[Study on genetic diversity of genes encoding main adhesins among nontypeable Haemophilus influenzae strains isolated from children in Poland]. / Badania profilu determinant genetycznych kodujacych glówne adhezyny bezotoczkowych szczepów Haemophilus influenzae izolowanych od dzieci w Polsce.

The study was based on hypothesis that in the nontypeable population of H. influenzae strains isolated from children there are some genetically predisposed to induce symptomatic infection in children and that they might be divided into different groups depending on profiles of genes encoding main adhesins synthesis. The work aimed at analysis of distribution of genes encoding adhesins and evaluation of domination possibility of some strains representing particular adhesins genes profiles among NTHi population. Results of the study revealed that among population of NTHi strains, distribution of genes encoding main adhesins are differing. Among children, NTHi strains harbouring genes encoding HA and HMW1/HMW2 adhesins were more prevalent in healthy children and in children with symptomatic infections, respectively. Analysis of strains harbouring main adhesins profiles might be a useful screening method in monitoring strains circulating among children, in order to determine the most invasive NTHi strains.

[Diagnostic difficulties in identification of Haemophilus influenzae colonising the nasopharynx of children]. / Problemy diagnostyczne przy identyfikacji szczepów Haemophilus influenzae kolonizujacych jame nosowo-gardlowa dzieci.

The frequency of capsulated or non-capsulated Haemophilus influenzae strains colonisation among children attending day-care centres or orphanages has been studied. Detection of capsulated or non-capsulated H. influenzae strains has been compared for agglutination test and PCR. Misdiagnosing of H. influenzae serotype with agglutination found in the study suggest that the frequency of Hib strains colonizing the nasopharynx might be lower that previously evaluated. Due to perspectives of the wider use of Hib Immunisation in the future, more efficient diagnosis scheme for identification/differentiation of capsulated and non-capsulated H. influenzae strain should be undertaken.

[Transmission of Haemophilus influenzae type b strains among children attending day-care centers and orphanages]. / Badania transmisji szczepów Haemophilus influenzae typu b krazacych w populacji dzieci w przedszkolach i domach dziecka.

Goal of the work was to evaluate the differences level among H. influenzae strains of b serotype isolated from children attending day-care centres and orphanages and among strains isolated from invasive infections. In the work PFGE in Hib strains transmission examination and for epidemiological studies among three sources of invasive infection was applied. Among 35 Hib strains tested and control strain 8 different pulsotyped were found. Among 21 strains colonising the nasopharynx of healthy children, and among 13 Hib strains isolated from cerebrospinal fluid, 6 and 1 pulsotypes were found, respectively. Results obtained show that healthy children might be colonizing with genotypes characteristic for Hib strains isolated from invasive infections. In this view wider Hib vaccination seems be expected, as Hib circulation is common.

Amplified fragment length polymorphism (AFLP) versus randomly amplified polymorphic DNA (RAPD) as new tools for inter- and intra-species differentiation within Bordetella.

Automated amplified fragment length polymorphism (AFLP) and randomly amplified polymorphic DNA (RAPD) techniques with fluorescently labelled primers were used to track differences among isolates of the eight known species of the Bordetella genus. Eighty-one representative strains of these species from international and Polish bacterial collections were genotyped according to RAPD protocols using primer 1254 or 1247, and AFLP involving EcoRI/MseI or newly designed SpeI/ApaI restriction/ligation/amplification procedures. By comparing AFLP and RAPD data, it was concluded that the discriminatory power of AFLP is higher in comparison with RAPD for both intra- and inter-species differentiation of isolates of the Bordetella genus. The most precise level of inter-species discrimination and the highest level of intra-species discrimination of the Bordetella isolates of the eight species were observed in the AFLP EcoRI/MseI and SpeI/ApaI sets, respectively. Both techniques might provide alternative tools for the identification of Bordetella at the genomic species and strain levels, and thus may be valuable in human and veterinary diagnostics as well as in epidemiology. By applying the AFLP technique presented in this article, more precise data on the emergence of newly acquired and/or on expanded clones and transmission routes of isolates of the Bordetella genus in the human and animal environments might be obtained.

[Polymorphism analysis of Haemophilus influenzae strains isolated from healthy children with respect to amplification reaction]. / Badanie polimorfizmu genetycznego szczepów Haemophilus influenzae izolowanych od zdrowych dzieci z wykorzystaniem reakcji amplifikacji.

The random-amplified polymorphic DNA (RAPD) assay was used to generate DNA fingerprints for 44 isolates H. influenzae obtained from healthy children. Problems with reproducibility and discriminatory power, frequently cited in the literature, were overcome by optimization procedure allowing to achieve reliable conditions for H. influenzae analysed. Particular parameters of RAPD fingerprinting were evaluated with respect to selection of best working primer, DNA polymerase and DNA concentration for amplification pattern. This study proved high sensitivity and efficiency of optimized RAPD profiling applicable for searching the epidemiology traces.