Primer and probe combination and kit for multiplex real-time fluorescence PCR detection of beta-thalassemia gene mutation

Abstract

The invention provides a primer and probe combination and a kit for multiplex real-time fluorescence PCR detection of beta-thalassemia gene mutation, and belongs to the technical field of molecular biology detection. The primer and probe combination comprises one or more of a first group of primer and probe combination to a twelfth group of primer and probe combination; each group of primer and probe combination comprises primers and probes for detecting two beta-thalassemia gene mutation types and two corresponding wild types; and conditions of the mutation types are indicated by an FAM fluorescence channel or an HEX fluorescence channel, and the wild types are indicated by an ROX fluorescence channel or a CY5 fluorescence channel. The primer and probe combination can simultaneously detect two or more sites in the mutation types and the wild types of a beta-thalassemia mutant gene, and reads information and a detection result through four different fluorescence channels, so that high-throughput and specific detection of beta-thalassemia is achieved.

Description

technical field [0001] The invention relates to the technical field of molecular biology detection, in particular to a combination of primers and probes and a kit for multiple real-time fluorescent PCR detection of beta-thalassemia gene mutation. Background technique [0002] β-thalassemia is a hemolytic anemia caused by the deletion or defect of the β-strain protein gene, which inhibits the synthesis of the β-strain protein chain. The main mutation method is point mutation, and a few are deletion mutations. Completely incapable of synthesizing β chains is called β0 thalassemia, can partially synthesize β chains and is called β+ thalassemia, which can be formed through different combinations: β0 thalassemia homozygote (β0 / β0), β0 thalassemia double heterozygote (β0 / β+ ), β0 heterozygous (β0 / βA), β+ thalassemia homozygous (β+ / β+) and β+ thalassemia heterozygous (β+ / βA). Among them, patients with β-thalassemia major are β+ thalassemia homozygotes, β0 thalassemia homozygotes, ...

AI Technical Summary

Problems solved by technology

At present, some studies choose to use real-time fluorescent PCR technology for the detection of β-thalassemia gene mutations, but they all detect one site in a single tube, and cannot detect multiple sites in a single tube, and cannot perform high-throughput detection

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