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Beta-thalassemia gene amplification kit and method based on recombinase polymerase amplification technology

A recombinase polymerase, thalassemia technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of PCR amplification dependence and slow amplification rate, and achieve high sensitivity and fast amplification time. Effect

Inactive Publication Date: 2018-10-09
GUANGDONG WOMEN & CHILDREN HOSPITAL
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  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, PCR amplification still has disadvantages such as relying on large instruments and slow amplification rate.

Method used

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  • Beta-thalassemia gene amplification kit and method based on recombinase polymerase amplification technology
  • Beta-thalassemia gene amplification kit and method based on recombinase polymerase amplification technology
  • Beta-thalassemia gene amplification kit and method based on recombinase polymerase amplification technology

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] (1) Primer design and optimization

[0056] Using bioinformatics knowledge and related bioinformatics software, analyze the β-globin gene sequence, design specific RPA amplification primers for RPA amplification, and analyze their sequences, especially the primer pair After the sequence analysis of the combination, the qualified primers were selected for experimental verification and optimization. Alternative primers are listed in Table 4.

[0057] Table 4 alternative PCR primer sequences (both 5'-3')

[0058]

[0059]

[0060] F in the table indicates the upstream primer, R indicates the downstream primer; bio indicates the modification of Biotin at the 5' end.

[0061] (1) Prepare each primer to 10 μM and set aside. Using human genomic DNA (extracted from peripheral blood, negative for thalassemia, Guangdong) as a template, the primers are optimized according to the following process: the present invention divides the β-globin gene into three fragments for am...

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Abstract

The invention discloses a beta-thalassemia gene amplification kit and a method based on a recombinase polymerase amplification technology. The kit comprises RPA amplification primers which are respectively P1F-1 shown as SEQ ID NO. 1, P1Rbio-1 shown as SEQ ID NO. 2, P2F-1 shown as SEQ ID NO. 3, P2Rbio-1 shown as SEQ ID NO. 4, P3F-1 shown as SEQ ID NO. 5 and P3Rbio-1 shown as SEQ ID NO. 6. The amplification is performed on genomic DNA of a to-be-detected specimen by using the RPA amplification primers and can be completed within 30 minutes, and the sensitivity is high; an obtained amplificationproduct can be detected by a liquid phase chip, and the accuracy rate is high.

Description

technical field [0001] The invention relates to a beta-thalassemia gene amplification kit and method, in particular to a beta-thalassemia gene amplification kit and method based on recombinase polymerase amplification technology. Background technique [0002] Thalassemia, referred to as thalassemia, is a genetic hemolytic disease caused by globin synthesis disorders, divided into α-thalassemia (abbreviated as α-thalassemia) and β-thalassemia (β-thalassemia) two types. The disease is mainly found in the countries along the Mediterranean Sea and Southeast Asian countries. It is the single gene disease with the widest distribution and the largest cumulative population in the world. There are about 90 million symptoms and gene carriers in the world. It is more common in the south of China, and it is one of the genetic diseases with the highest incidence and the greatest impact in the provinces south of the Yangtze River in my country, especially in Guangxi, Guangdong and Hainan...

Claims

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Application Information

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IPC IPC(8): C12Q1/6844
CPCC12Q1/6844C12Q2521/507C12Q2522/101C12Q2537/1376C12Q2565/501
Inventor 骆明勇胡听听胡思琪杨笑函张畅斌詹文丽王继成
Owner GUANGDONG WOMEN & CHILDREN HOSPITAL
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