Kit for detecting NPM (Nucleophosmin)1 gene mutation types

Abstract

The invention relates to kit for detecting NPM (Nucleophosmin)1 gene mutation types. The kit comprises an RT-PCR (Reverse Transcription-Polymerase Chain Reaction) solution, a primer and probe mixed solution, an RT-PCR reaction enzyme system, DEPC (Diethylpyrocarbonate) H20 and a packaging box which is used for separating and intensively packaging reagent bottles or tubes. Since the kit applies a one-step real-time fluorescence PCR (Polymerase Chain Reaction) mode and carries out LNA (Locked Nucleic Acid) modification on a specific probe, mutation of exons 12 of an NPM1 gene in a bone marrow and peripheral blood sample of an acute myeloid leukemia patient can be hypersensitively and rapidly detected, and the minimum detectable quantity is 0.2ng; meanwhile, A type mutation, B type mutation and D type mutation can be classified according to different channel signal values; meanwhile, the sample extracting effect can be detected by adding an internal standard control system, and the kit can be widely applied to estimation of an acute myeloid leukemia chemotherapeutic effect.

Description

technical field [0001] The invention relates to a hypersensitive typing kit for detecting NPM1 gene mutation nucleic acid, in particular to a method for simultaneously detecting NPM1 gene mutation A by using multiple real-time fluorescent polymerase chain reaction technology and locking nucleic acid modification probes in one tube reaction Type, B type, D type kits. Background technique [0002] Acute myelogenous leukemia (AML) is a malignant clonal blood disease originating from hematopoietic stem / progenitor cells. It is characterized by abnormal proliferation of primitive and immature myeloid cells in bone marrow and peripheral blood. Anemia, hemorrhage, infection and leukemic cell infiltration in organs are the main features. Epidemiological surveys show that AML is the most common adult acute leukemia, with an annual incidence of about 3 / 100,000. Nucleophosmin (NPM1) is one of the main protein molecules located in the nucleolar granule region, located on the long arm o...

AI Technical Summary

Problems solved by technology

Different alleles have different distributions of restriction enzyme sites, resulting in DNA fragment bands of different lengths. This method does not require low cost and easy interpretation, but the operation is cumbersome and the sensitivity is low.

RT-qPCR is also known as one-step reverse transcription fluorescent quantitative PCR method. It is a technology that achieves one-step completion of reverse transcription and fluorescent quantitative PCR through temperature adjustment. The operation process is carried out in a closed tube, which will not cause sample contamination. Wide range, but the detection site needs to be clear about the mutation type

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