Relative quantification method and kit for detecting copy number of human DMD gene by multiple real-time fluorescence PCR

Abstract

The invention discloses a relative quantification method and kit for detecting the copy number of a human DMD gene through multiple real-time fluorescence PCR. The kit comprises an exon reaction solution I, an exon reaction solution II, an exon reaction solution III, a main reaction mixed solution, a positive reference substance, a negative reference substance and a blank reference substance. According to the relative quantification method and kit for detecting the copy number of the human DMD gene through the multiple real-time fluorescence PCR, the 4th, 17th and 45th exons, 8th, 50th and 52nd exons, 47th, 48th and 51st exons of the DMD gene are respectively amplified through three independent PCR reactions, and each of three reaction solutions comprises four fluorescence channels, so that the quantitative detection of the copy number of the 4th, 8th, 17th, 45th, 47th, 48th, 50th, 51st and 52nd exons of the DMD gene is realized.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a relative quantitative method and kit for detecting the copy number of human DMD gene by multiple real-time fluorescent PCR method. Background technique [0002] Duchenne / Beckman muscμLar dystrophy (DMD / BMD) is an X-linked recessive genetic disease characterized by progressive muscle weakness and muscle atrophy caused by DMD gene mutations leading to dystrophin protein deficiency Hereditary Muscle Disease. Its clinical features include: muscle cramps, myalgia, quadriceps myopathy, asymptomatic hypercreatine kinaseemia, X-linked dilated cardiomyopathy, etc. [0003] The clinical manifestations of female DMD gene defect carriers are quite different. The severe cases can be typical DMD manifestations, and the mild cases can be manifested as mild proximal muscle weakness, gastrocnemius pseudohypertrophy, or basically normal muscle function. The incidence of DMD in mal...

AI Technical Summary

Problems solved by technology

[0014] Although the multiplex fluorescent quantitative PCR technology has the above-mentioned advantages, the 427m transcript of the DMD gene has 79 exons. If the fluorescent quantitative detection is performed on all 79 exons, it will not be possible to operate, and all the above-mentioned advantages will be gone. no deposit

Although the DMD gene has 79 exons, the deletion or duplication of exons is not randomly distributed, but there are mutation hotspots

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