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Rapid detection kit aiming at thalassemia mutant genes common in Chinese crowds

A technology for detecting kits and mutant genes, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve the problems of short amplified fragments, cumbersome operations, and contamination of PCR products

Active Publication Date: 2017-10-13
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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  • Rapid detection kit aiming at thalassemia mutant genes common in Chinese crowds
  • Rapid detection kit aiming at thalassemia mutant genes common in Chinese crowds
  • Rapid detection kit aiming at thalassemia mutant genes common in Chinese crowds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1: Using the kit of the present invention to detect results in samples with known genotypes

[0093] 1. The composition of the kit

[0094] (1) Primer, including:

[0095] 1.1) Primer for detection of α-thalassaemia mutation: According to the sequence of α-globin gene cluster (NG_000006.1) published by NCBI, the detection primer for α-thalassaemia mutation is designed, among which - SEA , - THAI , -Α 2.4 , -Α 21.9 Design primers using Gap-PCR method, -α 4.2 And -α 3.7 According to the relative quantification of Y1 zone and Y2 zone, α CS α, α QS α and α Westmead α Design primers using AS-PCR method;

[0096] 1.2) Primer for detection of β-thalassaemia mutation: The detection of β-thalassaemia is designed according to the sequence of β-globin gene cluster (NG_000007.3) published by NCBI, among which HPFH-SEA and G γ + ( A γδβ) 0 (DBT) Design primers by Gap-PCR method, β -90 , Β -29 , Β -28 , Β Cap+40-43 , Β Int M , Β Int CD , Β CD14-15 , Β CD17 , Β CD26 , Β 27 / 28...

Embodiment approach

[0117] The instrument used for the PCR reaction is the Bio-Rad real-time thermal cycler CFX96. 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 extension.

[0118] Sample processing: DNA is extracted by the universal DNA kit and diluted with double distilled water to 20-50ng / μL for later use.

[0119] Sample detection: add 20 samples of known genotypes to be tested (numbered 1#, 2#...20#) into the PCR reaction system, run the PCR program, and take 0.4μl PCR after the PCR program runs The product, the molecular weight internal reference of the mixed sequencing system, liz500, and the mixed sequencing loading carrier deionized formamide (HiDi) 10μl, were mixed with a sequencer for loading and sequencing, and the results were detected by the fragment analysis method.

[0120] 3. Sample source: All samples are derived from DNA samples whose genotype has been determined by conventio...

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Abstract

The invention discloses a rapid detection kit aiming at thalassemia mutant genes common in Chinese crowds. The rapid detection kit comprises primers designed aiming at <SEA>, <THAI>, alpha<2.4>, alpha<21.9>, HPFH SEA and DBT, primers designed aiming at alpha<4.2> and alpha<3.7> and primers designed aiming at alpha<CS>alpha, alpha<QS>alpha, alpha<Westmead>alpha, 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>. The kit can quickly and accurately detect the thalassemia mutant genes common in Chinese crowds.

Description

Technical field [0001] The invention relates to a kit for detecting thalassemia, in particular to a rapid detection kit for common thalassemia mutant genes in Chinese population. Background technique [0002] Thalassemia is a single-gene genetic disease that occurs frequently in tropical and subtropical regions. Patients with severe β-thalassemia require routine blood transfusion and iron removal treatment, and their quality of life is poor. Guangxi and Guangdong are areas with high incidence of thalassemia, and about 7% of the population are carriers of the β-thalassemia gene. Due to the high rate of carrying alleles of thalassaemia in the population in this area, the probability of marriage of carriers of the alleles of homotype thalassaemia is also higher, resulting in a higher probability of birth of severe thalassaemia. The way to reduce the birth rate of patients with severe thalassaemia is to strengthen the screening of thalassaemia among married and child-bearing people...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6883C12Q2600/156C12Q2600/16C12Q2531/113C12Q2537/143C12Q2563/107
Inventor 黄德珍
Owner 亚能生物技术(深圳)有限公司
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