Method and kit for detecting alpha and beta thalassemia point mutation based on next generation sequencing technology

Abstract

The invention relates to a method and kit for detecting alpha and beta thalassemia point mutation based on a next generation sequencing technology .In the detection method, PCR (polymerase chain reaction) amplification primer sequences SEQ ID NO1-17 in exon, regulation and transcription regions of HBA1, HBA2 and HBB genes, PCR marker primer sequences SEQ ID NO18-115 at the 5'-terminal and PCR marker primer sequences SEQ ID NO116-213 at the 3'-terminal involved with alpha and beta thalassemia are included. The detection method comprises the following detection steps: (1) constructing a next generation sequencing library; (2) purifying the next generation sequencing library; (3) sequencing through a next generation sequencer; and (4) performing bioinformatic analysis to obtain a result. The kit for detection comprises the above-mentioned primer sequences and the next generation sequencer solexa. The detection method provided by the invention has the characteristics of simple operation steps, high detection specificity, wide detection range, low cost and the like.

Description

technical field [0001] The invention relates to a method and kit for detecting α and β thalassemia point mutations based on next-generation sequencing technology, belonging to the field of molecular biology. Background technique [0002] Thalassemia, or thalassemia, is a group of inherited hemolytic anemia diseases caused by partial or complete inhibition of globin peptide chain synthesis due to globin gene deletion or point mutation. The clinical manifestations vary according to the severity of the disease. Patients mainly present with anemia, jaundice, and hepatosplenomegaly. The globin peptide chains are divided into α, β, γ, and δ, and thalassemia mainly has four types: α, β, δβ, and δ, and the former two (α, β thalassemia) are the most common. The total carrier rate among the population in Guangxi, my country is as high as 24.5%. There is currently no effective treatment for this disease, and it is recognized as the preferred preventive measure both at home and abroad...

AI Technical Summary

Problems solved by technology

However, the current PCR-reverse dot hybridization method for molecular diagnosis of non-deletion thalassemia has a very limited detection range, including 17 kinds of β point mutations and 3 kinds of α point mutations (α WS 、α QS 、α CS ), it is very likely to cause missed diagnosis, resulting in the birth of a severely ill child

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