Primer set and kit for detecting rare deletion type thalassemia

A thalassemia and primer set technology, applied in biochemical equipment and methods, recombinant DNA technology, microbe measurement/inspection, etc., can solve the problems of undetected rare thalassemia, and achieve convenient detection, simple operation, and cost saving Effect

Active Publication Date: 2014-02-26
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

[0015] (1) At present, the kits for the diagnosis of deletional α-thalassemia clinically used in my country are all developed based on the principle of multiple Gap-PCR, such as Yaneng Biotechnology (Shenzhen) Co., Ltd., Shenzhen Yishengtang Biotechnology Co., Ltd. and Guangzhou Daan The α-thalassemia detection kits developed by gene companies can only realize the three common missing thalassemia genotypes (-- SEA , -α 3.7 , -α 4.2 ) detection
[0016] (2) At present, the kits for the diagn

Method used

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  • Primer set and kit for detecting rare deletion type thalassemia
  • Primer set and kit for detecting rare deletion type thalassemia
  • Primer set and kit for detecting rare deletion type thalassemia

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Experimental program
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Effect test

Embodiment 1

[0075] 1. Primer design and screening:

[0076] The present invention utilizes various missing primers reported in the literature to confirm collected clinically known genotype positive samples, and uses the positive samples as positive control templates to perform primer screening and PCR program optimization.

[0077]Considering that there can be as many as 5-6 pairs of primers in the multiplex PCR system, and the high homology and high GC content among the α gene clusters, it is impossible to achieve good conditions for simultaneous amplification with the β gene, so the α, β Two types of primer amplification split the operation. Based on the positions of various rare deletion breakpoints reported in the literature (see Table 2) and the sequences in the related gene database, we designed Gap-PCR-specific amplification primers at both ends to apply multiplex PCR to multiple genes. The purpose is to detect the same tube type (the two tubes correspond to α and β thalassemia re...

Embodiment 2

[0121] Use of the kit for detecting rare deletion thalassemia with PCR reaction solution A and PCR reaction solution B in Example 1:

[0122] 1. Intended use

[0123] For 11 known rare deletion types in China (-- 11.1 、--FIL 、-- THAI , -α 27.6 , -α 2.4 , -α 2.8 Equal α-thalassemia deletion genotype and β Chinese(Aγδβ)0 , β SEAβ0 , β Thai(Aγδβ)0 , β Yunnanese(Aγδβ)0 , β Taiwaneseβ0 and other β-thalassemia deletion genotypes) to achieve qualitative detection of the above-mentioned 11 known rare deletion types, providing a comprehensive and reliable basis for genetic screening of thalassemia. When doing phenotypic and genetic analysis, for patients with intermediate or severe α-thalassemia phenotype, use the existing α-thalassemia detection kit (for -α 3.7 , -α 4.2 、-- SEA ) detects a 1.8kb electrophoresis band (suggesting that the result is normal genotype αα / αα), or detecting only a 1.3kb band (suggesting that the result is SEA homozygote -- SEA / -- SEA ) can dete...

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Abstract

The invention belongs to the technical field of biology and particularly relates to a primer set and a kit for detecting rare deletion type thalassemia. The primer set and the kit can be directly used for rapidly and stably detecting domestic 11 types of known rare deletion type thalassemia. The primer set for detecting the rare deletion type thalassemia comprises a primer set A and a primer set B, wherein the primer set A comprises 12 primers for detecting alpha-thalassemia deletion gene types; the primer set B comprises 10 primers for detecting beta-thalassemia deletion gene types. The kit can adopt a multiplex Gap-PCR (Polymerase Chain Reaction) technology to detect 11 rare deletion gene types and can be used for directly detecting 11 rare deletion thalassemia gene types for one time. The kit is simple to operate and saves time and cost when being compared with a manner of combining a nested PCR with a genetic analysis in antenatal diagnosis; conditions are created for comprehensively carrying out thalassemia screening and scientific evidences are provided for thalassemia diagnosis of premarital detection, antenatal detection and fetuses of the pregnancy.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a primer set and a kit for detecting rare deletion thalassemia. Background technique [0002] Thalassemia is one of the most common human monogenic blood diseases in the world. It is listed by the World Health Organization as six common diseases that endanger human health. It is also the most common and most harmful genetic disease in southern China. [0003] α-Thalassemia (hereinafter referred to as α-thalassemia) is a group of genetic hemolytic hemoglobinopathies characterized by reduced or non-synthesized α-globin chains and an imbalance in the ratio of α-chain / non-α-chain. The most common type of deletion in my country is -α 3.7 , -α 4.2 、-- SEA 3 types, with the further elucidation of the mechanism of α-thalassemia, some new gene deletion types have been discovered one after another, for example- 11.1 、-- FIL 、-- THAI , -α 27.6 , -α 2.4 , -α 2.8 ,-- 11.1 、-...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q2537/143
Inventor 李印淑刘福平刘晶晶任维
Owner 亚能生物技术(深圳)有限公司
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