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The aim of this study was to evaluate and compare two genotyping methods for the examination of genetic diversity in C. jejuni and C. coli strains isolated from poultry. Moreover, the molecular identification of putative virulence factors was additionally included to establish a correlation between the isolates. Both typing methods, RAPD-PCR and ERIC-PCR, were found to have a high discriminatory power, D=0.95 and 0.94, respectively. The isolates were clustered to 28 groups by RAPD-PCR and to 29 by ERIC- PCR. Sometimes the ERIC or RAPD assays were able to further discriminate the isolates, which had the same virulence profile generated by the second technique. These PCR-based differentiation methods demonstrated the potential for initial rapid epidemiology studies, and when used in conjunction with distribution of virulence markers provide a high resolution strategy for accurately defining subtypes of Campylobacter.
Background. A tendency to venous thromboembolism is otherwise called hypercoagulability or thrombophilia. This disease can be acquired or have a genetic background, and may lead to pulmonary embolism. The basis for analysis and selection of treatment is genetic diagnosis, which detects the G1691A mutation in the factor V gene (factor V Leiden) – the best known congenital thrombophilia marker. Material and methods. The study was carried out in the years 2015-2017 on samples taken from patients (462 men and 1284 women) with a tendency to venous thromboembolism. Real-Time PCR was used to detect G1691A mutation in factor V gene. The analyses were performed in the Hematology Laboratory of the Center of Laboratory Medicine at the Medical University of Gdańsk. Results. Significant differences in the frequency of Leiden mutation were shown. This mutation predominated in men (25%), while in women G1691A mutation was detected with a 15% frequency (p=0.04). All possible genotypes were found among the subjects and the percentage of heterozygotes and homozygotes in both genders was similar. Conclusions. Congenital t hrombophilia a ssociated w ith G1691A mutation of f actor V Leiden gene was found to be more common in men than in women. All possible genotypes were determined in the pool of test subjects. The mutation was most frequently detected in patients between 30 and 40 years of age, and rarely after 70 years of age.
In search of an effective DNA typing techniaue for Acinetobacter baumannii strains for hospital epidemiology use, the performance and convenience of a new target seauence was evaluated. Using known genomic seauences of Acinetobacter baumannii strains AR 319754 and ATCC 17978, we developed single-locus variable-number direct-repeat analysis using polymerase chain reaction-restriction fragment length polymorphism (DR-PCR/RFLP) method. A total of 90 Acinetobacter baumannii strains isolated from patients of the Clinical Hospital in Bydgoszcz, Poland, were examined. Initially, all strains were typed using macrorestriction analysis of the chromosomal DNA by pulsed-field gel electrophoresis (REA-PFGE). Digestion of the chromosomal DNA with the Apal endonuclease and separation of the fragments by PFGE revealed 21 uniaue types. Application of DR-PCR/RFLP resulted in recognition of 12 clusters. The results showed that the DR-PCR/RFLP method is less discriminatory than REA-PFGE, however, the novel genotyping method can be used as an alternative techniaue for generating DNA profiles in epidemiological studies of intra-species genetic relatedness of Acinetobacter baumannii strains.
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