Amplification primer, kit and method for detecting f8 gene mutation

Abstract

The invention discloses amplification primers, a kit and a method for detecting F8 gene mutation, and belongs to a gene detection technique. Sixty-five pairs of the PCR primers are designed through sequence information of an F8 gene, PCR targeted amplification is performed on a target area of the F8 gene, and the amplified area covers 99.5% of an exon encoding area of the F8 gene. According to the method, multiple PCR targeted amplification is combined with an Ion torrent PGM high-throughput sequencing technique, simultaneous and parallel detection of multiple samples is achieved, the detection range of F8 gene mutation is widened, the detection throughput of F8 gene mutation is increased, the detection cycle is shortened, and the cost is reduced.

Description

technical field [0001] The invention belongs to the field of in vitro gene detection, in particular to an amplification primer, kit and method for detecting F8 gene mutation Background technique [0002] Hemophilia (hemophilia) is a bleeding disorder of inherited coagulation dysfunction. Due to gene mutations in coagulation factors, the concentration of coagulation factors decreases or is lacking, resulting in prolonged coagulation time and a tendency to bleed after minor trauma. According to the lack of coagulation factors, clinically divided into hemophilia A (hemophilia A, HA) and hemophilia B (hemophilia B, HB). Its clinical manifestations are mainly spontaneous bleeding after minor trauma or severe bleeding after trauma and surgery. Bleeding sites are usually found in large weight-bearing joints (such as knees, elbows, ankles, wrists, iliac, shoulders, etc.) and muscles / soft tissues, and may also manifest as visceral bleeding (such as intraperitoneal, retroperitoneal, ...

AI Technical Summary

Problems solved by technology

At present, direct Sanger sequencing of PCR products is a commonly used F8 gene detection method. Although this method is more accurate, the sequencing workload is huge, the efficiency is low, and the requirements for operators are high, which limits the large-scale clinical application of this method. Can only be carried out in qualified laboratories

[0003] Currently, CN102230002A discloses a kit for detecting whether the hemophilia pathogenic gene F8 gene and F9 gene are mutated, but it needs to use 24 p

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