191C12Q2525/173C12Q2531/113C12Q2535/122C12Q2537/143
Inventor 孙石磊姬云
Owner 苏州科贝生物技术有限公司
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