Portada Simposios - Supplements - Haematologica

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RECENT ADVANCES
IN THE MOLECULAR DIAGNOSIS
OF HEMOPHILIA
F. VIDAL AND D. GALLARDO
Unitat de Recerca. Centre de Transfusió i Banc de Teixits
de Barcelona.
Characterization of hemophilic patients at the
molecular level, particularly of those affected by hemophilia
A, has been hampered by the complex
structure and the large size of the genes involved as
well as by the high frequency of new mutations. In
addition, existing nucleotide sequencing techniques
are relatively difficult to establish on a routine basis
and remain as an expensive and time-consuming option.
A direct consequence of this situation is that,
on the brink of the 21 st century, many hemophilic
patients cannot benefit from a definitive and precise
molecular diagnosis to explain the ultimate reason
for their pathology.
Accurate molecular characterization is essential
for this collective in order to perform decisive genetic
counseling for female carrier detection and a reliable
prenatal diagnosis. Additional benefits derived from
the knowledge of a specific mutation include information
on the side effects of replacement therapies
with recombinant factors, such as the risk of inhibitor
formation, to select patients for different treatments
or follow-ups. Furthermore, identification of
the causative mutation is necessary when seeking approval
of human clinical trials in oncoming strategies
of gene therapy.
To circumvent the direct sequencing approach several
laboratories have developed a number of mutation
screening techniques, all of which have a final
common step: nucleotide reading. Unfortunately,
these techniques are very labor intensive and
require a highly qualified staff to be performed on a
routine basis. For the time being, until easier and
more feasible alternatives are available, linkage
analysis with indirect markers is still the standard
practice for genetic counseling and prenatal diagnosis
in hemophilia. This method is straightforward
but has some important limitations: homozygosity,
higher risk of recombination when only extragenic
markers are informative, unavailability of key individuals
within a family and, most importantly, sporadic
cases without any previous history of hemophilia
(up to 50 % of families).
Recent advances in molecular biology, engineering
and bioinformatics have make possible the development
of new protocols and related equipment for
exhaustive molecular diagnosis which, in terms of
sensitivity, rapidity and simplicity, were unthinkable
just a few years ago. Because linkage analysis does
not identify the mutation that causes the defect, an
optimized direct sequencing approach should be
contemplated for genetic counseling and prenatal