Primer pairs and probes for detecting thalassemia genes, kit and use method

Abstract

The invention discloses primer pairs and probes for detecting thalassemia genes. The primer pairs and probes include primer pairs and probes of beta-thalassemia CD41 / 42 and alpha-thalassemia-SEA; theprimer pair of the beta-thalassemia CD41 / 42 is a sequence 1 and a sequence 2, and the probes are a sequence 3 and a sequence 4; and the primer pair of thealpha-thalassemia-SEA is a sequence 5 and a sequence 6 or a sequence 5 and a sequence 7, and the probes are a sequence 8 and a sequence 9. The invention further provides a use method of the primer pairs and probes for detecting the thalassemia genes. The use method comprises the following steps of S1, collecting a sample and extracting DNA; S2, carrying out an amplification reaction by utilizing one kind of primer pair and probes; S3, detecting the fluorescence intensity and determining the genotype; and S4, repeating the process by utilizing the other kind of primer pair and probes. The method has the beneficial effects that the fluorescence detection accuracy is enhanced through the designed primer pairs and probes and the two kinds of primer pairs of the same type; and rapid, accurate and harmless detection is realized by extracting free DNA of plasma and blastocyst culture media of pregnant women.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to primer pairs, probes, kits and usage methods for detecting thalassemia genes. Background technique [0002] Thalassemia is a genetic hemolytic disease caused by globin synthesis disorders, including two types of α-thalassemia and β-thalassemia. It is more common in the south of my country, especially in Guangdong, Guangxi and Hainan. It is the single gene disease with the highest incidence and the greatest impact in the provinces south of the Yangtze River. We focused our study on the most common mutation, a mutation in the β-globin gene in β-thalassemia, a deletion of four nucleotides (–CTTT) at codons 41 / 42, HBB:c.126delcttt( CD41 / 42). CD41 / 42 mutations have the highest incidence among all β-thalassemia mutations in Guangxi, Guangdong, Hainan, and Sichuan, China, accounting for 36.4%, 42.5%, 73.7%, and 33.33% of all β-thalassemia alleles, respectively. The SEA-deletio...

AI Technical Summary

Problems solved by technology

ddPCR is now more commonly used in the detection of genetically modified foods, but it has not been used in the prenatal diagnosis of single-gene diseases.

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