Mutation analysis in the F8 gene in 485 families with haemophilia A and prenatal diagnosis in China

Background Haemophilia A (HA) is an X-linked bleeding disorder caused by mutations in the coagulation factor VII(F8) gene. Its incidence in men is estimated to be approximately 1/5000. Objective This study aimed to characterize the mutation spectrum of the F8 gene in 485 Chinese families, encompassing all HA phenotypic classes. Additionally, we evaluated the accuracy of prenatal diagnosis of foetuses at risk of having HA. Methods Long-Distance PCR (LD-PCR) and Multiplex PCR were used to detect inversions, next-generation sequencing (NGS) was used for point mutations, and multiplex ligation-dependent probe amplification (MLPA) was used for large deletions or duplications. Results A mutation spectrum of 478 HA families was produced. Throughout 26 exons and 15 introns, a total of 237 different alterations of mutations were detected, of which 146 are known mutations (64.5%) and 91 are novel mutations (35.5%). Prenatal diagnosis revealed 97 normal males (35.79%), 103 HA males (38.01%), 36 normal females (13.28%), and 38 HA carrier females (14.02%). Conclusion Using a systematic approach comprised of three steps, 237 pathogenic variants in 478 out of 485 patient samples (98.6%) were detected, including the identification of a heterogeneous mutation spectrum of 91 novel mutations. In addition, prenatal diagnosis of HA in pregnant carriers allowed for accurate determination of the foetal F8 gene state.