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1.
Med Dosw Mikrobiol ; 66(2): 89-98, 2014.
Article in Polish | MEDLINE | ID: mdl-25369655

ABSTRACT

INTRODUCTION: Highly pathogenic Y. enterocolitica bioserotype 1B/O8 is considered to be an important etiological agent of yersiniosis in Poland. Infections caused by Y. enterocolitica 1B/O8 became an important public health problem in Poland, especially because of their high potential of virulence and the unknown source of the bacteria. Y. enterocolitica 1B/O8 isolates recovered in Poland are genetically highly related and constitute single epidemic sensu stricto strain. The aim of the present study was to develop a time- and money-effective molecular assay for rapid identification of pathogenic Y. enterocolitica 1B/O8 isolates belonging to the epidemic strain. METHODS: In the first stage we performed a multiplex-PCR for four genetic markers: ail, ystA, irp1 and 16S rDNA sequence. In the next stage we designed a duplex-PCR-RFLP assay with BtsI endonuclease to detect/identify specific variant of an ysrR gene that is characteristic for epidemic strain of Y. enterocolitica 1B/O8 strain. The assay was tested against a panel of a consisted of a variety Yersinia enterocolitica and Y. pseudotuberculosis strains. RESULTS: All the tested Y. enterocolitica 1B/O8 strains were positive for all the genetic markers in multiplex-PCR assay what distinguished them from other tested Yersinia strains. In duplex-PCR-RFLP test all tested isolates of the epidemic strain were negative for ysrR digestion with BtsI endonuclease, while all tested reference strains of Y. enterocolitica 1B/O8 were positive. CONCLUSIONS: The assay developed in this study was two-stage/two-step molecular test efficiently distinguishing wild-type and the epidemic Y. enterocolitica 1B/O8 strain. Such test can be a useful screening tool for clinical, veterinary and food diagnostics, as well as for the purposes of epidemiological investigation.


Subject(s)
Polymerase Chain Reaction/methods , Yersinia Infections/microbiology , Yersinia enterocolitica/classification , Yersinia enterocolitica/isolation & purification , Disease Outbreaks , Genetic Markers/genetics , Humans , Poland/epidemiology , Polymorphism, Restriction Fragment Length , Serotyping , Species Specificity , Virulence/genetics , Yersinia Infections/epidemiology , Yersinia enterocolitica/genetics , Yersinia enterocolitica/pathogenicity
2.
Postepy Hig Med Dosw (Online) ; 68: 48-56, 2014 Jan 22.
Article in Polish | MEDLINE | ID: mdl-24491895

ABSTRACT

Campylobacter jejuni and Campylobacter coli are Gram-negative, microaerophilic bacteria which are worldwide in distribution, causing a zoonotic disease in humans called campylobacteriosis. These infections are mainly caused by eating contaminated food products, most often improperly prepared poultry meat. Campylobacteriosis usually takes the form of gastroenteritis, or inflammation of the intestines, and the characteristic symptoms are watery-mucous diarrhea often with the presence of blood in stool, nausea, vomiting, abdominal pain and fever. The epidemiological data suggest that in Europe, as well as in North America, bacteria of the genus Campylobacter, especially C. jejuni and C. coli, are the most commonly isolated pathogens in infections of the gastrointestinal tract in humans. Epidemiological data indicate that these organisms are a much more common cause of acute diarrhea, mostly in young children, than Salmonella and Yersinia. The lack of specific symptoms makes the diagnosis of campylobacteriosis necessary to carry out specialized microbiological diagnostics. Because so far these studies are performed in our country only in a few laboratories, the overwhelming number of cases of campylobacteriosis are not recorded in Polish epidemiological statistics. The purpose of this paper is to discuss issues related to the microbiological diagnosis of infections caused by C. jejuni and C. coli. It also describes the basic epidemiological and clinical data, as well as current treatment of campylobacteriosis.


Subject(s)
Campylobacter Infections/diagnosis , Campylobacter Infections/microbiology , Campylobacter coli/isolation & purification , Campylobacter jejuni/isolation & purification , Gastroenteritis/diagnosis , Gastroenteritis/microbiology , Campylobacter Infections/epidemiology , Causality , Comorbidity , Diarrhea/epidemiology , Diarrhea/microbiology , Feces/microbiology , Fever/microbiology , Gastritis/microbiology , Gastroenteritis/epidemiology , Humans , Inflammation/diagnosis , Inflammation/microbiology , Meat/microbiology , Salmonella
3.
BMC Infect Dis ; 13: 551, 2013 Nov 19.
Article in English | MEDLINE | ID: mdl-24252165

ABSTRACT

BACKGROUND: Recommendations for diphtheria immunization are to apply an effective primary immunization in infancy and to maintain immunity throughout life. Immunity against diphtheria depends primarily on antibody to the diphtheria toxin. This study evaluated the seroprevalence of IgG diphtheria antitoxin in sera of healthy children, adolescents and adults in Poland. METHODS: A total of 1387 serum samples collected between 2010 and 2012 from individuals with ages ranging from 1 month to 85 years were investigated. Antibody concentrations were measured with an enzyme-linked immunosorbent assay (Anti-Diphtheria Toxoid ELISA IgG, Euroimmun, Germany). RESULTS: The results showed that among 1387 individuals examined, 547 (39.4%) had anti-diphtheria toxoid IgG antibody levels below 0.1 IU/ml (36.9% ≤ 18 years and 40.5% >18 years old, respectively). The 212 (50.8%) children and 542 (55.9%) adults showed only basic protection (0.1-1.0 IU/ml) and need immediate booster. High levels of anti-diphtheria toxoid IgG antibodies (>1.0 IU/ml) were found more often in children and adolescent (12.2%) than in adults (3.6%) and this was statistically significant (P < 0.05). The proportion of seronegatives (< 0.1 IU/ml) in children below 2 years old, adolescents and young adults to 25 years old decreased from 53.5% to 17.4%. However, in older individuals the seronegative proportion tended to increase with age, from 22.7% in adults (26-30 years old) to 67.1% in subjects > 60 years old. Characteristically, in individuals > 40 years old high levels of anti-diphtheria toxoid IgG antibodies (>1.0 IU/ml) were not seen. There were no statistically significant differences in results in relation to gender. CONCLUSIONS: The present study showed inadequate immunity levels to diphtheria amongst the Polish population, especially in adults > 40 years old and children ≤ 2 years old. To prevent reemergence of diphtheria an information campaign reminding people about recommendations concerning diphtheria booster vaccination in adults should be conducted. Moreover, the immunogenicity of the DTP vaccine used in Poland should be verified.


Subject(s)
Antibodies, Bacterial/blood , Diphtheria Toxoid/immunology , Diphtheria/epidemiology , Diphtheria/immunology , Immunoglobulin G/blood , Adolescent , Adult , Child , Corynebacterium diphtheriae/immunology , Diphtheria/prevention & control , Diphtheria-Tetanus-Pertussis Vaccine/immunology , Female , Humans , Infant , Male , Poland/epidemiology , Seroepidemiologic Studies , Young Adult
4.
Med Dosw Mikrobiol ; 64(1): 79-85, 2012.
Article in Polish | MEDLINE | ID: mdl-22808733

ABSTRACT

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.


Subject(s)
Antigens, Bacterial/blood , Freezing , Immunoglobulin A/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Tissue Preservation/methods , Tissue Preservation/statistics & numerical data , Enzyme-Linked Immunosorbent Assay , Humans
5.
Med Dosw Mikrobiol ; 64(4): 277-83, 2012.
Article in Polish | MEDLINE | ID: mdl-23484419

ABSTRACT

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.


Subject(s)
Bordetella Infections/epidemiology , Bordetella Infections/microbiology , Bordetella parapertussis/immunology , Cough/epidemiology , Immunoglobulin Isotypes/blood , Lipopolysaccharides/immunology , Adolescent , Adult , Bordetella Infections/diagnosis , Bordetella parapertussis/isolation & purification , Causality , Child , Child, Preschool , Comorbidity , Cough/immunology , Female , Humans , Immunoglobulin A/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Incidence , Male , Middle Aged , Poland/epidemiology , Seroepidemiologic Studies , Sex Distribution , Sex Factors , Young Adult
6.
Med Dosw Mikrobiol ; 63(1): 65-71, 2011.
Article in Polish | MEDLINE | ID: mdl-22184899

ABSTRACT

Salmonella Typhimurium and Salmonella Enteritidis are the two predominant serogroups, responsible for about 80% of all human cases of salmonelosis in Poland. Therefore we compared the usefulness of lipopolysaccharides antigens extracted by phenol (Westphal method) and trichloroacetic acid (Boivine method) from Salmonella Typhimurium and Enteritidis in ELISA method for the determination of antibodies. We used one home - made LPS antigen and two others commercially available antigens from SIGMA - Aldrich. Our study showed that the presence of antibodies was found in 35 (74.5%) sera from 47 samples from patients with suspected salmonelosis. There was no significant statistical differences of frequency of appearance of antibodies to all three Salmonella antigens in sera from patients with salmonelosis and in sera from control group. This study showed that all three antigens are useful for determination of IgA, IgG, IgM antibodies for Salmonella serogroup B and D in routine serological diagnosis of salmonelosis. However, it should be considered possibility of cross-reaction between LPS antigen of Salmonella and antibodies to Yersinia enterocolitica which could be correlated with similarity between somatic antigens of these two pathogens.


Subject(s)
Antigens, Bacterial/isolation & purification , Lipopolysaccharides/isolation & purification , Salmonella Infections/diagnosis , Salmonella Infections/microbiology , Salmonella enteritidis/chemistry , Salmonella typhimurium/chemistry , Serologic Tests/methods , Enzyme-Linked Immunosorbent Assay , Humans , Lipopolysaccharides/chemistry , Phenol/chemistry , Salmonella enteritidis/immunology , Salmonella typhimurium/immunology , Serotyping , Trichloroacetic Acid/chemistry
7.
Med Dosw Mikrobiol ; 63(3): 255-61, 2011.
Article in Polish | MEDLINE | ID: mdl-22184920

ABSTRACT

Diagnosis of previous C. jejuni infections in GBS patients are mostly based on serological findings. However, there are not consensus what kind of antigen should be used in the serological assays. In our study we used ELISA with four different antigen preparations for investigation of specific antibodies to C. jejuni in serum samples obtained from 6 children with GBS. In all patients the high level of IgA and IgG antibodies to LPS were diagnosed. The antibodies in particular classes of immunoglobulins to recombinant proteins (Mikrogen), termostabile antigen and whole-cell antigen (Virion/Serion) of C. jejuni were diagnosed only in some of the children with GBS. However, in comparison to the control groups, the ELISA with recombinant proteins was most specific. Moreover, in two of the patients a characteristic decline of the level of antibodies to recombinant proteins in sera obtained in acute and chronic phase of disease have been observed. The results of this study showed that serodiagnosis, specially based on recombinant antigens, may be a reliable marker of recent or previous infection caused by C. jejuni in patients with GBS.


Subject(s)
Antibodies, Bacterial/blood , Campylobacter Infections/immunology , Campylobacter jejuni/immunology , Guillain-Barre Syndrome/complications , Child , Child, Preschool , Cyclohexanecarboxylic Acids/blood , Guillain-Barre Syndrome/immunology , Humans , Immunoglobulin G/blood , Serologic Tests
8.
Med Dosw Mikrobiol ; 63(4): 299-304, 2011.
Article in Polish | MEDLINE | ID: mdl-22384662

ABSTRACT

The aim of the study was to investigate the presence of antibodies to lipopolysaccharides obtained by modified Boivin's method from E. coli serotype O104:H4 and O26, O103, O111, O121, O145, O157 in sera of 7 patients with acute diarrhea, suspected in clinical investigation for infection caused by E. coli O104:H4. Additionally, to determine the cut-off levels, the 75 sera from blood donors were tested. The high level of antibodies to LPS E. coli O104 was diagnosed in three patients from family outbreak caused by E. coli serotype O104:H4. In one of those patients, 7-years boy with HUS, we observed also a significant decrease of level of IgA, IgG and IgM antibodies in serum sample obtained in chronic phase of the disease. Furthermore, we showed that two others patients with clinical evidence of VTEC infection, not connected with this family outbreak, had a high level of antibodies to E. coli of other serotypes: one to O157 and one to O103. We did not observe presence of antibodies to LPS from E. coli O26, O111, O121 i O145 in the sera of tested patients. In conclusion, we confirmed that ELISA based on lipopolysaccharides obtained from different serotypes of E. coli may be helpful in laboratory diagnosis of infection caused by VTEC in humans.


Subject(s)
Antibodies, Bacterial/blood , Enzyme-Linked Immunosorbent Assay , Escherichia coli Infections/diagnosis , Escherichia coli Infections/immunology , Lipopolysaccharides/immunology , Shiga-Toxigenic Escherichia coli/immunology , Adult , Aged , Child , Female , Humans , Male , Poland , Serotyping , Shiga-Toxigenic Escherichia coli/classification
9.
Med Dosw Mikrobiol ; 63(4): 315-20, 2011.
Article in Polish | MEDLINE | ID: mdl-22384664

ABSTRACT

The Binax and the Biotest urinary antigen kits for detection of L. pneumophila antigen were compared by testing of selected 67 urine samples obtained from EWGLI as reference samples in External Quality Assessment Scheme. Thirty nine were positive with the Binax kit (100% of sensitivity), and 33 were positive with the Biotest (84.6% of sensitivity). The test specificities were 100% for the both kits. It was concluded that the Binax kit was more suitable for the routine diagnosis of Legionella infections than the Biotest kit.


Subject(s)
Antigens, Bacterial/urine , Immunoenzyme Techniques , Legionella pneumophila/immunology , Legionella pneumophila/isolation & purification , Humans , Legionella pneumophila/classification , Legionellosis/diagnosis , Legionellosis/urine , Serotyping
10.
Med Dosw Mikrobiol ; 62(2): 109-17, 2010.
Article in Polish | MEDLINE | ID: mdl-20873483

ABSTRACT

We compared the 2.653 results of routine serological investigations performed in three different laboratories in Poland for the diagnosis of pertussis. One of the laboratories used the NovaLisa Bordetella pertussis kit produced by NovaTec GmbH and the two others used Bordetella pertussis ELISA kit produced by Genzyme Virotech GmbH. In first laboratory the diagnostic level of IgA antibodies to the pertussis toxin was observed in 11.0%, IgG in 52.7% and IgM in 27.4% serum samples. In the second and third laboratory the diagnostic level of IgA antibodies were found respectively in 22.2% and 40.1%, IgG in 46.8% and 66.4% and IgM only in 8.7% and 4.8% serum samples. In total, IgA antibodies were found in 28.0%, IgG antibodies in 56.0% and IgM antibodies in 14.3% serum samples obtained from patients suspected in clinical investigation for pertussis. We have observed that the frequency of detection of the antibodies in children and adolescents increased with age reaching its peak among individuals with age range between 11-20 years. We also found statistical significant higher frequency of IgA, IgG and IgM antibodies to B. pertussis in outpatients than in hospital patients.


Subject(s)
Serologic Tests/methods , Whooping Cough/diagnosis , Adolescent , Antibodies/immunology , Child , Female , Growth and Development/immunology , Humans , Male , Poland , Reagent Kits, Diagnostic/statistics & numerical data , Whooping Cough/immunology , Young Adult
11.
Med Dosw Mikrobiol ; 62(1): 21-8, 2010.
Article in Polish | MEDLINE | ID: mdl-20564967

ABSTRACT

The ELISA were performed on polystyrene microtiter plates (Nunc, MaxiSorp) coated with LPS (2a antigen) at the final concentration of 10 microg/ml. The antigen was extracted from Klebsiella rhinoscleromatis Rh32 by the trichloroacetic acid and separated by ethanol (Boivin method). The antibodies against the LPS were detected by ELISA in serum samples collected from 65 patients suspected in clinical investigation for rhinoscleroma in Poland from 1970 to 2009. Additionally, the specificity of the antigen was tested using serum sample of immunized rabbit and 30 sera of patients from control group, with high level of antibodies to different bacterial pathogens. All serum samples were diluted 1:100. The concentrations of IgA, IgG and IgM antibodies were expressed as optical density (OD) measured at the wavelength of 450 nm. The cut-off limit of serum antibodies was set at mean antibody OD determined in the sera of 30 blood donors exceeded by three standard deviations. The presence of IgA and IgG antibodies were detected by ELISA in 33 (50,8%) and IgM in 28 (43,1%) of patients. Most of the serum samples (75%) with high level of specific antibodies were obtained from patients before 1980. On the other hand antibodies to K. rhinoscleromatis were detected only in 2 (6,7%) patients from the control group and none of blood donors. In conclusion, our home-made ELISA, based on purified LPS of K. rhinoscleromatis showed high specificity and sensitivity in the diagnosis of antibodies to K. rhinoscleromatis in comparison to the complement fixation test. The presence of high level of specific IgA, IgG and IgM antibodies in the sera obtained in different stages of disease may showed that during the rhinoscleroma is permanent stimulation of antibody production.


Subject(s)
Antibodies, Bacterial/analysis , Klebsiella pneumoniae/immunology , Klebsiella pneumoniae/isolation & purification , Rhinoscleroma/diagnosis , Rhinoscleroma/microbiology , Animals , Enzyme-Linked Immunosorbent Assay , Humans , Immunoglobulin A/analysis , Immunoglobulin G/analysis , Lipopolysaccharides/immunology , Rabbits , Rhinoscleroma/immunology , Sensitivity and Specificity
12.
Pneumonol Alergol Pol ; 78(1): 54-9, 2010.
Article in Polish | MEDLINE | ID: mdl-20162519

ABSTRACT

Legionella pneumophila is an important causative agent of pneumonia in humans which is difficult to diagnose because the signs and symptoms are nonspecific and do not distinguish Legionella infection from other common causes of pneumonia. Currently, the diagnosis of Legionnaires' disease is based on phenotyping (culture, antibody detection in human sera, antigen detection in urine) and genotyping methods such as PCR (polymerase chain reaction). This review focuses on current diagnostic tests for surveillance of Legionella pneumophila infections in Poland.


Subject(s)
Legionella pneumophila/isolation & purification , Legionnaires' Disease/diagnosis , Legionnaires' Disease/microbiology , Antigens, Bacterial/blood , Antigens, Bacterial/urine , Bacteriological Techniques/methods , DNA, Bacterial/analysis , Humans , Immunoenzyme Techniques , Legionnaires' Disease/blood , Legionnaires' Disease/urine , Poland , Polymerase Chain Reaction/methods , Practice Guidelines as Topic , Sensitivity and Specificity
13.
Med Dosw Mikrobiol ; 61(2): 175-82, 2009.
Article in Polish | MEDLINE | ID: mdl-19780496

ABSTRACT

A panel of 33 different antigens, among them lipoproteins, glicolipids and proteins, of Mycoplasma pneumoniae used in commercial western-blotting (Virotech) were assessed for reactivity with sera of patients with mycoplasmosis and other bacterial infections of variable etiology. In addition, commercial ELISA (Virotech) with recombinant proteins as antigen and complement fixation test (CFT) with in-house prepared glicolipid-protein antigen were also assessed for comparison. The proteins with molecular weight of 170 kDa (P1) and 90 kDa (P90) were most recognized by the serum samples of patients with mycoplasmosis. The reactions of proteins P50, P47 and Fts monomer with positive sera were not such often and the response was usually weak. The other proteins of M. pneumoniae, particularly P88, Repet. Prot. or P20, were recognized occasionally or at all. We observed also the often reactions ofglicolipids of M. pneumoniae with IgM antibodies. The result of the study showed that the commercial ELISA (Virotech) was equivalent in sensitivity and specificity to the CFT in the case of sera obtained in the acute phase of mycoplasmosis (90.7% of agreement of results in the class IgA, 85.6% in the class IgG and 100% in the class IgM). Analytical specificity studied by screening serum samples from patients with different bacterial infections and blood donors have shown lower specificity of ELISA in compared to western-blotting. The present study confirmed the earlier observations of the high usefulness of P1 protein and P90 protein for reliable serologic diagnosis of acute M. pneumoniae infection.


Subject(s)
Antigens, Bacterial/analysis , Mycoplasma pneumoniae/immunology , Mycoplasma pneumoniae/isolation & purification , Pneumonia, Mycoplasma/diagnosis , Serologic Tests/methods , Antigens, Bacterial/chemistry , Blotting, Western , Enzyme-Linked Immunosorbent Assay , Humans , Molecular Weight
14.
Przegl Epidemiol ; 63(4): 513-8, 2009.
Article in Polish | MEDLINE | ID: mdl-20120949

ABSTRACT

Five hundred of results of commercial ELISA test (NovaLisa Bordetella pertussis, NovaTec) used in routine diagnosis of IgA, IgG and IgM antibodies to pertussis toxin in serum samples obtained from patients with respiratory tract infection were analyzed. The diagnostic level of IgA antibodies was observed in 54 (10.8%), IgG in 258 (51.6%) and IgM in 136 (27.2%) serum samples. The frequency of detection of the antibodies in children and adolescents increased with age reaching its peak among individuals with age range between 16-20 years. The results of our study showed also a high frequency of IgG antibodies to the pertussis toxin in the adults with respiratory tract infections. We also found statistical significant higher frequency of IgM antibodies, but not IgA and IgG, to pertussis toxin in women than in men.


Subject(s)
Antibodies, Bacterial/blood , Immunoglobulin A/blood , Immunoglobulin G/blood , Immunoglobulin M/blood , Pertussis Toxin/immunology , Respiratory Tract Infections/immunology , Adolescent , Adult , Age Distribution , Child , Enzyme-Linked Immunosorbent Assay , Female , Humans , Male , Middle Aged , Pertussis Vaccine/administration & dosage , Poland/epidemiology , Respiratory Tract Infections/blood , Respiratory Tract Infections/epidemiology , Respiratory Tract Infections/prevention & control , Sex Distribution , Young Adult
15.
Med Dosw Mikrobiol ; 61(4): 375-9, 2009.
Article in Polish | MEDLINE | ID: mdl-20201328

ABSTRACT

The prevalence of human IgG1-IgG4 to sonicated antigen of M. pneumoniae in the 138 sera obtained from patients with mycoplasmosis was analysed. Antibodies of IgG1 were diagnosed in 47 (34.1%), IgG2 in 42 (30.4%) and IgG3 subclass in 58 (42.0%) serum samples. The concentration of IgG4 was below detection level. Generally, the frequency of occurrence of IgG2 antibodies increased with age reaching its peak among adults. On the other hand IgG1 and IgG3 for M. pneumoniae were diagnosed more often in serum samples obtained from children than from adults. We did not find any essential changes in the pattern of IgG subclass during the course of infection however it seems that the level of IgG3 antibodies decreased faster than IgG1 which may be caused by the fact that the IgG3 antibodies have a much shorter half-life in comparison to the IgG1.


Subject(s)
Aging/immunology , Immunoglobulin G/blood , Mycoplasma pneumoniae/immunology , Pneumonia, Mycoplasma/immunology , Adolescent , Adult , Child , Child, Preschool , Half-Life , Humans , Immunity, Humoral/immunology , Infant , Infant, Newborn , Pneumonia, Mycoplasma/microbiology , Young Adult
16.
Med Dosw Mikrobiol ; 60(2): 121-9, 2008.
Article in Polish | MEDLINE | ID: mdl-18819448

ABSTRACT

Campylobacter jejuni and Campylobacter coli are the most common bacterial cause for acute diarrheal illnesses in developed countries. The aim of this study was to evaluate the antigenic properties of Campylobacterjejuni and Campylobacter coli proteins in western-blot assay. Whole-cell components of Campulobacter jejuni and Campylobacter coli were separated by sodium dodecyl sulfate-polyacrylamide gel electroforesis. Using this method we detected in all seven C. jejuni strains 21 peptides migrating between 180-29 kDa. All three Ccoli strains had a 17 bands migrating with the same molecular weight range. Proteins were transferred electrophoretically to nitrocellulose paper for immunoblotting experiments. The 74 kDa protein reacted strongly in all classes ofimmmunoglobulin with all tested human serum samples. We observed that this protein reacted also with human immunoglobulins for Salmonella and Yersinia sp. This cross-reaction observed for this protein could give false positive results in routine diagnosis of C. jejuni infections. The proteins with molecular weight of: 92, 62, 56, 52, 45-43, 29 kDa were most recognized in the 20 human serum samples. The other proteins of Cljejuni and C. coli, particularly in the 68-50 kDa and 45-31 kDa regions, were recognized occasionally and the response to these in reconvalescent sera was usually weak. The result of this study showed that the proteins with molecular weight: 92, 62, 56, 52, 45-43 and 29 kDa can be use in routine serological diagnostic of campylobacteriosis.


Subject(s)
Antibodies, Bacterial/immunology , Antigens, Bacterial/immunology , Bacterial Proteins/immunology , Campylobacter coli/immunology , Campylobacter jejuni/immunology , Antigens, Bacterial/isolation & purification , Bacterial Proteins/isolation & purification , Blotting, Western , Campylobacter Infections/diagnosis , Campylobacter Infections/immunology , Cross Reactions , Epitopes , Humans , Serologic Tests
17.
Med Dosw Mikrobiol ; 60(4): 289-95, 2008.
Article in Polish | MEDLINE | ID: mdl-19382600

ABSTRACT

The commercially available enzyme-linked immunosorbent assays (ELISA recomWell Campylobacter) from Mikrogen was evaluated for the diagnosis of Campylobacter jejuni and Campylobacter coli infections. Serum samples from 20 healthy controls, 44 persons with symptoms of primary Campylobacter infection and 24 serum samples from patients with Yersinia enterocolitica or Salmonella infections were tested. This ELISA assay detects IgA and IgG antibodies against three recombinant antigens of the Campylobacter jejuni and Campylobacter coli: OMP 18 (18 kDa), PEB4 (31 kDa) and P39 (39 kDa). The healthy controls showed significantly lower antibody titers in all two immunoglobulin classes. The IgA antibodies were diagnosed only in 2 (18.2%) serum samples obtained from patients with bacteriologically confirmed campylobacteriosis. The presence of IgG antibodies was confirmed in 82% of serum samples. Furthermore, we showed that 66.7% of the 33 serum samples obtained from the patients suspected for campylobacteriosis not confirmed by isolation, were positive for IgG and 15.2% for IgA antibodies. We observed also not specific reactions in ELISA recom Well Campylobacter with sera obtained form patients with yersiniosis and salmonelosis. This study demonstrates the usefulness of commercially available assay for the routine diagnosis of Campylobacter infection but with some limitations.


Subject(s)
Campylobacter Infections/diagnosis , Campylobacter coli/isolation & purification , Campylobacter jejuni/isolation & purification , Enzyme-Linked Immunosorbent Assay/instrumentation , Serologic Tests/instrumentation , Campylobacter Infections/immunology , Diagnosis, Differential , Humans , Immunoglobulin G/blood , Yersinia Infections/diagnosis , Yersinia enterocolitica
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