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Kit for childhood acute lymphoblastic leukemia genotyping

An acute lymphocyte and genotyping technology, applied in the field of biomedicine, can solve the problems of cumbersome operation steps, unstable reaction system, and low sensitivity, and achieve the effects of stable experimental system, improved sensitivity, and easy operation

Inactive Publication Date: 2017-08-11
BEIJING CHILDRENS HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has the advantages of low experimental cost and short detection period, but the operation steps are cumbersome, the sensitivity is low and the reaction system is unstable

Method used

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  • Kit for childhood acute lymphoblastic leukemia genotyping
  • Kit for childhood acute lymphoblastic leukemia genotyping
  • Kit for childhood acute lymphoblastic leukemia genotyping

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1, collection, acquisition and processing of clinical samples

[0034] 1. Collection of clinical specimens

[0035] Bone marrow samples from 219 newly diagnosed children with ALL who were treated in Beijing Children's Hospital between May 2011 and August 2016 were collected (with the consent of the children themselves or their guardians, and signed informed consent).

[0036] 2. Diagnosis and classification of newly diagnosed children with acute lymphoblastic leukemia

[0037] 219 children with ALL were diagnosed as ALL by MICM classification, including 16 children with T-ALL (T lineage ALL, T-ALL) and children with progenitor B lineage ALL (pro-B-ALL) 9 cases, 3 cases of pre-B-ALL (precursor Blineage ALL, pre-B-ALL) children, 191 cases of common B-ALL (common B lineage ALL, c-B-ALL) children. The diagnostic and typing criteria were based on the national unified standard (Suggestions for Diagnosis and Treatment of Children with Acute Lymphoblastic Leukemia....

Embodiment 2

[0067] Embodiment 2, GeXP multiple gene genetic analysis system detection

[0068] 1. Synthesis of primers

[0069] 57 typing genes, 3 internal reference genes (B2M, PSMC4 and GUSB) and 2 quality control genes (pcDNA3.1(+) and Kan R ) primer sequence has been disclosed in the Chinese invention patent document CN 104059970 B, and the specific information is shown in Table 3.

[0070] Table 3. Primer sequences and grouping of 57 typing genes, 3 internal reference genes and 2 quality control genes

[0071]

[0072]

[0073] Note: Sequence 1-126 is the primer sequence after adding the universal Tag sequence; the fragment size* refers to the size of the amplified fragment after removing the universal Tag sequence.

[0074] 2. GeXP reaction system optimization

[0075] 1. Preparation of mixed RNA samples

[0076] Randomly take RNA samples from 16 newly diagnosed ALL children numbered 1-16 in Table 4, and first use DNA / RNase-free H 2 O were diluted to 50ng / μL, and then mix...

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PUM

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Abstract

The invention discloses a kit for childhood acute lymphoblastic leukemia genotyping. The kit includes a primer pair group A, a primer pair group B, a primer pair group C, UDG enzyme and dUTP. Each primer pair group consists of 57 primer pairs specific to 56 typing genes, wherein the 57 primer pairs are divided into three groups, and each group correspond to a GeXP amplification reaction. An experiment proves that the kit can be used for an RT-PCR one-step method, so that operation is simpler and more convenient, the pollution possibility is reduced, and the sensitivity is improved. In addition, a dUTP / UDG technology is adopted to prevent pollution, so that an experimental system is more stable, the accuracy and specificity are greatly improved, and the kit has an important application value.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a kit for genotyping children with acute lymphoblastic leukemia. Background technique [0002] Leukemia is a malignant proliferative disease of the hematopoietic system, and it is one of the diseases that seriously threaten the life and health of children. Among them, acute lymphoblastic leukemia (ALL) is the most common type, accounting for 75%-80% of childhood leukemia. Currently, it is recognized that ALL in children is a heterogeneous tumor with changes in multiple genes and multiple signaling pathways. Accurate diagnosis and typing at the first visit is the basis for rational clinical treatment and good prognosis. [0003] At present, the international diagnosis and typing of children's ALL are the classification of cell morphology (Morphology), immunology (Immunology), cytogenetics (Cytogenetics) and molecular biology (Molecular biology), that is, MICM typing. According to the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6886C12Q2600/112C12Q2600/158C12Q2600/16C12Q2531/113C12Q2537/143C12Q2565/125
Inventor 郑胡镛李杰吴勇孙嫣然张瑞东高超陆继冉张巧生陈真许丽亭李静
Owner BEIJING CHILDRENS HOSPITAL AFFILIATED TO CAPITAL MEDICAL UNIV