A kit for rapid detection of common β-thalassaemia mutant alleles in Chinese population

An allele and kit technology, applied in the field of on-site gene kits, can solve the problems of short amplified fragments, cumbersome operations, and PCR product contamination, etc.

Active Publication Date: 2020-04-14
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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  • A kit for rapid detection of common β-thalassaemia mutant alleles in Chinese population
  • A kit for rapid detection of common β-thalassaemia mutant alleles in Chinese population
  • A kit for rapid detection of common β-thalassaemia mutant alleles in Chinese population

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Experimental program
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Effect test

Embodiment 1

[0071] Embodiment 1: The detection result of using the kit of the present invention in known genotype samples

[0072] 1. Composition of the kit

[0073] (1) Primer

[0074] According to the β-globin gene cluster (NG_000007.3) sequence published by NCBI, 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 CD17 , β CD26 , β 27 / 28 , β IVS-I-1 , β IVS-I-5 , β CD37 , β CD41-42 , β CD43 , β CD71–72 , β CD95 and beta IVS-II-654 Primers were designed by AS-PCR method, where β -28 , β CD17 , β CD26 , β 27 / 28 , β IVS-I-1 , β CD41-42 , β CD43 , β CD71–72 , β IVS-II-654 Normal control SNP detection primers and AMXY primers for gender detection were designed. The specific primer names and sequences are shown in Table 4 below:

[0075] Table 4:

[0076]

[0077]

[0078]In Table 4, "M" represents a mutation site, "N" represents a normal control site, FAM rep...

Embodiment approach

[0090] Place the PCR reaction system with template DNA on a general-purpose PCR instrument. 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 seconds. minute extension.

[0091] Sample processing: DNA was extracted with a universal DNA kit, diluted with double distilled water to 20-50 ng / μL for later use.

[0092] Sample detection: Run the PCR program in the PCR reaction system of the known genotype to be tested. After the PCR program runs, take 0.4 μl of PCR product. 10 μL, after mixing, the sequencer was used for loading and sequencing, and the fragment analysis method was used to detect the results.

[0093] 3. Sample source: All samples are derived from DNA samples whose genotypes were determined by conventional Gap-PCR technology or sequencing technology.

[0094] 4. Data analysis and result determination: The GeneMapper software was used to read and analyze the results, and...

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PUM

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Abstract

The invention discloses a kit for quickly detecting beta Mediterranean anemia mutant alleles common in Chinese population. The kit comprises primers designed for deletion-type beta Mediterranean anemia genes HPFH-SEA and G gamma+(A gamma delta gamma)0, primers designed for non-deletion-type Mediterranean anemia alleles 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 AMXY primer for detecting gender. The kit can quickly and accurately detect the beta Mediterranean anemia mutant alleles common in the Chinese population.

Description

technical field [0001] The invention relates to a kit for detecting thalassemia, in particular to a kit for rapidly detecting the common β-thalassemia mutation allele in Chinese population. 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 m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/6858
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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