Joint detection method of fusion genes related to leukemia and diagnosis kit

A diagnostic kit and gene fusion technology, applied in biochemical equipment and methods, microbial determination/inspection, measuring devices, etc., can solve problems such as poor repeatability, cumbersome operation, and insufficient sensitivity, and achieve rapid and stable detection Good performance and high sensitivity

Active Publication Date: 2011-01-26
MEDI GENETECH
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

Immunological methods have high false positive and false negative rates, and cannot achieve early diagnosis
Among the molecular biology methods widely used at home and abroad to detect leukemia fusion genes, fluorescence in situ hybridization (FISH) can only perform qualitative detection, and the operation is compl

Method used

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  • Joint detection method of fusion genes related to leukemia and diagnosis kit
  • Joint detection method of fusion genes related to leukemia and diagnosis kit
  • Joint detection method of fusion genes related to leukemia and diagnosis kit

Examples

Experimental program
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Example Embodiment

[0065] Example 1: Liquid chip combined parallel detection method for multiple fusion genes related to 3 kinds of leukemia

[0066] The specific detection method includes the following steps:

[0067] 1. Preparation of microsphere mixture for detecting SIL-TAL1 (type II, type III), MLL-AF4 (e9 / e5, e9 / e4), CBFB-MYH11 (typeD) fusion genes

[0068] 1. Synthesize oligonucleotide probes according to the following sequence:

[0069] SIL-TAL1 (type II, type III): 5'-AminolinkerC12 AGTTACGCTGCGGTGTGGTC-3', as shown in SEQ ID NO.1;

[0070] MLL-AF4 (e9 / e5, e9 / e4): 5'-AminolinkerC12 TATTGCTGTCAAAGGAGGCGG-3', as shown in SEQID NO.2.

[0071] CBFB-MYH11 (type D): 5'-AminolinkerC12 TGTCCTTCTCCGAGCCTCTTCA-3', as shown in SEQ ID NO.3;

[0072] Abelson gene: 5'-AminolinkerC12 CTGAAGGGCTTCTTCCAGAT-3', as shown in SEQ ID NO.12;

[0073] 2. Coupling the above oligonucleotide probes containing amino modification with 4 kinds of carboxyl microspheres numbered 11, 15, 17, and 21 respectively

[0074] 2.1 Take out...

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Abstract

The invention discloses a joint detection method of fusion genes related to leukemia and a diagnosis kit. The method comprises the following steps of: designing a primer and a probe through a fusion gene mRNA (the fusion genes related to the leukemia are SIL-TAL1(type II, type III), MLL-AF4(e9/e5,e9/e4) and CBFB-MYH11(type D)) related to the leukemia; hybridizing a probe and microsphere mixture formed by the covalent bonding of the probe and microspheres with an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) amplification product; and adding phycoerythrin as streptavidin biotin to detect fluorescent signals of different microspheres so as to confirm whether a sample to be detected contains a leukemia fusion gene as well as the expression conditions of the fusion gene. The method and the kit of the invention have the advantages of high sensitivity and flux, rapid and accurate detection, and the like and can be used for simultaneously, qualitatively and quantitatively detecting various leukemia fusion genes.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnosis and detection, in particular to a liquid phase chip joint detection method and a diagnostic kit for multiple fusion genes related to leukemia. Background technique [0002] Leukemia is a malignant tumor of the hematopoietic system caused by abnormal differentiation and malignant proliferation of hematopoietic stem cells. It is one of the most common malignant tumors in my country. With the continuous progress of leukemia research, it has been found that many leukemia patients have specific chromosomal translocations, leading to the generation of new fusion genes, and these abnormal genes have become molecular biology-specific markers of different types of leukemia. In 2000, WHO's criteria for leukemia classification included some common abnormal genes as the criteria for the basic diagnosis of leukemia. Therefore, the detection of leukemia-related fusion genes at the molecular gene lev...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N21/64
CPCG01N33/57426C12Q2600/158C12Q1/6886
Inventor 邵棠徐春雷
Owner MEDI GENETECH
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