Fluorescence detection kit for detecting deafness susceptibility gene GJB2 235delC and application of fluorescence detection kit

Abstract

The invention discloses a fluorescence detection kit for detecting deafness susceptibility gene GJB2 235delC. The kit comprises a pre-amplification reagent and a post-amplification reagent; the pre-amplification reagent comprises a reaction mixture comprising a PCR (Polymerase Chain Reaction) buffer solution, MgCl2 and dNTPs (deoxyribonucleotide Triphosphates), a Taq enzyme, ultrapure water and a primer mixture comprising high specificity amplification GJB2 235delC and an Amelogenin gene locus; and the post-amplification reagent comprises a genotyping standard substance and an interior label. According to the invention, a fluorescence labeling technology, a LNA (Locked Nucleic Acid) nucleoside monomer doping and primer modification technology and a capillary electrophoresis technology are firstly compounded to be adopted; the simultaneous detection on the high sensitivity specificity of the deafness gene locus GJB2 235delC is implemented; manpower and material resources and time are greatly saved; and the pollution caused by multi-step operation is prevented.

Description

technical field [0001] The invention relates to a fluorescent detection kit for detecting deaf disease susceptibility gene GJB2 235delC, which belongs to the technical field of biological detection. Background technique [0002] Worldwide studies on the pathogenic factors of hearing and speech disabilities show that about 60-80% of the patients are caused by genetic factors, and clinical research data from developed countries show that hereditary deafness accounts for about 80% of deafness patients. Therefore, in the past ten years, the research on the pathogenesis and molecular epidemiology of hereditary deafness has become one of the most important contents of deaf research. With the completion of the Human Genome Project, great progress has been made in the positioning and cloning of deafness genes. The molecular genetics research and molecular epidemiological data of deafness have enabled researchers to gradually realize that mutations in susceptibility genes for deafnes...

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Problems solved by technology

These methods have different defects, such as cumbersome operation, difficult interpretation of results, poor repeatability, many false negatives and false positives, etc.

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