ABSTRACTS – ORAL PRESENTATIONS - AMCA, spol. s r.o.

We have tested optimal composition and performance of the screening tubes over 4
modifications of 8-color panel on the total of 270 peripheral blood samples from patients
and healthy donors. Of them, there were 29 patients with molecularly defined mutation
and 56 patients with Common variable immunodeficiency (CVID). We have reached
concordance with standard 4-color tubes. Major perturbations (absence of lymphocyte
subset) were correctly detected in Severe combined immunodeficiency (SCID) and
Cartilage-hair hypoplasia (CHH). Maturation perturbations, such as absence of memory
B cells or memory T cells were detected in Hyper IgM syndrome (HIGM), Wiskott-Aldrich
syndrome (WAS), DiGeorge syndrome (DGS) and CVID. Dedicated 8-color tubes focused
on maturational stages of T or B cells further investigated individuals with abnormal
results of the EuroFlow PID screening tube.
EuroFlow PID screening tube allowed us to detect abnormalities in all major lymphocyte
subsets. Standardization of the sample preparation protocols and data acquisition allows
for multi-centric evaluation of the rare diseases and will also enable sharing anonymized
phenotype data to facilitate diagnostic process and expand our understanding of cellular
immunity in PID.
Acknowledgements: Supported by grant NT/13271, NT/13287-4, NT/11414-5
P47. THE USE OF FLOW CYTOMETRY IN THE SELECTION OF PROBIOTIC BACTERIA
Dagmar Mudronova, Radomira Nemcova, Dana Ryznerova
Department of Microbiology and Immunology, University of Veterinary Medicine and
Pharmacy, Kosice, Slovak Republic; mudronova@uvlf.sk
Flow cytometry is mostly used for identification, counting and testing of some properties
of eukaryotic cells. In recent years, however, increasingly being used in microbiology,
especially for counting and viability testing of bacteria. Any potentially successful probiotic
bacteria designated for oral administration must fulfill some selection criteria. They
must be able to survive and grow in the gastrointestinal tract conditions and to adhere
to the mucosa of the gut. It is also necessary to respect the origin of the strain used and
its ability to inhibit pathogens. The strain should be precisely identified, safe, genetically
stable, it should have good growth properties, to maintain its high viability at processing
and when in storage. Depending on the desired outcome, a probiotic strain may need to
have additional properties, such as anticarcinogenic or hypocholestrerolemic effects, or
the ability to improve lactose utilization (Nemcová, 1997). In the first phase of selection,
most of the properties are tested in in vitro conditions. For tests on the gastrointestinal
tract and technological process survival are often used conventional, time- and materialconsuming
microbiological methods. An alternative, rapid and reliable technique for
viability assessment seems to be flow cytometry (Bunthof et al., 1999). The aim of this
study was to investigate the survival of beneficial bacteria in simulated gastrointestinal
conditions and during the technological process by flow cytometry.
For the simulation of the gastrointestinal tract conditions was prepared artificial gastric
juice with pH adjusted to different levels, intestinal juice and bile salts solutions with
Analytical Cytometry VII 143