Experience of a single Italian center in genetic counseling for hemophilia: from linkage analysis to molecular diagnosis.
OBJECTIVE
We describe our three year experience in genetic counseling at the Castelfranco Veneto Hemophilia Center, Italy.


METHODS
A total of 258 individuals were involved in the study of 142 females. These formed 40 families with hemophilia A and 6 families with hemophilia B. Following pedigree analysis, the FVIII inversion was first examined in severe hemophilia A patients by polymerase chain reaction (PCR) analysis. DNA polymorphisms were used to track the affected gene through the remaining families. In uninformative cases, we initiated analysis of the FVIII or FIX gene coding region by conformation sensitive gel electrophoresis and DNA sequencing to identify the mutation responsible for the disease.


RESULTS
The FVIII gene inversion was present in 16 of the 32 patients (50%) affected by severe hemophilia A and was informative for 44 females. For hemophilia A, 45 cases (55%) were informative by linkage analysis, however 37 (45%) were uninformative because of lack of key individuals, homozygosity, or sporadic disease. Information from extragenic linked polymorphisms alone was present in 9 cases (6%). For hemophilia B, linkage analysis was informative in only 50% of females (8 out of 16). To date, nine mutations have been identified in patients with hemophilia A and three in patients with hemophilia B. Six novel missense mutations in hemophilia A are discussed briefly.


CONCLUSIONS
Using this approach we are now able to offer accurate genetic analysis to virtually all families with hemophilia.
Tagariello
G
G
Castelfranco Veneto Hospital, 31033 Castelfranco Veneto (TV), Italy. tagariello@ulssasolo.ven.it
Belvini
D
D
Salviato
R
R
Are
A
A
De Biasi
E
E
Goodeve
A
A
Davoli
P
P
D007446:Chromosome Inversion; D005164:Factor IX / Q000235:genetics; D005169:Factor VIII / Q000235:genetics; D005260:Female; D005817:Genetic Counseling / Q000379:methods / Q000592:standards; D008040:Genetic Linkage; D005820:Genetic Testing; D006467:Hemophilia A / Q000235:genetics*; D002836:Hemophilia B / Q000235:genetics; D006801:Humans; D007558:Italy; D008297:Male; D009154:Mutation; D010375:Pedigree
D005169:Factor VIII; D005164:Factor IX