Combined methylmalonic acidemia gene mutation detection kit

A technology of methylmalonic acidemia and detection kits, which is applied to the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., to achieve the effects of poor long-term prognosis, short time period and high precision

Active Publication Date: 2022-01-21
北京华诺奥美医学检验实验室有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no quick and easy detection kit for gene mutations in hotspot mutation regions of the gene (referring to gene segments where various major, new or sporadic mutations mainly occur)

Method used

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  • Combined methylmalonic acidemia gene mutation detection kit
  • Combined methylmalonic acidemia gene mutation detection kit
  • Combined methylmalonic acidemia gene mutation detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0025] The present invention establishes a primer and a kit for detecting the hotspot mutation region of human MMAHCC gene by using multiplex PCR-sanger sequencing technology, which is simple, fast, accurate and high-throughput, so as to provide a reference for the detection of MMAHC gene mutation. The kit is stored in -20°C. Kit specifications: 50 servings / box, see Table 1 for specific components.

[0026]

[0027]

[0028]

[0029]

[0030]

[0031]

[0032]

[0033]

[0034] Kit Performance Verification

[0035] 1. Sample processing

[0036] Enterprise reference products were selected for kit performance verification. All samples were subjected to nucleic acid extraction kit (Product No.: IVD4173) from Biogene, and the obtained DNA samples were stored at 2-8°C; if the samples were not used for a long time, they could be stored at -20°C.

[0037] 2. PCR amplification

[0038] Configure the PCR amplification reaction solution as shown in the table ...

experiment example

[0074] 60 clinical samples were tested.

[0075] 1.1 Sample processing

[0076] Blood samples were collected from 20 combined MMA subjects and 40 healthy subjects.

[0077] Key points for sample collection: Use purple cranial tubes (blood collection tubes containing ethylenediaminetetraacetic acid and its salts) to collect venous blood, and store it at 4°C for later use.

[0078] Take 200 µL from each sample, and extract DNA according to the DNA extraction kit (product number: IVD4173) of Biogene Biotechnology.

[0079] 1.2 PCR amplification

[0080] Configure the PCR amplification reaction solution as shown in the table below (45 μL for each reaction)

[0081] .

[0082] Aliquot the configured PCR amplification reaction solution into 45 μL of each reaction space. Add 5 μL each of the processed sample DNA, positive control DNA, and negative control DNA to the corresponding reaction wells, and perform PCR amplification on the machine.

[0083] 1.3 Amplification procedur...

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PUM

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Abstract

The invention belongs to the technical field of biological detection, and discloses a combined methylmalonic acidemia gene mutation detection kit. The kit is used for rapidly, accurately and sensitively detecting MMACHC gene mutation by designing three groups of primer pairs and a reaction hole aiming at MMACHC gene hotspot mutation regions. The application of the kit fills the blank of clinical MMACHC gene mutation detection, and experimental results are good in repeatability and high in precision. According to the invention, multiple mutation sites (including new mutation / non-found mutation) can be simultaneously detected at one time, the detection period is short, the detection can be completed within 8 hours at the soonest, the detection time is greatly saved, and the clinical efficiency is improved. Detection is completed through a multiple PCR and sanger sequencing technology, the requirement for operators is simple, the reagent cost is low, and clinical popularization is facilitated.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a detection kit for combined methylmalonic acidemia gene mutation. Background technique [0002] Methylmalonic acidemia (MMA) is a common organic acidemia, which is an autosomal recessive genetic disease. The clinical manifestations of the disease are recurrent vomiting, difficulty in feeding, lethargy, mental and physical development retardation or regression, convulsions, dyspnea and abnormal muscle tone, and yellow hair as the main manifestations. Acylcarnitine in blood tandem mass spectrometry and urinary methylmalonate detected by gas chromatography-mass spectrometry are the main diagnostic basis for methylmalonic acidemia. According to the level of homocysteine, it is divided into two types: simple methylmalonic acidemia and methylmalonic acidemia combined with homocysteinemia. The clinical manifestations of the disease vary greatly from individual to individ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/686C12Q1/6869C12N15/11
CPCC12Q1/6883C12Q1/686C12Q1/6869C12Q2600/156C12Q2537/143C12Q2535/101
Inventor 陈倩于超计肖江山郑萍张平平谢丽娜高彦彦毛莹莹冯硕姬辛娜魏星
Owner 北京华诺奥美医学检验实验室有限公司
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