0-TESTO COMPLETO.pdf - Fondazione Santa Lucia

TEL.13 – Clinical and laboratory criteria for FSHD diagnosis in view of a national registry…
patients will be examined and will receive a FSHD score. Our intent is to identify
the correlations, if any, between the genetic rearrangement and factors,
which may influence the severity of the disease. The uniformity of the different
centers will be guaranteed by a preliminary meeting organized by the clinical
coordinator. All clinical data will be introduced in the specifically
designed database.
B. Molecular analysis – Chromosomal DNA will be prepared from isolated
lymphocytes according to the standard procedure.
Restriction endonuclease digestion of DNA will be performed in agarose
blocks with the appropriate restriction enzyme: EcoRI and BlnI (p13E-
11probe), Hind III (4qA/4qB probes), XapI (chromosomal assignement of
p13E-11 alleles), Tru9I (D4Z4 repeat probe), NotI (B31 probe). DNA will be
separated by Field Inversion Gel Electrophoresis (FIGE) and Pulsed Field Gel
Electrophoresis (PFGE). Probe hybridization will allow to analyze the appropriate
genomic region.
Data processing. A total of 1752 patients are currently available. The patients
and their families will be summoned through telephone calls or mail. We
expect a response rate of 65-75% throughout the entire funding period. This
will ensure an enrolment of 1138-1314 patients. We expect that an equal number
of relatives will be also enrolled. The patient code will allow us to process
both the clinical and personal data, fulfilling the current laws on privacy. For
data protection, forms will be numbered before being sent to the Coordinator.
Each partecipating center will be identified by a prefix number, from 01 to 13.
Each form will be numbered in ascending order like: 01/001, 01/002, 01/003.
etc. Each numbered card will be assigned to a given patient.
A preliminary informative meeting will be held to discuss and illustrate
the forms to the physicians of all the collaborating centers. This will guarantee
the use of a standardized methodology to be applied during the patients’
examination. Subsequent meetings will be organized to discuss all the peculiar
cases. Information collected in the appropriate forms will be inserted into
the specific database. Statistical analyses to estimate the prevalence of the disease
in the Italian population, the relative frequency of sporadic and familial
cases, and the estimation of the risk of relatives of affected recruited patients
will be performed under the supervision of an expert statistician.
FSHD score will be used to describe in a single parameter the patients’
phenotype. Comparison of risk of severe symptoms (higher FSHD score)
between patients with different fragment size will be estimated by using logistic
regression adjusted for age. Correlations will be tested by Spearman’s rank
correlation test.
To assess the hypothesis of significant difference between two groups (e.g
compound heterozygotes and single deletion carriers) for given variables
(muscles involved, associated polymorphisms 4qA/4qB, etc) the non-parametric
Mann-Whitney t-test will be used. All analyses will be performed with
STATA software Version 8 (Stata Corporation, College Station, Texas).
Statistical analysis will be used to validate a standard diagnostic protocol
for FSHD.
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