2008 Barcelona - European Society of Human Genetics

Genetic counselling, education, genetic services, and public policy
made available . New ways to involve the public in policy decisions on
screening and testing are necessary .
P09.67
the importance of genetic counselling for parents with normal
constitutional karyotypes having multiple spontaneous
abortions with aneuploidy
V. Culic1 , J. Pavelic2 ;
1 2 Pediatrics Clinic, Department for Medical Genetics, Split, Croatia, Institute
Rudjer Boskovic, Molecular Genetics, Zagreb, Croatia.
The purpose of this research is to establish the hypothesis that two hits
are responsible for aneuploidy which results in spontaneous abortion .
The first hit could have been triggered by patient’s former diseases
of urinary and/or genital system . The number of spontaneous abortions
recorded in their siblings of second generations creates predisposition
that may end with termination of pregnancy . The second hit
is usually triggered by chronic presence of EBV virus in male or/and
female partner. It has been demonstrated that there is no significant
relation between the age of male or female partners and the number
of spontaneous abortions . Both males and females had former urinary
or/and genital infections and the number of spontaneus abortions
among the siblings of second generation is twice as high in relation
to average population . Variations in constitutional karyotypes (per inv
9, 9qh+) are also evident as well as presence of antibodies for EBV,
CMV, or HSV2 viruses . Reactivation of latent EBV infection provides
possibility for second hit . Aneuploidy or tetraploidy is found at 23% of
samlpes . Therefore we think that the presumed hypothesis of two hits
is clearly established . Partners with spontaneous abortion with and
without chromosomal abnormalities must undergone genetic counselling
process for preventing trigger/s or to repeat the proces, because
spontaneous abrotion is an multifactorial disease .
P09.68
milestone in teaching genetics
R. Dragotoiu;
Medical and Pharmacy University “Carol Davila”, Bucuresti, Romania.
On the verge of a new legislation in the educational system, when one
of the main goals is the alignment to the international standards, it is
suggested that student’s evaluation has to become mandatory and its
results should be made public by the university . More than a decade
ago I evaluated my teaching methods and presented the results at the
ESHG meetings . Three years ago I decided to change the way I taught
the practicals .
Questionnaires were answered by 76 first year students of the Medical
and Pharmacy University “Carol Davila” to evaluate the quality of the
practicals they attended during the first term in the Department of Genetics
. They were questioned about the quantity of information versus
amount of time, the quality of information and presentation and if they
enjoyed the practicals .
Answers to the question: Have you been satisfied with the way the
practicals unfolded?
very much 7%
a lot 42%
enough 50%
satisfactory 1%
almost not at all none
not at all none
For several years I have stopped asking students for an evaluation, because the results
were the same: almost half in favor of frontal teaching and the other half preferring
the interactive method. Since I have modified my teaching method, the speed and
the amount of information also changed, but only 7% of students considered it was
too much data.
Adapting the teaching method to the students’ present needs makes them more prone
to enjoy the practicals. The study did not focus on achieving genetic knowledge, but
apparently this did not improve.
P09.69
thrombophilic gene mutations: two different approaches to the
validation of commercial and “in-house” DNA diagnostic assays
A. Stambergova, J. Camajova, M. Macek Jr.;
Department of Biology and Medical Genetics, Prague 5, Czech Republic.
Validation of molecular genetic analytical methods is the basic tool for
quality assurance and has to be part of routine practice in clinical-diag-
nostic laboratories that are working in compliance with the ISO15189
guidelines (www .iso .org) . Moreover, OECD has issued “Guidelines for
Quality Assurance in Molecular Genetic Testing (www .oecd .org\sti\biotechnology)”
which stipulate that laboratories providing clinical services
should be working within a “Quality Assurance System” . Via method
validation laboratories assess assay performance and demonstrate
their suitability for the intended purpose . It is up to individual laboratories
to establish an acceptable validation protocol in order to comply
with general ISO principles . Here we report practical examples of validation
procedures utilized for an ,,in-house“ test that we developed for
the detection of thrombophilic mutation PAI-1, versus approaches applied
to a commercially produced RHA Kit Thrombo (Labo Bio:Medical
Products; CE/IVD marked) . For examination of the thrombophilic mutation
PAI-1 our laboratory uses an “in-house” method based on fragment
analysis of restriction endonuclease- digested amplicons, while
the commercial assay is based on reverse slot hybridization (RHA)
for simultaneous detection and identification of mutations in the genes
coding for FII G20210A, FV-Leiden, including the MTHFR 677C/T variant
. In both instances we assessed basic validation parameters including
specificity, sensitivity, repeatability, reproducibility and robustness.
Differences between both validation protocols are within the scope of
validation parameters being tested . These procedures enabled us to
confirm diagnostic reliability of both techniques and provide practical
examples for other laboratories validating their genetic tests . Supported
by VZFNM00064203(6112) and EuroGentest .
P09.70
Genotyping of mtHFR 677c>t and 1298A>c variants by high
resolution melting of small amplicons: an example of method
validation.
P. Norambuena, J. Copeland, P. Krenkova, M. Macek Jr., A. Stambergova;
Department of Biology and Medical Genetics, Prague 5, Czech Republic.
High Resolution Melting (HRM) is a simple, cost-effective and rapid
mutation scanning method . By reducing the respective amplicon size,
it is possible to increase the resolution for reliable discrimination between
homozygous wild-type and homozygous mutant samples . We
introduced HRM close-tube method in our laboratory for genotyping of
the most common variants in the MTHFR gene: 677C>T (rs1801133:
C>T) and 1298A>C (rs1801131: A>C). PCR amplification and HRM
was performed on the LightCycler® 480 Real-Time PCR System
(Roche). For validation purposes we evaluated specificity, sensitivity,
repeatability, reproducibility and the ability of the method to perform
reliable calls . We were able to perform unambiguous calls in 98 .4%
(n=381) of the cases for rs1801133: C>T and in 98 .1% (n=104) of the
cases for rs1801131: A>C . For the 6 (1 .6%) and 2 (1 .9%) unknown
samples for rs1801133: C>T and rs1801131: A>C respectively, by repeating
the analysis we reached 100% of calls . Since in the end all
calls were reliably detected by the LightCycler software we reached
100% specificity and sensitivity. Repeatability and reproducibility were
also consistent . In conclusion, after optimization and diagnostic validation
HRM of small amplicons is a reliable and precise method that can
be broadly applied in DNA diagnostics of other genes .
Supported by VZFNM00064203(6112) and EuroGentest .
P09.71
training on quality management to support genetic testing
laboratories in continuous improvement and progress towards
accreditation
M. A. Morris 1,2 , S. Berwouts 3,2 , A. Wynants 4,2 , E. Dequeker 3,2 ;
1 Laboratoire de Diagnostic moléculaire, Service de Médecine Génétique, University
Hospital, Geneva, Switzerland, 2 EuroGentest Network of Excellence,
Leuven, Belgium, 3 Centre of Human Genetics, University of Leuven, Leuven,
Belgium, 4 Management Consulting & Research, Heverlee, Belgium.
ISO 15189:2007, the international standard developed specifically for
medical laboratories, requires that staff receives relevant training in
quality assurance and quality management (5 .6 .1) . In response to the
growing interest in accreditation and quality improvement, a series of
interactive two-day training workshops were organized for genetics
laboratories, within the framework of the European project EuroGentest
.
The subjects of these workshops varied from living with quality systems
and comparing the different standards for accreditation, to specific
topics such as internal audit, managing the human side of change