Kit for detecting one pathogenic mutation site in male infertility Kif18a gene, and PCR amplification method thereof

Abstract

The present invention discloses a kit for detecting one pathogenic mutation site in male infertility Kif18a gene, and a PCR amplification method thereof. The method is characterized in that mutation detection primers resisting 3'-5'exonuclease digestion modification are introduced into a high fidelity DNA involved PCR reaction so as to improve the PCR specificity, and the product band of the gel electrophoresis can be directly observed so as to determine the genotype of the corresponding mutation site of the Kif18a gene. According to the present invention, the detection kit has characteristics of low instrument requirements, simple operation, economy, no requirement of sequencing, and capability of being operated in general molecular biology laboratories.

Description

technical field [0001] The invention belongs to the clinical detection technology in the field of biomedicine, and relates to a detection method for a pathogenic mutation site of a human male infertility gene. [0002] The invention provides a PCR kit for detecting mutations of human male infertility pathogenic genes. The content of the present invention relates to a kit for detecting a pathogenic mutation site in the Kif18a gene of male infertility and its PCR amplification method, and the detection result can be used for clinical auxiliary detection of male infertility. Background technique [0003] Male infertility refers to the fact that a man fails to impregnate his healthy spouse within one year when he has regular sex life and does not use contraception. Male infertility (Male Infertility) seriously affects modern men's health and family happiness. According to WTO statistics, about 15% of couples of childbearing age are infertile, of which male factors account for 5...

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Problems solved by technology

[0016] 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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