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Gene chip for leukemia diagnosis and treatment

A gene chip and leukemia technology, applied in the field of molecular medicine, can solve the problems of inability to early diagnosis, risk assessment and individualized treatment of children, low overall control level, and expensive detection costs, so as to shorten hospitalization time, reduce medical expenses, and improve diagnosis. The effect of rate and efficacy

Inactive Publication Date: 2012-09-19
CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, due to the various methods such as cell morphology detection, flow cytometry, nested RT-PCR, and conventional chromosome banding techniques currently used for patient typing and prognosis judgment, the required experimental conditions and technical personnel training requirements The cost of detection is high, which limits the large-scale implementation of MICM typing and diagnosis in primary hospitals, so that most of the leukemia patients hospitalized in primary hospitals in my country are still limited to morphological detection, and cannot be completely and accurately diagnosed. Early diagnosis, risk assessment and individualized treatment with low overall control

Method used

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  • Gene chip for leukemia diagnosis and treatment
  • Gene chip for leukemia diagnosis and treatment
  • Gene chip for leukemia diagnosis and treatment

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The preparation of embodiment 1 gene chip

[0024] 1. Probe Design and Synthesis

[0025] The aforementioned chip is mainly divided into a gene expression detection unit and a gene polymorphism (SNP) detection unit, wherein the gene expression detection unit includes 18 fusion gene oligonucleotide probes and 48 expression gene oligonucleotide probes ; SNP unit contains gene SNP 47 kinds of probes.

[0026] The fusion gene oligonucleotide probe of the gene expression detection unit of the kit can specifically detect the fusion gene, which provides an important basis for leukemia diagnosis, typing, clinical treatment and prognosis judgment, and is also the basis for detection of leukemia minimal residual disease. Objects include BCR / ABL a2b3, BCR / ABL a2e1, BCR / ABLa 2b2, TEL / ABL, TEL / PDGFR, E2A / PBX1, ETV6-RUNX1, TEL-AML1, CBFβ-MYH11, MLL-AFX, MLL-AF1p, Any one or more of MLL-AF10, MLL-AF4, MLL-AF9, AML-ETO, PML / RARa, NPM / RAR and SET / CAN.

[0027] The expressed gene olig...

Embodiment 2

[0034] Embodiment 2 Chip performance testing

[0035] 1. Gene chip hybridization

[0036] Hybridization experiments to determine specificity were performed in duplicate, while other microarray hybridization experiments were performed in triplicate. 15 μl of standard hybridization solution contained 3×sodium citrate buffer solution (SSC), 1 μg of unlabeled herring egg DNA (Promega, Madison, USA), 0.3% sodium dodecyl sulfate (SDS) and 5 μl of the target PCR product. The volume of the hybridization solution used for testing the sensitivity and detecting the target gene is 2 μl, and the concentration of the components contained therein is the same as before. The hybridization solution is directly added to the exact position where the fixed DNA array is, and then covered with a cover slip. The fluorescein-labeled DNA was diluted with hybridization solution, denatured at 100°C for 2 minutes, cooled to ambient temperature, first added to the coverslip, and then placed on the covers...

Embodiment 3

[0040] Application of Embodiment 3 Chip

[0041] The detection process of this gene chip is the same as that of other gene chips containing gene oligonucleotide probes.

[0042] 188 cases of leukemia, including 35 cases of acute myeloid leukemia (AML) and 153 cases of acute lymphoblastic leukemia (ALL), were tested, total RNA was extracted, and then the RNA was reverse transcribed into cDNA, and the detection results were analyzed using this chip , see the table below.

[0043]

[0044]

[0045]

[0046]

[0047] Compared with traditional methods of morphology, immunology, cytogenetics and molecular biology, the results are as follows:

[0048]

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Abstract

The invention relates to the field of molecular medicine, and in particular relates to a gene chip for leukemia diagnosis and treatment. The chip contains oligonucleotide probes with sequences in SEQ ID NO:1-137. The chip has the following beneficial effects that: the actual demands in the blood system tumor diagnosis and treatment process in China are met; the gene chip is customized by screening key indicators in examination indicators in leukemia morphology, immunology, cytogenetics and molecular biology; and the multifunctional gene chip integrates early diagnosis, early warning, prevention and diagnosis and treatment of leukemia and can improve the individual diagnosis efficiency and treatment effect of leukemia, shorten the length of stay, reduce the medical expenses and improve the technical levels of the primary-level medical and health institutions.

Description

technical field [0001] The invention relates to the field of molecular medicine, in particular to a gene chip for diagnosis and treatment of leukemia. Background technique [0002] The incidence of leukemia has been increasing year by year, and it has become the number one killer of adolescents. The goal of treatment for childhood leukemia is no longer simply to pursue the remission rate, but to use comprehensive laboratory test results and a prognosis evaluation system to treat the disease according to risk factors. Children are grouped so that each child can receive individualized treatment of appropriate intensity and improve their long-term quality of life. In the past 20 years, the standardized treatment based on morphology, immunology, cytogenetics, molecular biology (MICM) and risk factors and the prevention of central nervous system leukemia have become the key to the success of childhood leukemia treatment. Minimize the adverse reactions and long-term complications...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C40B40/06
Inventor 邹琳谭俊杰张鹏辉
Owner CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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