ALL prognostic related gene mutation detection method, primers and kit

A detection method and gene technology, applied in the fields of medicine and molecular biology, can solve the problems of insufficient accuracy and convenience, and achieve the effect of less experimental equipment, simple operation and high accuracy

Inactive Publication Date: 2016-11-23
北京海思特医学检验实验室有限公司
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Problems solved by technology

[0005] Aiming at the problems of insufficient accuracy and convenience in the current ALL prognosis assessment detection technology, the present invention provides a new ALL prognosis-related gene mutation detection method and corresponding primers and kits

Method used

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  • ALL prognostic related gene mutation detection method, primers and kit
  • ALL prognostic related gene mutation detection method, primers and kit
  • ALL prognostic related gene mutation detection method, primers and kit

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Experimental program
Comparison scheme
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Embodiment approach

[0079] The specific implementation is as follows:

[0080] In step S1, the blood of the ALL patient diagnosed with clinical specimens is collected, and the genomic DNA is extracted; the genomic DNA can be extracted using a conventional kit column extraction method, and operated according to the instructions of the kit used, and the concentration of 50 μL collected is 50-100 ng / μL DNA solution, which can be directly used for detection or stored at -20°C;

[0081] In step S2, a solution is prepared using the specific primers in the kit, wherein the concentration of the specific primers is 5 pmol / μL; each pair of the specific primers includes an upstream primer and a downstream primer for matching the corresponding Specific segments of the exons of the CREBBP gene containing hotspot mutations were amplified by PCR. The primer sequences and the exons corresponding to the amplification are shown in Table 1 below:

[0082]

[0083]

[0084] Under the premise of meeting the d...

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Abstract

The embodiment of the invention provides an ALL prognostic related gene mutation detection method, primers and a kit. PCR and sequencing technologies are used, specific primers are mainly used for amplifying specific exon sections of CREBBP genes in genome DNA, sequencing is conducted on target fragments obtained through amplification through a sequencing reaction, and whether mutation exists or not is detected by comparing the obtained sequence with a standard genetic sequence. The detection process is rapid and sensitive, operation is easy, the accuracy is high, and the detection result is definite and clear; the detection specificity is high, a gene mutation detection method which is the most accurate at present is adopted, and purposiveness is high; the blood collection amount of a detection specimen is low, a detected object himself/herself is benefited, fewer experimental instruments are used in the detection process, and the cost is reduced. The invention aims at obtaining analytical data capable of being used for medication guidance and prognosis evaluation of acute lymphoblastic leukemia, and the analytical data can play an important role in the field of medical detection.

Description

technical field [0001] The invention belongs to the technical field of medicine and molecular biology, and in particular relates to a method for detecting mutations of genes related to prognosis of ALL (Acute Lymphoblastic Leukemia, acute lymphoblastic leukemia). The invention also provides primers used in the detection method and a kit containing the primers. Background technique [0002] ALL is a neoplastic disease originating from B-lineage or T-lineage lymphoid progenitor cells. The blast cells proliferate and aggregate abnormally in the bone marrow and inhibit normal hematopoiesis, resulting in anemia, thrombocytopenia and neutropenia; blast cells can also invade Extramedullary tissues, such as meninges, gonads, thymus, liver, spleen, or lymph nodes, cause corresponding lesions. In 2008, the WHO diagnostic criteria for ALL were: cell morphology, immunophenotype, bone marrow primitive and immature lymphocytes ≥ 20%, classified as: ① precursor B-cell acute lymphoblastic ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q1/6869C12Q2600/118C12Q2600/156C12Q2531/113
Inventor 陈欣傅廷娇张娟李悠南程宇王绪华梁超陈忠黄士昂
Owner 北京海思特医学检验实验室有限公司
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