Primer, probe and reaction buffer solution combinations used for multiplex real-time fluorescence PCR detection of alpha-thalassemia, and kit

Abstract

The invention provides primer, probe and reaction buffer solution combinations used for the multiplex real-time fluorescence PCR detection of alpha-thalassemia, and a kit, and belongs to the technicalfield of molecular biology detection. The combinations include eight sets of primer and probe combinations and four reaction buffer solutions; and the four reaction buffer solutions include a B-4 reaction buffer solution, a B-2 reaction buffer solution, J-1 reaction buffer solution and H-1 reaction buffer solution. Through the combinations, the amplification of --<SEA>, --<THAI>, -alpha<3.7> and-alpha<4.2> long fragment deletion and the non-deletion type synchronous typing detection amplification of alpha<CS>alpha / alpha alpha, alpha<QS>alpha / alpha alpha and alpha<WS>alpha / alpha alpha can berealized; and the method has the characteristics of being strong in specificity, high in sensitivity and short in time consumption, and can distinguish different mutation types through different fluorescence channels in a same reaction tube.

Description

technical field [0001] The invention relates to the technical field of molecular biology detection, in particular to primers, probes, reaction buffer combinations and kits for multiple real-time fluorescent PCR detection of α-thalassemia. Background technique [0002] α-thalassemia is caused by a defect in the α-globin gene located at the end of human chromosome 16. It can be divided into two types: deletion type and non-deletion type. Among them, the incidence of deletion type is much higher than that of non-deletion type. α-thalassemia is an autosomal single-gene genetic disease, which occurs frequently in southern China, especially in South China and Southwest China, and the Chinese population is mainly right-sided deletion type (-α 3.7 ), missing on the left (-α 4.2 ), and the Southeast Asian type (-- SEA ) three deletion types, α CS α / αα, α QS α / αα, α WS α / αα three non-deletion types and sporadic cases Thailand deletion type (-- THAI ) and other α-thalassemia geno...

AI Technical Summary

Problems solved by technology

Then through agarose gel electrophoresis, the genotype of the sample is detected according to the size of the electrophoresis fragment, which is an effective method for the detection of deletion thalassemia, but this method can only detect the deletion type in α-thalassemia

[0006] RDB technology is based on PCR amplification, hybridizes PCR products with specific probes immobilized on membrane strips, and then observes the results by exciting fluorescence or a series of color reactions. This method has high sensitivity, but this method can only Non-deletion thalassemia detected

[0007] There is currently a lack of a way to achieve -- SEA 、-- THAI , -α 3.7 , -α 4.2 Amplification of long deletions, and and α CS α / αα, α QS α / αα, α WS α / αα three non-deletion synchronous typing detection method

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