Kit for detecting two pathogenic mutation loci in TAB2 gene of human congenital heart disease, and PCR (Polymerase Chain Reaction) amplification method thereof

Abstract

The invention discloses a kit for detecting two pathogenic mutation loci in a TAB2 gene of a human congenital heart disease, and a PCR (Polymerase Chain Reaction) amplification method thereof. The method is characterized in that a mutation detection primer modified by an anti-3'-5' excision enzyme in an enzyme digestion manner is applied into the PCR in which a high-fidelity DNA (Deoxyribose Nucleic Acid) polymerase is participated, so that the PCR specificity is improved; thus, the genetype of the corresponding mutation loci of the TAB2 gene can be judged by directly observing whether a gel electrophoresis product stripe exists or not. The kit has a low requirement to instruments and equipment, is simple to operate and economic and has no need of sequence measuring. As a result, the kit can be operated in an ordinary molecular biology laboratory.

Description

technical field [0001] The invention belongs to the clinical detection technology in the field of biomedicine, and relates to a detection method for human congenital heart disease gene pathogenic mutation sites. [0002] The invention provides a PCR kit for detecting the mutation of human congenital heart disease gene. The content of the present invention relates to a kit for detecting two disease-causing mutation sites in the TAB2 gene of congenital heart disease and a PCR amplification method thereof, and the detection result can be used for clinical auxiliary diagnosis of congenital heart disease. Background technique [0003] Congenital Heart Disease is the most common type of congenital malformation in children, which seriously endangers children's health and life. The incidence rate varies with the number of observed cases, about 0.4%-5%, of which 60% die in <1 year old. [0004] The types of congenital heart disease are generally considered to be simple ventricul...

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Problems solved by technology

[0010] In 2010, scientist Bernard Thienpont et al. found that the position of chromosome 6q24-q25 is related to human congenital heart disease. Further research found that two missense mutations in the TGF-β-kinase 1 / MAP3K7 binding protein 2 gene (TAB2) are activated heart failure

[0013] When the high-fidelity DNA polymerase is performing DNA amplification, when the primers are completely matched with the template, the polymerization reaction will be carried out directly in the polymerization center of the polymerase; when the primers are not completely matched with the template, it will be sent to the enzyme center for correction. The corrected primers are sent back to the polymerization center for the polymerization reaction; if the primers cannot be corrected immediately, the PCR reaction cannot proceed normally, thereby triggering the immature termination of the polymerization reaction

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