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Detection kit for combined detection of chronic myeloid leukemia

A technology for combined detection of chronic myeloid cells is applied in the field of combined detection of chronic myeloid leukemia detection kits, which can solve the problems of increased detection time and complex data analysis process.

Active Publication Date: 2021-01-26
苏州科贝生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

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

Method used

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  • Detection kit for combined detection of chronic myeloid leukemia
  • Detection kit for combined detection of chronic myeloid leukemia
  • Detection kit for combined detection of chronic myeloid leukemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0095] Example 1 A kit for combined detection of chronic myelogenous leukemia

[0096] Included in this kit:

[0097] (1) Special linker for DNA library construction: the special linker is 8 dimers composed of linker primer 1 and 8 different i5-terminal primers.

[0098] The sequences of the linker primer 1 and 8 different i5-terminal primers are shown in Table 4.

[0099] Table 4 Special Adapter Primer List

[0100] 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

[0101] Table 4 continued

[0102] Pri...

Embodiment 2

[0129]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:

[0130] 1. Reverse transcription of cDNA

[0131] 1. First strand cDNA synthesis

[0132] 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.

[0133] 1.2 Establish the following reaction system in the PCR tube, and mix well by gently blowing with a pipette:

[0134] RNAXμL (1000ng);

[0135] Random primer 1μL;

[0136] Ultrapure water (10-X) μL;

[0137] Total volume 11 μL.

[0138] 1.3 I...

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Abstract

The invention provides a kit for combined detection of chronic myeloid leukemia. The kit comprises a special linker for DNA library construction, a specific composite primer for detecting gene mutation, fusion and deletion, and a composite primer for library amplification. The special linker is eight dimers formed by a linker primer 1 and eight different i5-terminal primers respectively; the typesof gene mutation, fusion and deletion are BCR / ABL fusion, FGFR1 rearrangement, JAK2 gene V617F mutation, JAK2 rearrangement, NUP98 rearrangement, PDGFRA rearrangement and PDGFRB rearrangement; and the library amplification composite primer has different i7 ends, and the sequence of the primer is SEQ ID NO:39-46. The kit can be used for detecting a plurality of common molecular genetic variationsof chronic 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 biology detection, and in particular relates to a detection kit for combined detection of chronic myeloid leukemia. Background technique [0002] Chronic myeloid leukemia (CML) is a malignant disease based on pluripotent stem cells, characterized by acquired abnormal chromosomes, and the main feature is the unlimited proliferation of granulocytes in bone marrow and peripheral blood. The impact of chronic myelogenous leukemia on the blood and bone marrow is mainly manifested in the production of a large number of immature white blood cells, which accumulate in the bone marrow and inhibit the normal hematopoiesis of the bone marrow; and can spread throughout the body through the blood, resulting in anemia, easy bleeding, and infection in patients. and organ infiltration. [0003] Specific cytogenetic changes occur in approximately 95% of CML patients and manifest as translocations of chromosomes 9 and 22 l...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6869C12N15/11
CPCC12Q1/6869C12Q1/6886C12Q2600/156C12Q2600/16C12Q2525/191C12Q2531/113C12Q2537/143C12Q2535/122
Inventor 孙石磊姬云
Owner 苏州科贝生物技术有限公司
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