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Kit for rapid detection of common mutant genes of thalassemia

A technology for thalassemia and mutated genes, which is applied in the field of kits for rapid detection of common mutated genes of thalassemia, and can solve problems such as cumbersome operations, short amplification fragments, and contamination of PCR products

Active Publication Date: 2017-10-20
亚能生物技术(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the detection conditions and operation methods of these two methods are inconsistent, they must be operated separately, while PCR-RDB is cumbersome to operate, and the amplified fragment is short, which is prone to PCR product contamination
The detection range of the kits developed in the early stage also has certain limitations, so the adjustment of the detection range and the improvement of the detection method will provide support for the prevention and control of thalassemia

Method used

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  • Kit for rapid detection of common mutant genes of thalassemia
  • Kit for rapid detection of common mutant genes of thalassemia
  • Kit for rapid detection of common mutant genes of thalassemia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0103] Embodiment 1: The detection result of using the kit of the present invention in known genotype samples

[0104] 1. Composition of the kit

[0105] (1) Primers, including:

[0106] 1.1) Primers for detection of α-thalassemia mutation: design primers for detection of α-thalassaemia mutation according to the sequence of the α-globin gene cluster (NG_000006.1) published by NCBI, wherein -- SEA 、-- THAI , -α 2.4 , -α 21.9 Primers were designed by Gap-PCR method, -α 4.2 and-alpha 3.7 Then it is done based on the relative quantification of the Y1 segment and the Y2 segment, α CS α, α QS α, α Westmead α and Hb Q-Thailand used AS-PCR to design primers;

[0107] 1.2) Primers for detecting β-thalassaemia mutation: designed according to the sequence of β-globin gene cluster (NG_000007.3) published by NCBI, wherein HPFH-SEA and G gamma + ( A γδβ) 0 Primers were designed by Gap-PCR method, β -90 , β -29 , β -28 , β Cap+40-43 , β Int M , β Int CD , β CD14-15 , β ...

Embodiment approach

[0136] The instrument used for PCR reaction was Bio-Rad real-time thermal cycler CFX96. The PCR reaction program is as follows: the PCR reaction program is 95°C for 10 minutes, 94°C for 30 seconds, 62.5°C for 30 seconds and 72°C for 30 seconds, 28-35 cycles, and 62°C for 60 minutes to extend.

[0137] Sample processing: DNA was extracted with a universal DNA kit, diluted with double distilled water to 20-50 ng / μL for later use.

[0138] Sample detection: Configure the known genotype samples (20 copies, respectively numbered 1#, 2#...20#) according to the Tube1 system and Tube2 system, place them in the PCR machine and run the PCR reaction program. After the completion of the PCR program, take 0.4 μl of the PCR product of Tube1 detection tube and Tube2 detection tube of PCR product respectively, the internal molecular weight reference of the mixed sequencing system, and 10 μl of the mixed sequencing loading carrier deionized formamide (HiDi), and use the sequencer after mixing...

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Abstract

The invention discloses a kit for rapid detection of common mutant genes of thalassemia. The kit includes primers designed specific to --<SEA>, --<THAI>, -alpha<2.4>, -alpha<21.9>, HPFH-SEA, DBT, -alpha<4.2>, -alpha<3.7>, alpha<CS>alpha, alpha<QS>alpha, alpha<Westmead>alpha, Hb Q-Thailand, beta<-90>, beta<-29>, beta<-28>, beta<Cap+40-43>, beta<Int M>, beta<Int CD>, beta<CD14-15>, beta<CD17>, beta<CD26>, beta<27 / 28>, beta<IVS-I-1>, beta<IVS-I-5>, beta<CD37>, beta<CD41-42>, beta<CD43>, beta<CD71-72>, beta<CD95>, and beta<IVS-II-654> and gender detection primers, and the primers are subpackaged in 2 tubes. The kit provided by the invention can rapidly and accurately detect the common mutant genes of thalassemia.

Description

technical field [0001] The invention relates to a kit for detecting thalassemia, in particular to a kit for rapidly detecting common mutation genes of thalassemia. Background technique [0002] Thalassemia is a monogenic genetic disease with high incidence in tropical and subtropical regions. Patients with severe β-thalassemia require routine blood transfusion and iron deflection therapy, and their quality of life is poor. Guangxi and Guangdong are areas with a high incidence of thalassemia, and about 7% of the population are carriers of the β-thalassemia gene. Due to the high carrier rate of thalassemia-causing alleles in the population in this area, the probability of marriage among carriers of the same type of thalassemia alleles is also high, resulting in a higher probability of newborns with severe thalassemia. The way to reduce the birth probability of patients with severe thalassemia is to strengthen the screening of thalassemia among the married and childbearing po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6883C12Q2600/156C12Q2600/16
Inventor 黄德珍
Owner 亚能生物技术(深圳)有限公司
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