Single-tube amplification kit for simultaneously detecting alpha and beta thalassemia genes

A thalassemia and kit technology, applied in the genetic field, can solve problems such as time-consuming, high operating requirements, and products of α-thalassemia and β-thalassemia gene amplification have not yet appeared

Active Publication Date: 2014-12-17
GUANGDONG HYBRIBIO BIOTECH CO LTD +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These products have high operating requirements, cumbersome steps, and long time-consuming, which cannot meet the simple and fast requirements of clinical diagnosis.
[0010] So far, there is no product that can be used to amplify α-thalassemia and β-thalassemia genes in the same reaction tube in clinical use for detection of thalassemia.

Method used

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  • Single-tube amplification kit for simultaneously detecting alpha and beta thalassemia genes
  • Single-tube amplification kit for simultaneously detecting alpha and beta thalassemia genes
  • Single-tube amplification kit for simultaneously detecting alpha and beta thalassemia genes

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0055] 1. Design of specific primers for amplifying the DNA in the sample to be tested and determination of the PCR reaction system

[0056] 1-1 Primer design

[0057] According to the sequence data of α-thalassemia gene deletion, α-thalassemia gene mutation and β-thalassemia gene mutation, 9 primers were designed for PCR amplification, and the 9 primer combinations were placed in the same reaction tube for multiplex PCR. The α-thalassemia and β-thalassemia genes were simultaneously amplified.

[0058]

[0059] 1-2 Determination of Multiplex PCR Reaction System

[0060] Through a large number of experimental comparisons, the control of Mg 2＋ Concentration, primer concentration and PCR Enhancer concentration can achieve simultaneous high-efficiency amplification of a-thalassemia gene and β-thalassemia gene with good specificity. The final optimal PCR reaction system is shown in Table 1: 45 μl of PCR reaction solution, 5 μl of DNA sample, and a total reaction volume of 50 μl...

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Abstract

The invention discloses a single-tube amplification kit for simultaneously detecting alpha and beta thalassemia genes, and the kit comprises (1) a gene chip on which a probe is arranged as SEQID No:1-37 and its complementary sequence; (2) one group of primers which is used for multiple PCR (polymerase chain reaction) and regarded as SEDID No. 38-46. Through the kit, alpha and beta thalassemia genes can be simultaneously amplified in the same reaction tube, and three deleted alpha-thalassemia genes (SEA, -alpha 3.7 and -alpha 4.2), three mutant alpha-thalassemia genes (CS, QS and WS) and 19 mutant beta-thalassemia genes can be simultaneously detected. In the same reaction tube, alpha-thalassemia genes and beta-thalassemia genes can be also simultaneously amplified, thereby greatly improving the diagnostic efficiency and accuracy, reducing the cost and shortening he detection time, thus the kit has the great significance of thalassemia crowd screening, genetic counseling and antenatal diagnosis.

Description

technical field [0001] The invention relates to the field of genes, in particular to a single-tube amplification and simultaneous detection of α and β thalassemia gene kits. Background technique [0002] Thalassemia (referred to as thalassemia) is a hemolytic anemia caused by an imbalance in the synthesis rate of α, β-globin peptide chains due to human gene mutation or deletion. Thalassemia is one of the most common genetic diseases in the world, and it is widely distributed in Mediterranean countries and other areas where malaria used to be high. In my country, the provinces south of the Yangtze River have a high incidence rate. In addition to Guangdong, Guangxi, and Hainan provinces with the highest incidence rates, Guizhou, Yunnan, and Sichuan are also high-incidence areas. The two common types of thalassemia are alpha-thalassemia and beta-thalassemia. [0003] α-thalassemia is a chronic hemolytic disease caused by autosomal genetic defect, which causes the loss or dysf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6827C12Q1/6858C12Q2531/113C12Q2537/143C12Q2565/501
Inventor 苏彬峰李烈军
Owner GUANGDONG HYBRIBIO BIOTECH CO LTD
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