A novel deletion mutation in the F8 gene in hemophilia A
Objective This study aims to identify the pathogenic gene mutation in a family with hemophilia A by employing a comprehensive approach using long-distance PCR,multiplex PCR techniques,high-throughput sequencing,and multiplex ligation-dependent probe amplification(MLPA),and to provide prenatal counseling for the family.Methods The study used long-distance PCR(LD-PCR)and multiplex PCR to detect inversion mutations in introns 22 and 1 of the F8 gene;whole exome sequencing(WES)to detect point mutations,minor insertions,or deletions in the F8 gene,with validation by Sanger sequencing;and MLPA to detect large fragment deletions/duplications in the F8 gene.Subsequently,first-generation sequencing was employed to verify the genetic mutations in other blood relatives within the family,and software analysis was conducted to evaluate the potential impact of this mutation on the protein structure.Results In the proband,a new mutation c.5682del(p.Glu1894Aspfs51)in the F8 gene was identified,with no other point mutations,insertions,deletions,or inversions in introns 1 and 22 observed.This mutation was validated in other family members,showing consistency between the disease phenotype and the genetic mutation segregation.Conclusion The novel mutation c.5682del(p.Glu1894Aspfs51)carried by the mother is the cause of hemophilia A in the child of this family.
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