European Human Genetics Conference 2007 June 16 – 19, 2007 ...

Genomics, technology, bioinformatics
Application has a very strong image processing mechanism and flexible
database architecture. Medical staff puts tests into scanner and
GENIUS gets image from device. Then application checks the test
type. After the identification of test, it searches mutation areas and
other critical check points on the stripassay. By using analysis results,
GENIUS refers to database and generates comments about the results.
Adding a new test to system takes approximately 2-4 hours depending
on the number of mutations on the stripassay. So it is very
easy to adapt GENIUS for different markets which use different stripassays.
Oracle 10g is the database of the GENIUS. Addition to this Oracle BI
EE is being used for analytic reporting.
P1298. Comparison of two fluorescent dsDNA binding dyes
SYBR Green I and EvaGreen for Melting Curve Analysis
J. Radvansky1 , G. Minarik1 , A. Ficek1,2 , P. Resko1 , L. Kadasi1,2 ;
1 2 Comenius University, Faculty of Natural Sciences, Bratislava, Slovakia, SAS,
Institute of Molecular Physiology and Genetics, Bratislava, Slovakia.
One of the basic detection systems for real-time PCR is based on ds-
DNA binding fluorescent dyes, from which SYBR Green I is the best
known. However, the SYBR Green I is not the only dye for these applications.
Because of their unique feature (enhanced fluorescence when
bound to the dsDNA) SLDs (SYBR Green Like Dyes) are frequently
used in detection of specific PCR products by melting curve analysis
(MCA). Recently, a new fluorescent dye EvaGreen has appeared
which shows (by the manufacturer’s information) better properties for
real-time PCR than SYBR Green I. To investigate the more appropriate
dye for our MCA applications we decided to compare some properties
of these two dyes such as photostability, detection limit with diverse
range of dye concentration and PCR product concentration and some
parameters of the final peak in MCA. Since the molar concentration
of each dye is different in the 1x working solution our measurements
were done by the basic working concentrations and by equal molar
concentrations too. We also compared the accuracy of the Tm values
obtained from a larger set of each sample. In our study we used 4
PCR products (for the detection of the 35delG, W24X, R127H mutations
and a polymorphism GJB2-SNP1 in GJB2 gene). According
to our findings EvaGreen has better properties (the peaks obtained
from MCA by EvaGreen are generally higher and sharper and the Tm
values are more accurate) however it shows lower photostability than
SYBR Green I. Detailed results will be presented on our poster.
P1299. Association study of therapeutic response and SERT,
MDR1 and 5-HT2c gene polymorphisms in female schizophrenic
patients
N. Bozina 1 , J. Sertic 1 , V. Medved 2 , M. Rojnic Kuzman 2 , L. Hotujac 2 ;
1 Center for Functional Genomics and Clinical Institute of Laboratory Diagnosis,
Zagreb University Hospital Center, Zagreb, Croatia, 2 Department of Psychiatry,
Zagreb University Hospital Center, Zagreb, Croatia.
Interindividual differences in treatment response to second generation
antipsychotics point out that genetic factor may be relevant. The aim
of this study was to investigate the relationships between variants of
serotonin transporter gene (SERTPR and SERTin2), serotonine receptor
(5-HT2c-759C/T) and multidrug resistant gene (MDR1-2667G/
T, 3435C/T), and initial symptomatology and treatment response in
106 female schizophrenic patients treated with olanzapine for up to
3 months. Afterwards, we compared allele, genotype and haplotype
distributions between those patients and 108 control female subjects.
Methods: Genotyping was performed by PCR-RFLP, and real-time
PCR methods. To assess and evaluate therapeutic response, all patients
were rated using PANSS. Overall, the presence of SERTPR-S
allelic variant and SS genotype was associated with significantly more
weight gain in subjects who were non-obese at the time of admission
(p= 0.02). The presence of SERTPR-L variant was associated with significantly
better treatment response measured with total PANSS and
general PANSS subscale (p A), MTHFR
(677C> T), factor V Leiden (1691FVG> A), PAI-1 (5G> 4G), GPIIIA
(196C> T), fibrinogene (455G> A) genes which result in hereditary
thrombophilia. We have developed the fast and easily adapted method
of detection mutations associated with thrombophilia, suitable for
technological process at any standart molecular diagnostic laboratory.
This convenient and reliable detection of mutations is based on multiplex
PCR with subsequent hybridization on the biochip. Use of this
technique open new opportunities for carrying out large-scale population
researches of genetic predisposition to thrombophilia. The practical
recommendations have been developed for the choice of therapy
strategy in case of investigated mutations detection.
P1302. A homogeneous high-throughput assay for the primary
screening of type 1 diabetes related HLA-DQB1 alleles
M. Kiviniemi 1 , J. Nurmi 2 , J. Ilonen 1,3 , T. Lövgren 4 ;
1 Immunogenetics Laboratory, University of Turku, Turku, Finland, 2 Abacus Diagnostica
Oy, Turku, Finland, 3 Department of Clinical Microbiology, University
of Kuopio, Kuopio, Finland, 4 Department of Biotechnology, University of Turku,
Turku, Finland.
We have developed a homogeneous genotyping system allowing
simple, low-cost and rapid genotyping of thousands of samples. The
method utilizing an asymmetric PCR and subsequent hybridization of
allele specific probes with LNA additions for the genotyping of HLA-
DQB1 alleles *02, *0302 and *05/6 can be used as the primary screen-