[Problems in the serological diagnosis of atypical pneumonia caused by Legionella pneumophila and Mycoplasma pneumoniae]. / Problemy w serologicznej diagnostyce atypowych zapalen pluc na przykladzie wyników badan w kierunku zakazen wywolywanych przez Legionella pneumophila oraz Mycoplasma pneumoniae.

INTRODUCTION: The clinical presentation of atypical pneumonia is often similar to the presentation of more typical bacterial pneumonias and the etiological agent must be confirmed by laboratory diagnosis. This article will discuss the problems in the serological diagnosis of atypical pneumonia caused by Legionella pneumophila and Mycoplasma pneumoniae which are the agents most commonly associated with atypical pneumonia. Specifically, seeking the possibility of non-specific response, we evaluated the prevalence of antibodies to M. pneumoniae in serum samples obtained from patients suspected in clinical investigation for legionellosis. METHODS: The total numbers of 261 serum obtained from patients suspected in clinical investigation for legionellosis, were tested by in-house ELISA with M. pneumoniae sonicated antigen. Some of the positive sera were also re-tested by western-blot with high specific recombinant M. pneumoniae P1 protein. RESULTS: The diagnostic significant level of IgA antibodies to M. pneumoniae were diagnosed by ELISA in 71 (27,2%) of tested serum samples. Some of the IgA-positive sera have also high level of IgG and IgM antibodies to M pneumoniae (respectively 4,2% and 6,5%). Most from the 18 selected positive results obtained by ELISA were also confirmed by western-blot. It was characteristic that IgA antibodies to M pneumoniae were detected more than three times often in serum samples with positive serological tests for Legionnaires' disease than in samples with negative results for L. pneumophila. CONCLUSIONS: This study showed the possibility of non-specific reactions in serological diagnosis of atypical pneumonia. However, according to the data of the literature, co-infections of L. pneumophila and M pneumoniae can not be excluded.

Seroprevalence of IgG antibodies against tetanus toxoid among different age groups in Poland.

INTRODUCTION AND OBJECTIVE: Vaccination is the most effective and reliable strategy for preventing the morbidity of tetanus. The aim of the study is to investigate the seroprevalence of antibodies to tetanus toxoid among healthy persons across all age groups to determine the level of vaccine-induced immunity in the population, and to identify which age group should be targeted for a booster dose. MATERIAL AND METHODS: A total of 2,842 serum samples collected between 2010 - 2019 from individuals aged from 1 month - 97 years were investigated. Anti-tetanus IgG antibody concentrations (IU/ml) were measured by an enzyme-linked immunosorbent assay. In addition, the avidity of antibodies was determined using an in-house ELISA. RESULTS: The results showed that among the 2,842 individuals, 147 (5.2%) had anti-tetanus toxoid IgG antibody levels below 0.1 IU/ml and another 1,519 (53.4%) subjects showed only basic protection (0.1-1.0 IU/ml) and needed immediate booster. High levels of anti-tetanus toxoid IgG antibodies (>1.0 IU/ml) were found more often in young adults at the age 21-40 years (55.5%, GMT=1.15). Importantly, these antibodies also had the highest avidity. With age, the percentage of high positives decreased, as well as the geometric mean and avidity of antibodies, reaching the lowest level in subjects over 70 years of age (13.3%; GMT=0.19). Characteristically, a higher percentage of high positive results was observed in men (42.6%) than in women (34.3%). CONCLUSIONS: The study showed adequate immunity levels to tetanus amongst the Polish population, especially in children, adolescents, and young adults. However, those from older age groups should receive booster doses of the vaccine.

[Serum immunoglobulin IgG subclass distribution of antibody responses to pertussis toxin and filamentous hemagglutinin of Bordetella pertussis in patients with whooping cough]. / Wystepowanie przeciwcial dla toksyny krztuscowej i wlókienkowej hemaglutyniny Bordetella pertussis w poszczególnych podklasach IgG w przebiegu krztusca.

INTRODUCTION: The present study was aimed at determining the IgG subclass distribution against pertussis toxin (PT) and filamentous hemagglutinin (FHA) of Bordetella pertussis in patients with whooping cough. METHODS: The total number of 222 serum samples obtained from patients suspected in clinical investigation for pertussis were tested separately by in-house ELISA for the presence of IgG antibodies to pertussis toxin and filamentous hemagglutinin. The percentage distribution of specific anti-PT and anti-FHA IgG subclass response was calculated only on the basis of group of sera confirmed in the present study as positive for total IgG antibodies (183 sera to PT antigen and 129 to FHA antigen). Paired serum specimens were obtained from 36 patients. Based on the results of determining the level of antibodies in the sera of 40 blood donors, the cut-off limit of serum antibodies for each subclass was set at arithmetic mean plus two standard deviations. RESULTS: Antibodies of IgG1 to pertussis toxin and filamentous hemagglutinin were diagnosed in 151 (82.5%) and 99 (76.7%), IgG2 in 72 (39.0%) and 50 (38.8%), IgG3 in 17 (9.3%) and 43 (33.3%), IgG4 in 55 (30.1%) and 53 (41.1%) serum samples, respectively. There were no significant differences in percentage of sera with IgG1, IgG2 and IgG3 in relation to age of the patients. However, the frequency of occurrence of IgG4 antibodies was highest in the group of the youngest children to the age of 6 years old (61.8% for PT and 68.0% for FHA), and decrease with age, reaching the minimum in the group of patients above 40 years old (13.2% and 4.2% for PT and FHA, respectively). We also found significantly higher frequency of IgG4 to PT and FHA antigens in men than in women. Statistically significant, essential changes in the pattern of IgG subclass during the course of infection were not found. CONCLUSIONS: In conclusion, this study showed that all four subclasses of IgG antibodies to pertussis toxin and filamentous hemagglutinin are produced during whooping cough.

[Characteristics of Clostridium tetani and laboratory diagnosis of tetanus]. / Charakterystyka Clostridium tetani i laboratoryjna diagnostyka tezca.

The causative agent of tetanus is the obligate anaerobic bacterium--Clostridium tetani. These bacteria form endospores that are able to survive long periods of exposure to air and other adverse environmental conditions. Infection generally occurs through wound contamination. We can distinguish several forms of tetanus: generalized, local and neonatal. Diagnosis of tetanus is based primarily on the patient's clinical symptoms (muscle cramps, painful back muscle spasms, generalized contractions of the arcuate curvature of the body) as well as on microbiological diagnosis. This article is a brief review of C. tetani and diagnosis of infections caused by these organisms in humans.

[Effect of multiple freeze-thaw cycles on detection of IgA, IgG and IgM antibodies to selected bacterial antigens]. / Wplyw wielokrotnego, cyklicznego zamrazania i rozmrazania próbki surowicy na poziom przeciwcial klasy IgA, IgG i IUgM dla wybranych antygenów bakteryjnych.

INTRODUCTION: In presented study we investigated the effect of multiple freeze-thaw cycles of human sera on the determination of IgA, IgG and IgM antibodies to selected bacterial antigens. METHODS: A panel of 15 serum samples with elevated levels of antibodies to Mycoplasma peumoniae, Yersinia enterocolitica and Salmonella spp. were used (5 positive sera for each pathogen). One set of aliquots designed as the baseline, was taken and stored at 4-8o C for the remainder for the study. The remaining seven sets of aliquots were divided into two parts and repeatedly frozen respectively at two different temperatures: -65 degrees C and -25 degrees C. Once a day the aliquot sets were removed from the freezer and allowed to stand at room temperature for approximately 1 h until completely thawed. For the determination of the level of antibodies the sera after: 2, 5, 10, 15, 20, 25 and 30 freeze/cycle were used. The measurement of IgA, IgG and IgM antibodies was done using a home-made ELISA with four different antigens: whole-cell antigen of M. pneumoniae FH strain, LPS and Yop antigens of Y. enterocolitica serotype O:3 and LPS extracted by Westphal method from Salmonella serogroup B +D. The results were presented as the arithmetic mean of the antibody titre in five sera which were treated by the same number of freeze-thaw cycles. RESULTS: There was no significant statistic difference between levels of antibodies in unfrozen and frozen sera even after 30 freeze-thaw cycles. Depending of the antigen used in ELISA a slight varations in the level of antibodies were observed but the changes were small and not clinically significant. Examination of the ELISA values does not suggest any consistent nonlinear trend in levels of IgA, IgG and IgM antibodies in sera frozen at -65 degrees C as well at -25 degrees C. CONCLUSIONS: Our study demonstrates that the IgA, IgG and IgM antibody activity levels measured for M. pneumoniae, Y enterocolitica and Salmonella antigens are stable even after 30 freeze-thaw cycles.

[Seroprevalence of antibodies against lipopolysaccharides of Bordetella parapertussis in patients with prolonged caught in Poland]. / Czestosc wystepowania przeciwcial dla lipopolisacharydów paleczek Bordetella parapertussis u osób z przewleklym kaszlem w Polsce.

INTRODUCTION: Bordetella parapertussis is a bacterium closely related to Bordetella pertussis, also causes a pertussis - like symptoms in humans. Because of unsatisfactory level of routine microbiological diagnosis of B. parapertussis infections in Poland most of parapertussis cases are not reported. The aim of the presented study was to investigate incidence of B. parapertussis in patients with cough in Poland using serology method. METHODS: Serum IgA, IgG and IgM antibodies were determined in 1192 serum samples obtained from patients with respiratory infections and chronic cough and who were previously suspected of B. pertussis infection. As a control we used 258 sera from healthy people - blood donors. The LPS antigen was extracted by Westphal method from wild B. parapertussis strain isolated in Poland. For exclusion of possible false positive results with B. pertussis some of the sera were tested against the purified pertussis toxin (PT). RESULTS: The diagnostically significant level of IgA antibodies to LPS of B. parapertussis was detected in 11.9%, IgG in 12.2% and IgM in 9.6% serum samples. More often the antibodies were diagnosed in women than in men. In 63 serum samples, previously positive in NovaTec ELISA with mixed antigen of pertussis toxin and filamentous hemagglutinin we found also IgA and IgG antibodies to LPS of B. parapertussis. However, after use of purified pertussis toxin antigen in ELISA we confirmed the B. pertussis infections only in 28 cases. CONCLUSIONS: This study shows that B. parapertussis is a serious causative agent of infections in patients with prolonged caught in Poland. The serodiagnosis ofparapertussis should be conducted with sera obtained from patients suspected in clinical investigation for pertussis but negative in serological investigation with purified pertussis toxin antigen.

Duration of protection against Bordetella pertussis infection elicited by whole-cell and acellular vaccine priming in Polish children and adolescents.

BACKGROUND: In the context of reported resurgence of pertussis in the last decade, researchers hypothesized that acellular (aP) pertussis vaccines elicit a shorter-lived protection compared to whole-cell (wP) pertussis vaccines. However, in the studies seeking to demonstrate this hypothesis, exposure to each vaccine type was not concurrent, and contradictory epidemiologic modeling questioned its validity. The context of pertussis vaccination history in Poland, with both vaccine types used concurrently in comparable proportions, provided an opportunity to investigate this hypothesis. We sought to compare waning of protection by primary series vaccine type by measuring anti-pertussis toxin antibody concentrations as proxy for recent infection. MATERIALS AND METHODS: Serological samples from 2,745 children and adolescents aged ≥5 years and <16 years and with completed 5-dose pertussis vaccination series were tested by ELISA for pertussis toxin (PT) antibodies. Participants were stratified by type of priming vaccine (wP or aP). Vaccination timeliness and priming-specific trends in anti-PT antibody levels by time since last vaccine dose were analyzed. RESULTS: A total of 1,161 (42.5%) children received wP vaccines, and 1,314 (48.1%) received aP vaccines for their primary series and toddler booster. Overall, 53.57% of the subjects received doses 2-4 in a timely manner, while only 41.52% received all 5 doses at the recommended intervals. Using GMCs or seropositivity measures, both priming groups showed a re-increase in anti-PT antibody levels signing infection in recent years from 8 years after the school-entry booster onward. Comparisons did not show any significant differences between the two groups in the timing or intensity of this re-increase. CONCLUSION: Our results clearly confirm that vaccine-elicited immunity against pertussis wanes among adolescents even after a complete infant, toddler and school-entry vaccination series. The timing and intensity of the waning of protection appear similar with whole-cell as with acellular pertussis vaccines.

Increasing role of arthropod bites in tularaemia transmission in Poland - case reports and diagnostic methods.

The study describes four cases of tularaemia - one developed after contact with rabbits and three developed after an arthropod bite. Due to non-specific clinical symptoms, accurate diagnosis of tularaemia may be difficult. The increasing contribution of the arthropod vectors in the transmission of the disease indicates that special effort should be made to apply sensitive and specific diagnostic methods for tularaemia, and to remind health-care workers about this route of Francisella tularensis infections. The advantages and disadvantages of various diagnostic methods - molecular, serological and microbiological culture - are discussed. The PCR as a rapid and proper diagnostic method for ulceroglandular tularaemia is presented.