title: Prenatal diagnosis in haemophilia A: experience of the genetic diagnostic laboratory.
abstract: The paper describes the experience of the Genetic Diagnostic Laboratory in prenatal testing for haemophilia A, an X-linked recessive disease caused by mutations in the F8 gene. Knowledge of a familial mutation prior to pregnancy can benefit prenatal diagnosis and decrease wait time for molecular testing during pregnancy. This is a retrospective review of a series of pregnant women who pursued F8 gene testing from December 1997 through May 2012, highlighting three cases, which demonstrate the technical complexities of analysis and the implications of not knowing carrier status prior to pregnancy. Mutations of the F8 gene were detected in affected males, obligate female carriers and suspected female carriers by DNA sequencing, inverse-PCR, qRT-PCR, Southern blot and exonic dosage analysis. The same methods were used to analyse prenatal samples from obligate or suspected female carriers upon request. Maternal cell contamination studies were performed for all prenatal samples analysed. Ninety-nine women pursued F8 testing during pregnancy, either for carrier status alone or carrier status and prenatal diagnosis. Ninety-one women (91%) requested carrier testing because they did not know their F8 mutation carrier status prior to pregnancy. Eight women requested prenatal diagnosis only, and only 4 of these were aware of their mutation status. Thirty-seven individuals were found to be mutation carriers. Forty-two prenatal samples were received for prenatal diagnosis. In total 21 foetuses were identified as mutation carriers. Mutation detection was complex and increased the turnaround time in some cases. Only four of 99 women who submitted samples for F8 testing were aware of their F8 mutation status prior to pregnancy. Knowledge of F8 mutation status prior to pregnancy allows for efficient prenatal diagnosis, when desired. Thus, preconception genetic counselling is required to inform patients of the available options and the complex and time-consuming nature of F8 testing.
authors.affiliation: The Genetic Diagnostic Laboratory, Department of Genetics, Perelman School of Medicine, The University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: L
authors.initials: L
authors.lastname: Kessler
authors.forename: R
authors.initials: R
authors.lastname: Adams
authors.forename: L
authors.initials: L
authors.lastname: Mighion
authors.forename: S
authors.initials: S
authors.lastname: Walther
authors.forename: A
authors.initials: A
authors.lastname: Ganguly
mesh_terms: D000328:Adult; D030541:Databases, Genetic; D005169:Factor VIII / Q000235:genetics; D005260:Female; D005820:Genetic Testing; D006467:Hemophilia A / Q000175:diagnosis* / Q000235:genetics*; D006801:Humans; D008297:Male; D009154:Mutation; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods; D012189:Retrospective Studies
chemical_list: D005169:Factor VIII
keywords: factor VIII mutations; genetic counselling; haemophilia A; prenatal diagnosis