[Sources of information and knowledge on childhood immunisations among Polish parents]. / Zródla informacji oraz wiedza rodziców na temat szczepien ochronnych w Polsce.

The aim of the study was to obtain information on parents' knowledge about mandatory and recommended vaccinations in Poland. Computer-assisted telephone interviews (CATI) method was used. The interviews were collected from parents who had children aged < or = 3 years. Two-stage sampling was used: first, a list of 3,000 households with children < or = 3 years was quota-selected from a consumer database collecting contact information from 95% mothers during deliveries; in the second stage, random digit dialling was used to attempt the interview with parents. The questionnaire applied was based on the survey developed by UK Department of Health. A total of 1045 interviews were collected, 960 from mothers, and 85 from fathers. The response rate was 77%. Mean age of parents was 28.9. Most parents (72%) had secondary or lower level of education. General practitioners (about 90% respondents) were found the most important, and the most reliable source of information for parents. Media (66%), including Internet (18%), were the second main source of theirs knowledge, but they were not considered as reliable source. Parental knowledge on obligatory vaccinations included in the childhood immunization schedule was found to be very low. Only 30% and 5% of respondents spontaneously recognized polio and Hib vaccines, respectively mandatory. The largest proportion of respondents indicated hepatitis B (46%) and tuberculosis (45%) as serious illness, and only 22% and 23% considered diphteria, and measles as serious diseases. Poor knowledge on childhood immunisations among Polish parents needs to be addressed by improvement of reliable information on vaccines and vaccine safety.

[Parental attitudes towards childhood immunisations in Poland]. / Postawy rodziców wobec szczepien ochronnych w Polsce.

The aim of the study was to obtain information on parents' attitudes towards vaccinations included in the childhood immunisation schedule. Computer-assisted telephone interviews (CATI) method was used. The interviews were collected from parents who had children aged three years old. Two-stage sampling was used: firstly, a list of 3,000 households with children < 3 years old was quota-selected from a consumer database collecting contact information from 95% mothers during deliveries. Random digit dialling was used to attempt the interview with parents. The 40-item questionnaire was based on the questionnaire developed by UK Department of Health. Overall, the perception of routine, mandatory immunization of children was positive. Only 17 parents (1.6%) refused the vaccination which had been offered, and 398 parents (38.0%) paid for a vaccine recommended for their child. In general, parents believed that immunisations were important for protecting the society against infectious diseases, although they found some problems in the way vaccines were delivered. Approximately half of respondents thought that vaccination against many diseases was harmful. In terms of perception of the risk related to vaccines parents were less confident in the currently introduced vaccines and those which protect against diseases rarely seen in the population. Pneumococcal vaccine was considered as risky by 27 persons (2.6%), and polio vaccine by 17 (1.6%). Greater concern about the safety of vaccines was expressed by older parents, residents of towns and highly educated individuals. Systematic monitoring of parents' attitudes towards vaccination would help to address public health actions more adequately.

[Occurrence of potentially invasive nontypeable Haemophilus influenzae strains isolating from children attending day-care centres and orphanages]. / Wystepowanie u dzieci z przedszkoli i domów malego dziecka potencjalnie inwazyjnych, bezotoczkowych szczepów Haemophilus influenzae.

Close contacts between children attending day-care centres, orphanages or similar institutions favours mutual transmission of infections with nontypeable H. influenzae (NTHi) strains. NTHi are transmitted via air-droplets or via direct contact with respiratory system exudates from nonsymptomatic carriers. The study aimed at monitoring of potentially invasive nontypeable H. influenzae strains of hmwA+ profile among children in day-care centres and orphanages. Monitoring of prevalence of strains of hmwA profile in a single day-care centres within 8 months confirmed high level of NTHi strains transmission including NTHi strains potentially invasive. It has been shown, that potentially invasive NTHi strains appear with different frequency in day-care centres and orphanages. It also points out that dissemination NTHi is easy in such an environment.

[Study of genetic diversity of nontypeable Haemophilus influenzae strains isolated from healthy children and patients with infection symptoms]. / Genetyczna zmiennosc bezotoczkowych szczepów Haemophilus influenzae izolowanych od nosicieli i chorych dzieci z objawami zakazen.

In the study the usefulness of genotyping methods for genetic variability examinations of non-typeable H. influenzae strains circulating in population as well as level the variability of NTHi strains isolated from healthy children and from symptomatic infection cases have been evaluated. Among genotyping methods evaluated, AFLP method of the MfeI/BglII set has been found most useful to study level of genetic variability of NTHi strains population. It has been shown that NTHi strains colonizing nasopharyngeal of healthy children present higher polymorphism level than strains isolated from patient with clinical symptoms of NTHi infection.

[Evaluation of whole-cell and acellular pertussis vaccines effectiveness in clearance of experimental B. pertussis infection in mice]. / Ocena efektywnosci pelnokomórkowych i bezkomórkowych szczepionek przeciw krztuscowi w eliminacji eksperymentalnego zakazenia myszy Bordetella pertussis.

The study is based on assumption that B. pertussis strains harbouring different allele variants of genes encoding subunit S1 of pertussis toxin and pertactin might be eliminated with different efficiency from lung tissue of mice which were immunized with whole-cell and acellular pertussis vaccines. It has been assumed that strains containing combinations of genes alleles which were not prevalent since 1990-ties are consisting of mutated strains in respect to pertussis toxin subunit S1 and pertactin, and are capable to decrease efficiency of pertussis vaccines. Experiments performed in vivo dealt with activity of tested vaccines against B. pertussis strains of different combinations of ptxS1/prn. The study indicated for lowered efficiency of whole-cell and acellular pertussis vaccines in elimination of mutated strains of B. pertussis from animal lung tissue in comparison with strains currently used for vaccine production.

[Application of DNA microcarriers for evaluation of mice splenocytes genes expression after immunization with pertussis vaccines and challenged with different B. pertussis strains]. / Zastosowanie mikronosników DNA do oceny profilu ekspresji genów komórek hodowli splenocytów myszy immunizowanych szczepionkami przeciwkrztuscowymi i zakazanych szczepami B. pertussis.

The study aimed at identification of splenocytes genes expression induced after immunization with whole-cell or acellular pertussis vaccines and subsequent challenge with B. pertussis strains of different ptxS1/prn genes alleles in correlation with differences in bacteria elimination from the lung tissue. Investigations were performed using DNA microcarriers. Analysis of genes expression generated through B. pertussis challenge on DNA microcarriers confirmed the role of specific B. pertussis virulence factors in differences of induced transcription response in the host. Level of differences in splenocytes transcriptional response of mice immunized with whole-cell pertussis vaccine was higher than that found in animals immunized with acellular pertussis vaccine. Level of genes expression change induced with B. pertussis 2316 strain was as much as twice higher than that found after challenge with B. pertussis 629 strain. Levels of gene expression change during challenge with B. pertussis 629 and 2316 strains were similar after immunization with acellular pertussis vaccine. Differences of transcriptional response induced with B. pertussis 2316 strain were associated with induction of immunological response, activation of inflammatory mediators, as well as with synthesis of heat shock proteins. Further studies performed with DNA microcarriers on sufficient number of B. pertussis strains might confirm this initial screening of genes identification involved in activation and repression of basic regulatory pathways after challenge with B. pertussis strains variant's for statistical evaluation of functional differences detected in presented study.

[Analysis of chosen parameters of immuno response in mice immunized with whole-cell or acellular pertussis vaccines and challenged with B. pertussis strains harbouring different ptxS1/prn allele genes combinations]. / Analiza wybranych parametrów immmunologiczej odpowiedzi myszy szczepionych pelno- lub bezkomórkowa szczepionka przeciw krztuscowi i zakazanych szczepami B. pertussis o róznych kombinacjach alleli genów ptxS1/prn.

Studies concerned evaluation of differences between parameters of cell-mediated immunity in mice, induced with whole-cell and acellular pertussis vaccines with subsequent challenge with B. pertussis strains harbouring different ptxS1/prn allele genes. In the study, concentrations of IFN-gamma/Il-2 and 1l-4/Il-5 in supernatants of cultured mice splenocytes have been determined to evaluate differences in Th1 or Th2 lymphocytes subpopulation response. Simultaneously, studies of intracellular expression of genes encoding of Il-2, Il-12, IFN-gamma and Il-4, Il-5, Il-10, Il-13 in mice splenocytes, and genes encoding factors involved in inflammatory process in the lung tissue (GM-CSF, TNF-alpha, Il-1beta, Il-6 i TGF-beta) have been performed on RNA level. The obtained results, confirmed high polarization of immunological response toward Th1 in mice immunized with DTP vaccine with whole-cell pertussis component, and toward Th2 in mice immunized with acellular pertussis vaccine. Inflammatory process in the lung tissue was more pronounced in animals immunized with whole-cell pertussis vaccine. There were no quantitative differences of analysed factors involved in the immune response among mice challenged B. pertussis strains containing different ptxS1/prn composition.

Bordetella pertussis strains circulating in Europe in 1999 to 2004 as determined by pulsed-field gel electrophoresis.

Clinical isolates of Bordetella pertussis collected during the year 2004 (n = 153) in eight European countries, Denmark, Finland, France, Germany, The Netherlands, Poland, Sweden, and United Kingdom, were analyzed by pulsed-field gel electrophoresis (PFGE), and their PFGE profiles were compared with those of isolates collected in 1999 (n = 102). The 255 isolates produced 59 distinct PFGE profiles. Among the 153 isolates from 2004, 36 profiles were found, while within the 102 isolates from 1999, 33 profiles were detected. One PFGE profile, BpSR11, was dominant (30% to 50%) in all countries except Denmark (10%) and Poland (0%). In comparison with 1999, there was an increase in BpSR11 prevalence in Finland in 2004 from 5% to 40%, coinciding with a major incidence peak. Some other PFGE profiles seemed to be associated with limited dissemination. Poland was the only country in which the most common actual European PFGE profiles were not found. In a dendrogram analysis, all common PFGE profiles were identified within PFGE group IV, and BpSR11 clustered together with PFGE subgroup IVbeta. Compared to the 1999 isolates, PFGE group V representative for pertactin variant prn3 strains had disappeared, and a new cluster was seen. In conclusion, some PFGE profiles, such as BpSR11, evidently have a higher capacity to spread, suggesting increased fitness to the present immunological environment. It is therefore of major interest to continue with surveillance programs of B. pertussis isolates, as both waning vaccine-derived immunity and strain variation may play a role in the persistence of pertussis.

[Genetic diversity analysis of isolates belonging to Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica species]. / Zastosowanie metody aflp do oceny genetycznego podobienistwa szczepow Bordetella pertussis, Bordetella parapertussis oraz Bordetella bronchiseptica.

In this study, Amplified Fragment Length Polymorphism (AFLP) method was used to track differences among human and animal isolates of B. pertussis, B. parapertussis and B. bronchiseptica species. One hundred and sixty representative strains of these species orginated from international and Polish bacterial collections were genotyped according to AFLP involving EcoRI/Msel and SpeI/ApaI restriction/ligation/amplification procedures. This study has confirmed high potential AFLP SpeI/ApaI procedure for intra-species differentiation of B. pertussis and B. bronchiseptica strains. Both AFLP EcoRI/MseI and SpeI/ApaI procedures have been found to be useful for species-specific classification in case of B. pertussis strains. In case of B. bronchiseptica or B. parapertussis species-specific classification, SpeI/ApaI procedure has been found more precise than EcoRI/MseI one.

[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.

[Genetic diversity analysis of isolates belonging to Bordetella genus]. / Analiza metoda RAPD genetycznej zmiennosci szczepów z rodzaju Bordetella.

In this study, RandomAmplified Polymorphic DNA (RAPD) method was used to track differences among human and animals isolates of eight known species of Bordetella genus. One hundred representative strains of these species from international and Polish bacterial collections were genotyped according to RAPD protocols using primer 1254 or 1247. Problems with reproducibility and discriminatory power, frequently discussed in literature, have been overcome by precise optimization procedure, which allowed to achieve reliable conditions for Bordetella species analysed. This study proved high potential of RAPD method using primers 1247/1254 for intra-species differentiation of B. bronchiseptica and B. hinzii isolates. Additionally RAPD method has been found to be useful for confirming B. avium and B. holmesii identification.

Potency of pertussis component in the DTP vaccine--an overview of three decade study in Poland.

In Poland, similar to many highly immunized Western countries, a recent increase in cases of pertussis has been observed. This study aims to evaluate the level of potency fluctuations of the pertussis component of Polish-produced DTP vaccine due to the changes having occurred in production and potency testing procedures. We compared the potency of the pertussis component of DTP vaccine lots produced and evaluated in similar periods and with similar production and testing procedures. Records of Kendrick test results performed over a 30-year period were available for analysis. This study confirms the role of different manufacturers, changes in vaccine strain compositions, in-house reference preparations used as reference vaccines in the Kendrick tests, and in mice of single strain sources in the potency values obtained. In addition, the comparisons performed revealed a downtrend in potency levels since 1992. Potency decrease in vaccine lots produced during 1992-1997 has been positively correlated to the lowering of the number of IOU/dose. Strain compositions of the DTP vaccine pertussis component and in-house references have been found to be associated with the fluctuation in potency estimations, and confirmed their crucial role in ensuring vaccine efficacy. Our study reveals that relative efficacy of the DTP vaccine produced in 1992-1997 might be lower than that of vaccines produced in other periods. This might in turn explain the increase in pertussis cases among children aged 5-15 years which is presently being observed in Poland.

[Laboratory assessment problems of the efficacy of acellular pertussis vaccines]. / Problemy laboratoryjnej oceny skutecznosci bezkomórkowych szczepionek przeciw krztuscowi.

Although composition of acellular pertussis vaccines is better defined than whole-cell vaccines, differences in the formulation, content, and detoxification of pertussis vaccine antigens led to a unique character of each of differently produced acellular vaccine. Currently used methods for laboratory evaluation of whole-cell pertussis vaccine efficacy were found not suitable for acellular vaccines. There is a strong need to perform analysis and evaluation of the safety and efficacy profiles of acellular pertussis vaccines combined with other vaccine antigens (e.g. Hib) both before and after conjugation. Mechanisms of interactions seen after conjugation inducing weaker immunogenicity or efficacy are still poorly recognized.

Sequence variation in pertussis S1 subunit toxin and pertussis genes in Bordetella pertussis strains used for the whole-cell pertussis vaccine produced in Poland since 1960: efficiency of the DTwP vaccine-induced immunity against currently circulating B. pertussis isolates.

The present study indicates that the appearance of the B. pertussis harbouring prn2 gene allele variant (not found among clinical isolates before 1990s) may have been induced by long-term vaccination in Poland with DTP-composed vaccine strains presenting exclusively prn1. However, ptxS1A allele of pertussis toxin subunit S1 encoding gene, predominant in the currently isolated B. pertussis strains, has been found in vaccine strains used for whole-cell pertussis component (wP) production of DTP vaccine in 1960-1978. This outrules the possibility that the appearance of ptxSIA allele might be related to vaccine pressure driven by non-ptxS1A vaccine strains used for long-term immunization with wP. Intranasal challenge animal model testing the efficiency of the clearance of B. pertussis strains harbouring different ptxS1/prn allele gene combinations revealed that currently produced DTwP vaccine may not contain adequate B. pertussis vaccine strains, since isolates with gene variants different from those observed in vaccine strains were eliminated from the lungs of the immunized animals with lower efficiency.

Pertussis in Poland.

BACKGROUND: Since 1997, an unexpected 2-5-fold increase in the incidence of pertussis has been reported in Poland in comparison with the previous 10 years, although the introduction of the diphtheria-tetanus-pertussis (DTP) vaccination in 1960 reduced the incidence of pertussis approximately 100-fold in the 1980s. The aim of the study was to analyse all available data on pertussis in Poland to identify the risks associated with its re-emergence. METHODS: Available data on notification, incidence, mortality, hospitalization, geographical distribution, incidence according to age, and diagnosis of pertussis were collected from national surveillance monographs and statistically evaluated. RESULTS: Analyses performed in the study found two periods of rising and falling trends: in the incidence before and after 1989, respectively. Moreover, after 1989, the age-specific incidence among children aged 0-4 years decreased, and among 5-9, 10-14, and 15-19 year olds increased in comparison to the previous decade. The incidence rate of pertussis among infants was similar in both decades analysed. Clustering of pertussis incidence increase in provinces along a line from North East to South West was observed. CONCLUSIONS: As vaccination coverage did not decrease and diagnostics have not been improved since the 1980s, it is possible that waning immunity and the appearance of Bordetella pertussis vaccine escape mutants are involved in the changing pertussis epidemiological parameters. Further monitoring studies, together with improving diagnostics, might allow more precise epidemiological data to be obtained. An additional booster dose of acellular pertussis vaccine at age 6 years has been included in the current vaccination schedule.

[Increase in the incidence of pertussis and quality of whole-cell pertussis component of the DTP vaccine produced in Poland. Part I. Potency of DTP vaccine]. / Wzrost zachorowan na krztusiec a wyniki analizy jakosci komponentu krztuscowego polskiej szczepionki DTP. Czesc I: Moc szczepionki DTP.

This study aimed to identify and to evaluate the level of potency fluctuations of the pertussis component of Polish-produced DTP vaccine lots produced within 1972-2001 due to the changes having occurred in production and potency testing procedures. The study confirms that higher potency values were obtained for vaccine lots produced since 80-ties, e.g. after changes of: references lots (1975), vaccine strains (1978) and source of animals used in Kendrick tests (1979). Additionally, the comparisons performed revealed a down trend in potency levels within 1992-1999 correlating to the lowering of the number of IOU/dose.

[Increase in the incidence of pertussis and quality of whole-cell pertussis component of the DTP vaccine produced in Poland. Part II. Consistency of control and production]. / Wzrost zachorowan na krztusiec a wyniki analizy jakosci komponentu krztuscowego polskiej szczepionki DTP. Czesc II: Jednorodnosc badan i produkcji.

This study aimed to identify and to evaluate the level of potency fluctuations of the pertussis component of Polish-produced DTP vaccine lots produced within 1972-2001 due to the changes having occurred in production and potency testing procedures. Obtained results showed high consistency of the production and control tests confirming relevant quality of country-produced DTP vaccine.