Kit for detecting NPM (Nucleophosmin)1 gene mutation types

A kit and gene technology, applied in the field of ultra-sensitive typing kits, can solve the problems of high detection sensitivity, low sensitivity and complicated operation, and achieve the effects of improving detection efficiency, high throughput and simple operation.

Active Publication Date: 2016-09-21
DAAN GENE CO LTD
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  • Description
  • Claims
  • Application Information

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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 quantitativ

Method used

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  • Kit for detecting NPM (Nucleophosmin)1 gene mutation types
  • Kit for detecting NPM (Nucleophosmin)1 gene mutation types
  • Kit for detecting NPM (Nucleophosmin)1 gene mutation types

Examples

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Effect test

Embodiment 1

[0025] Example 1 Preparation and use of the kit for NPM1 gene mutation typing

[0026] 1. Prepare a kit including the following components

[0027] 1 tube of primer-probe mixture (25 μl / tube), 1 tube of RT-PCR reaction solution (125 μl / tube), 1 tube of RT-PCR enzyme system (75 μl / tube), 1 tube of positive quality control (200 μl / tube), Negative quality control (200μl / tube) 1 tube, DEPC H 2 O (2000μl / tube) 1 tube.

[0028] 2. Sample extraction

[0029] Extract in vitro transcribed RNA of NPM1 gene type A mutation, No. 1 (0.2ng), No. 2 (2ng), No. 3 (20ng), No. 4 (200ng), in vitro transcribed RNA of NPM1 gene type B mutation, No. 5 (0.2ng), No. 6 (2ng), No. 7 (20ng), No. 8 (200ng), in vitro transcription RNA of NPM1 gene D mutation, No. 9 (0.2ng), No. 10 (2ng), No. 11 ( 20ng), No. 12 (200ng), and 3 cases of clinical samples confirmed by Sanger sequencing method were extracted at the same time, No. 13 (A-type mutation), No. 14 (B-type mutation), No. 15 (D-type mutation), and t...

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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...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6886C12Q2600/118C12Q2600/156C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2563/107C12Q2521/107C12Q2545/101
Inventor 李明何皖平刘悦蒋析文高秀洁温楚茵张文苑邱秋淳
Owner DAAN GENE CO LTD
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