Gene probe composition and kit for acute lymphocytic leukemia detection

A technology of acute lymphocytes and gene probes, applied in the direction of recombinant DNA technology, microbial measurement/inspection, biochemical equipment and methods, etc., can solve problems such as deletion of heterozygosity

Active Publication Date: 2013-03-20
INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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Problems solved by technology

The TEL gene is frequently deleted in childhood acute lymphoblastic leukemia and, alth

Method used

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  • Gene probe composition and kit for acute lymphocytic leukemia detection
  • Gene probe composition and kit for acute lymphocytic leukemia detection
  • Gene probe composition and kit for acute lymphocytic leukemia detection

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Experimental program
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Embodiment 1

[0072] The present invention will be further described below through specific examples, but protection scope of the present invention is not limited. Embodiment 1: The preparation method of IKZF1 gene probe comprises the following steps:

[0073] 1. Primer design and clone screening: The IKZF1 gene is located in the 12.3-12.2 segment of the short arm of human chromosome 7 (7p12.3-p12.2), and all clones containing the IKZF1 gene were searched in the UCSC genome browser, NCBI Clone Registry, and Ensembl Genome Browser databases , and the optimal clone containing the gene was screened by polymerase chain reaction, numbered RP11-663L2 (as shown in Table 2).

[0074]2. Cloning culture and identification: purchase clone RP11-663L2 (Invitrogen, USA), take 8 microliters of clone bacteria solution and add it to 5 milliliters of LB liquid medium containing chloramphenicol resistance, shake and activate for 12 hours at 37°C ; Then add all the bacterial liquid to 450 ml of LB liquid medi...

Embodiment 2

[0101] Embodiment 2: the preparation method of TEL, AML1, PAX5, P16, IKZF1 gene probe composition, comprises the following steps

[0102] 1. Primer design and clone screening:

[0103] By searching UCSC genome browser, NCBI Clone Registry, Ensembl Genome Browser and other databases, all clones containing TEL, AML1, PAX5, P16, and IKZF1 genes were screened out by polymerase chain reaction, and the numbers were: RP11 -654E9, RP11-77G18, RP11-243F8, RP11-97A22, RP11-663L2. (As shown in Table 1).

[0104] 2. Cloning culture and identification: Purchase clones (Invitrogen, USA) according to the clone number, take 8 microliters of clone bacteria liquid and add them to 5 milliliters of chloramphenicol-resistant LB liquid medium, shake and activate for 12 hours at 37°C Then all the bacterial liquid was added to 450 milliliters of LB liquid medium containing chloramphenicol resistance, and the bacterial liquid was collected after shaking and culturing at 37°C for 12 hours.

[0105] ...

Embodiment 3

[0133] The results of fluorescence in situ hybridization showed that in the same nucleus, each probe obtained two clear fluorescent signals, and 5 probes obtained a total of 10 corresponding fluorescent signal points. Green-TEL (green), PF555-AML1 (red), PF590-PAX5 (orange), HyPer5-P16 (purple), PF415-IKZF1 (blue) multicolor images obtained from ( image 3 ), it can be seen that the fluorescence signals of the five different genes detected at the same time are all strong, the images are clear, and the signal-to-noise ratio is high. The method is reliable and stable. Embodiment 3: A fluorescent in situ hybridization detection kit for acute lymphoblastic leukemia (50 parts), the composition is as follows:

[0134] 1) Fluorescence-labeled probe set hybridization mixture 100 microliters; 1 tube

[0135] 2) DAPI counterstain solution 500 microliters; 1 tube

[0136] 3) 20ml of 20X SSPE washing solution; 1 bottle

[0137] 4) 1 instruction manual.

[0138] The concentration of t...

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Abstract

The present invention relates to a gene probe composition and a kit for acute lymphocytic leukemia detection. The gene probe composition comprises a TEL gene probe, an AML1 gene probe, a PAX5 gene probe, a P16 gene probe and an IKZF1 gene probe. The present invention further provides an acute lymphocytic leukemia fluorescence in situ hybridization detection kit, wherein the kit comprises the gene probe composition. With the kit, concurrent five gene detection can be performed on the same sample even the single leukemia cell, and detection capability and efficiency are significantly improved.

Description

technical field [0001] The invention relates to a gene probe composition and a kit, in particular to a gene probe composition and a kit for detecting acute lymphocytic leukemia. Background technique [0002] Acute lymphoblastic leukemia is a highly malignant blood disorder characterized by large numbers of immature white blood cells similar to lymphoblasts in the peripheral blood, these abnormal cells can also be found in the bone marrow, lymph nodes, spleen and other organs Find. Acute lymphoblastic leukemia accounts for about 80% of acute leukemia in children, and the peak incidence is between 3 and 7 years old. The disease can also occur in adults, accounting for about 20% of all adult leukemia. In acute leukemia, malignant cells lose their ability to mature and differentiate, and malignant cells rapidly divide and replace normal cells. Bone marrow failure occurs when malignant cells replace normal bone marrow components, and the patient becomes prone to bleeding and in...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11G01N21/64
Inventor 程涛竺晓凡胡林萍张丽缪为民
Owner INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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