Test kit for detecting thalassemia-related gene mutation and application thereof

A technology of thalassemia and kits, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., and can solve the problems of low sensitivity and single biological inspection

Active Publication Date: 2015-02-25
BOAO BIOLOGICAL CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the current biological detection is single and the sensitivity is not high, so it is of practical significance to provide a primer set and kit for detecting thalassemia

Method used

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  • Test kit for detecting thalassemia-related gene mutation and application thereof
  • Test kit for detecting thalassemia-related gene mutation and application thereof
  • Test kit for detecting thalassemia-related gene mutation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Amplification and detection of thalassemia gene by primer set

[0047] The components of PCR amplification system 1, 2, 3, and 5 are the same: the total volume is 25 μL, and the composition is: solvent 1×buffer, dNTP 0.1~0.2mM, MgCl 2 1~3mM, hot start Taq DNA polymerase 1-1.5U.

[0048] Four sets of primers corresponding to the PCR amplification system:

[0049] Primer set 1: a primer pair for identifying nt29A>G (as shown in SEQ ID No.1-3), a primer pair for identifying Cap(-ACCC) (as shown in SEQ ID No.7-9), A pair of primers for identifying CD14-15 (+G) (as shown in SEQ ID No.13-15), a pair of primers for identifying CD26G>A (as shown in SEQ ID No.19-21), and a pair of primers for identifying CD26G>A (as shown in SEQ ID No.19-21). The primer pair for identifying IVS-I-1G>T (as shown in SEQ ID No.25-27), the primer pair for identifying CD41-42 (-TCTT) (as shown in SEQ ID No.31-33), The concentration of each upstream primer is 0.1 μM, and the concentration...

Embodiment 2

[0058] Example 2 Detection of the thalassemia gene by the kit provided by the present invention

[0059] The components of PCR amplification systems 1, 2, 3, and 5 are the same: the total volume is 25 μL, and the composition is: solvent 1×buffer, dNTP 0.1-0.2mM, MgCl21-3mM, hot-start Taq DNA polymerase 1-1.5U.

[0060] Four sets of primers corresponding to the PCR amplification system:

[0061] Primer set 1: a primer pair for identifying nt29A>G (as shown in SEQ ID No.1-3), a primer pair for identifying Cap(-ACCC) (as shown in SEQ ID No.7-9), A pair of primers for identifying CD14-15 (+G) (as shown in SEQ ID No.13-15), a pair of primers for identifying CD26G>A (as shown in SEQ ID No.19-21), and a pair of primers for identifying CD26G>A (as shown in SEQ ID No.19-21). The primer pair for identifying IVS-I-1G>T (as shown in SEQ ID No.25-27), the primer pair for identifying CD41-42 (-TCTT) (as shown in SEQ ID No.31-33), The concentration of each upstream primer is 0.1 μM, and t...

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Abstract

The invention relates to the biotechnology field, in particular to a primer group and application thereof, and a test kit and application thereof. The primer group used for amplifying thalassemia gene nucleotide fragments includes a primer group A and / or a primer group B. Primers amplify gene fragments in which group-related gene loci are located. The primer group and the test kit can detect wild type results and mutant type results of alpha and beta thalassemia effectively.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer set and its application, a kit and its application. Background technique [0002] Thalassemia, formerly known as thalassemia, is a group of inherited hemolytic anemia diseases. Anemia or pathological condition caused by the absence or insufficient synthesis of one or more globin chains in hemoglobin due to inherited genetic defects. Due to the complexity and diversity of gene defects, the type and quantity of globin chains lacking and the variability of clinical symptoms are large. [0003] The molecular structure and synthesis of globin chains are determined by genes. The gamma, delta, epsilon, and beta globin genes make up the "beta gene family," and the zeta and alpha globin genes make up the "alpha gene family." Normal people inherit two α-globin genes (αα / αα) from both parents to synthesize enough α-globin chains; inherit one β-globin gene from both parents to synthe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6883C12Q2600/156
Inventor 李阳项光新邢婉丽
Owner BOAO BIOLOGICAL CO LTD
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