[Does sealing the oval window in addition to the round window bring an advantage in reserve therapy of acute idiopathic deafness?] / Bringt die Abdeckung des ovalen Fensters zusätzlich zum runden Fenster bei der Reservetherapie der akuten idiopathischen Ertaubung einen Vorteil?

BACKGROUND: Following sudden unilateral deafness or severe sensorineural hearing loss, patients with unsuccessful intravenous steroid therapy can be treated with explorative tympanotomy with sealing of the round (RW) and/or oval window (OW), due to suspected rupture of the RW with perilymph fistula (PLF) or a fissula ante fenestram (FAF). This study investigated whether additional sealing of the oval window (RW+OW) achieved an improved hearing benefit as compared to sealing of the round window only (RW) . METHODS: This retrospective study investigated 54 patients with acute profound hearing loss who underwent tympanoscopy. Audiometric examinations were performed preoperatively and at two postoperative intervals (1 month and 3-6 months after surgery). In 28 patients, the OW was sealed in addition to the RW. RESULTS: No intraoperatively visible PLF or FAF were reported. Hearing thresholds were significantly reduced in the early postoperative follow-up period and further improvement was observed 3-6 months later. No significant differences between the RW and RW+OW subgroups were seen at either follow-up timepoint. In 65% (Kanzaki criteria) and 74% (Siegel criteria) of patients, partial or complete postoperative hearing improvement was observed. Upon comparing the groups of patients with and without hearing improvement, no statistical significance was found in terms of gender, age, secondary diagnoses, or latency period between symptom onset and surgery. CONCLUSION: Additional sealing of the OW did not lead to significantly better postoperative hearing thresholds. In general, postoperative hearing improvement corresponds to published spontaneous remission rates.

Bone morphogenetic protein 2/SMAD signalling in human ligamentocytes of degenerated and aged anterior cruciate ligaments.

OBJECTIVE: Anterior cruciate ligament (ACL) degeneration leads to knee instability and favors osteoarthritis (OA) progression. During ageing the growth factor sensitivity of ligaments changes but nothing is known about BMP2-signalling and -sensitivity in degenerated ACLs. This study addressed the question whether a dysregulated BMP2 signalling might contribute to age- and OA-dependent ACL degeneration. METHOD: ACL samples from patients with/without OA of different ages (<60 and ≥60 years, males, females) were graded histopathologically (n = 45). After stimulation of cultured ACL fibroblasts with 5 nM BMP2 for different time points, phosphorylation of SMAD1/5/8 and gene expression of crucial BMP2 signalling proteins, ligamentogenic and chondrogenic transcription factors, scleraxis (SCX) and SOX9, were analyzed. RESULTS: ACL samples displayed different grades of degeneration, often associated with synovitis and calcium deposits. Degeneration correlated significantly with synovitis. ACL fibroblasts expressed BMP type I receptors ALK3 and ALK6 and the BMP type II receptor BMPRII. Donors could be divided into "responders" and "non responders" since their BMP2 mediated SMAD1/5/8 phosphorylation level differed. Basal ID1 expression was lower in cells derived from OA compared with non-OA patients and BMP2 led to an ID1 induction in both. Irrespective of BMP2 stimulation, the donor age significantly influenced the expression profile of BMP6 and SCX but not BMP signalling. The BMP2-mediated SMAD6 expression differed between OA and healthy ACL fibroblasts. CONCLUSION: Our data indicate that the expression level of BMP2/SMAD target genes such as ID1 and SMAD6 was reduced in ACL fibroblasts derived from OA compared with non OA patients.

Comparison of seven commercial enzyme-linked immunosorbent assays for the detection of anti-diphtheria toxin antibodies.

Determination of immune status of patients to diphtheria toxin is based mainly on the results of commercially available ELISA kits. The aim of the present study was to compare the results obtained by ELISAs from seven different manufacturers: Mikrogen, Immunolab, Sekisui Virotech, NovaTec, Virion\Serion, IBL International and Euroimmun. All assays were performed according to the manufacturers' instructions. The concentrations of the anti-diphtheria toxin antibodies in 72 serum samples were calculated on the basis of curves constructed from standards supplied by manufacturers and the new reference material-International Standard for Diphtheria Antitoxin (10/262). The repeatability and reproducibility of all the ELISA kits tested were good. Number of sera with concentrations of the anti-diphtheria toxin antibodies below the WHO-recommended level of protection (0.1 IU/ml) were dependent on the ELISA used: Mikrogen, 20/72 samples (27.7%); Immunolab, 11/72 samples (15.3%); Sekisui Virotech, 0/72 samples (0%); NovaTec 18/72 samples (25.0%); Serion 12/72 samples (16.7%); IBL International, 7/72 samples (9.7 %); and Euroimmun, 17/72 samples (23.6%). However, the results obtained in particular ELISAs, with the exception of Sekisui Virotech, were much more consistent when the concentrations of the anti-diphtheria toxin antibodies in 72 sera measured by using curves constructed from the International Standard 10/262. The data obtained clearly demonstrated that manufacturer-dependent differences between anti-diphtheria IgG ELISA kits exist. The differences in recommendations accepted by the individual manufacturers together with differences shown in our studies in sensitivity greatly affect the clinical interpretation of results.

[Use of recombinant P1 protein of Mycoplasma pneumoniae for the serodiagnosis of mycoplasmosis]. / Zastosowanie metody inzynierii genetycznej do uzyskania bialka P1 Mycoplasma pneumoniae oraz ocena jego przydatnosci w serodiagnostyce mykoplazmozy u ludzi.

INTRODUCTION: Mycoplasma pneumoniae is a common etiologic agent of community-acquired respiratory infection. In serological diagnosis of M. pneumoniae infections tests have been described based on the purified P1 protein, which is the most important virulence factor of this pathogen, as antigen. The aim of his study was to express and purify a recombinant protein P1 M. pneumoniae and next evaluate this protein as high specific antigen in serodiagnosis of mycoplasmosis. METHODS: Protein P1 M pneumoniae was expressing in E. coli BL21 (DE3) using the pET-30 Ek/LIC expression vector. Based on published literature, we decided to express C-terminal region [P1-C1] encompassing amino acid residues 1160 to 1521. Purification was accomplished by immobilized metal (Ni2+) affinity column chromatography (His-trap). Serum samples collected from 221 patients with mycoplasmosis, positive in complement fixation test (CFT), 87 patients with other then mycoplasmosis bacterial infections and 80 blood donors were screened for anti-P1 recombinant protein IgA, IgG and IgM antibodies by using the home-made ELISA. RESULTS: SDS-PAGE and Coomassie brilliant blue staining confirmed a high purity of the recombinant P1 protein preparation with an expected molecular mass of 39,7 kDa. The specificity of the recombinant protein was confirmed by western blot analysis using serum samples from rabbits immunized by M pneumoniae. The results of ELISA revealed that more then 70.0% of patients with mycoplasmosis confirmed by CFT, had antibodies to recombinant P1 protein in diagnostically significant level (x + 2SD). The antibodies were found only sporadically in sera obtained from patients with other then mycoplasmosis bacterial infections and clinically healthy persons. A comparison of results obtained in home-made ELISA with results of commercial western blot (Virotech) showed similar, ranged from 84.2% to 97.4%, compatible of results. The IgM antibodies to recombinant P1 protein were found in 87.2% sera obtained in acute phase of disease, in 80.0% sera obtained 2-4 weeks after onset of clinical symptoms and only in 43.8% sera obtained in chronic mycoplasmosis. CONCLUSIONS: The present study confirmed the earlier observations of the high usefulness of recombinant P1 protein for reliable serologic diagnosis of M. pneumoniae infection.

Susceptibility of Polish clinical strains of Yersinia enterocolitica serotype O3 to antibiotics.

A total of 199 clinical strains of Yersinia enterocolitica serotype O3, biotype 4 were tested for their susceptibility to antibiotics (158 strains carried the virulence plasmid pYV and 41 strains did not). A total of 114 isolates were tested by a standard disk diffusion method for 21 antibiotics. Almost all strains tested were resistant to ampicillin and cefazolin and susceptible to amoxycillin/clavulanate, cefaclor, cefamandole, cefuroxime, cefotaxime, ceftriaxone, aztreonam, imipenem, gentamicin, amikacin, netilmicin, tetracycline, doxycycline, chloramphenicol, ciprofloxacin, sulphamethoxazole, trimethoprim, co-trimoxazole and furazolidone. In addition, minimal inhibitory concentrations of 15 antibiotics were determined by the agar dilution method for all 199 strains (158 carrying plasmid pYV and 41 strains that did not). Third-generation cephalosporins such as cefotaxime and ceftriaxone and a fluoroquinolone (ciprofloxacin) were the most active antimicrobial agents tested followed by aztreonam, imipenem, trimethoprim, tetracycline, gentamicin, chloramphenicol, amoxycillin/clavulanate, cefaclor, cefuroxime, amikacin, furazolidone and sulphamethoxazole. The present study demonstrated a high susceptibility of clinical strains of Y. enterocolitica to most of the tested antibiotics. In general there was no significant difference between susceptibility to antibacterial agents of strains with or without plasmid pYV.

Epidemiology of Mycoplasma pneumoniae infections in Poland : 28 years of surveillance in Warsaw 1970-1997.

Mycoplasma pneumoniae is a common cause of lower respiratory tract disease in humans, particularly among older children, adolescents, and young adults. Infections are endemic in cities and epidemic increases are observed at intervals of 4 to 7 years. M. p

[Electrophoretic and immunologic comparative analysis of Mycoplasma pneumoniae and Mycoplasma genitalium proteins]. / Elektroforetyczna i immunologiczna analiza porównawcza komórkowych bialek Mycoplasma pneumoniae i Mycoplasma genitalium.

The Mycoplasma pneumoniae FH strain routinely used in our laboratory for over 25 years as antigen in serological tests, 2 reference M. pneumoniae strains from ATCC (29342 and M129) and 3 isolates of M. pneumoniae obtained in 1995 from pneumonia patients were compared by SDS-PAGE, complement fixation test (CFT) and by Western-immunoblotting against human and rabbit serum samples with high level of mycoplasmal antibodies. On SDS-PAGE all M. pneumoniae strains showed the same number of 23 polypeptides on the gel with identical molecular weights. The same strains on immunoblotting against human and rabbit serum samples showed six bands: 170, 89, 75, 55, 38 and 33 kDa with the strongest antibody staining in 170-(P1 protein) and 89-kDa bands. Because of its known antigenic relationships Mycoplasma genitalium was used for comparison. The pattern of M. genitalium proteins on SDS-PAGE was similar to pattern of M. pneumoniae but distinguishable. On immunoblotting six proteins of M. genitalium (135, 127, 110, 95, 75 and 45 kDa) reacted with human and rabbits immunoglobulins for M. pneumoniae antigens. Furthermore in complement fixation test both antigens, prepared from M. pneumoniae and M. genitalium, reacted as well with human and rabbit immunoglobulins for M. pneumoniae and with rabbit immunoglobulins for M. genitalium. These cross-reactions observed in serological techniques could give false positive results in routine diagnosis of M. pneumoniae infections. In such situations showing on immunoblott of presence in tested serum sample of antibodies to 170- and 89 kDa proteins could confirm M. pneumoniae infection.

[Latex test used for diagnosis of dysentery caused by Shigella sonnei]. / Test lateksowy w zastosowaniu do diagnostyki czerwonki wywolanej przez S. sonnei.

The aim of this study was to evaluate the usefulness of latex reagent coated with immunoglobulins specific for antigens of phase I and II of S. sonnei for detection of these antigens in primary, mixed bacterial cultures. The study was performed on 919 fecal samples from individuals with clinical symptoms of dysentery, convalescents and from contact individuals. Material used for the test was bacterial suspension collected from McConkey or SS agars and a culture from selenite F broth heated at 100C. The results of the latex test were compared with the results of isolation of S. sonnei from the same cultures. S. sonnei was isolated from feces of 140 individuals (15.2%), while the latex test was positive in 215 cases (23.4%). The highest testing effectiveness , significantly higher than when isolation of pathogen was performed, was obtained only when 18-20 hr culture on Selenite F medium was used for latex test. The correlation between efficacy of testing for S. sonnei and phosphate content of Selenite F and a mode of its preparation was found. The latex test allows to eliminate from further bacteriological studies cultures free of S. sonnei thus it gives measurable economical profits and it shortens significantly time period of bacteriological examination.

[Use of heterophilic mononucleosis antigen in the latex diagnostic test. III. Diagnostic field studies]. / Heterofilny antygen mononukleozowy w zastosowaniu do lateksowego testu diagnostycznego. III. Terenowe badania diagnostyczne.

The aim of this study was to evaluate usefulness of latex coated with a preparation of heterophilic mononucleosis antigen for the detection of antibodies present during a course of infectious mononucleosis. Studies were performed in district serological laboratories. Sanitary-Epidemiological Stations in conditions of routine diagnostics. Studies were performed on 656 blood serum samples collected from individuals with a clinical suspicion of infectious mononucleosis. Latex and Paul-Bunnell-Davidsohn (PBD) tests were run in parallel. Out of 154 blood serum samples which contained antibodies detected by PBD test in diagnostically significant titer of 1:56 or higher, 151 were reactive in latex test in the dilution 1:5 or higher, while in the remaining three cases agglutination appeared with undiluted serum only. Out of 268 serum samples tested in latex test 167 reacted in 1:5 titer - diagnostically accepted as a significant or in higher titers. The sensitivity of latex test amounted to 98.1% and specificity was 96.9% as compared to reference Paul-Bunnell-Davidsohn test. The results of our study suggest the possibility of replacing Paul-Bunnell-Davidsohn test by latex test performed as a method providing the possibility of determination of the presence of heterohilic antibodies in blood serum samples and the follow up of their dynamics.

[Heterophile infectious mononucleosis antigen used in the latex diagnostic test. I. A method of antigen isolation and its serologic activity]. / Heterofilny antygen mononukleozowy w zastosowaniu do lateksowego testu diagnostycznego. I. Metodyka uzyskiwania antygenu i jego serologiczna aktywnosc.

The aim of this study was to elaborate a method of heterophile mononucleosis antigen preparation useful for latex coating. This antigen was isolated from bovine red blood cells stroma by the technique of Schwarzweiss and Tomcsik with author's own modification, in which introductory extraction of erythrocytes stroma ++ was performed by means of trichloracetic acid, aqueous extraction and elution of active substance with 80% ethanol. Besides of heterophile antigen preparation obtained by the method of Schwerzweiss and Tomcsik (preparation S-T) two serologically++ active preparations were obtained (fraction I and IV), which ability to inhibit PBD agglutinating reaction and bovine red blood cells haemolysis was 16 and 8 times lower, respectively, than S-T preparation. The preparation of heterophile mononucleosis antigen obtained differed in latex coating efficacy. In order to prepare latex reagent MZ-I (from fraction I) a solution of preparation of 125 micrograms/ml concentration was used, for MZ-II (from fraction IV)--50 micrograms and for MZ-III (from preparation S-T)--15 micrograms/ml. The reagent MZ-I showed, the highest activity in agglutinating test with human serum containing heterophile mononucleosis antibodies while two others reacted with 2-4 times lover serum dilutions. Similar differentiated reactivity with these reagents was found in latex test with 15 sera from patients suspected of having infectious mononucleosis.

[Heterophilic infectious mononucleosis antigen used in the latex diagnostic test. II. Preliminary evaluation of the usefulness of latex reagents for detection of serum heterophile antibodies in patients with infectious mononucleosis]. / Heterofilny antygen mononukleozowy w zastosowaniu do lateksowego testu diagnostycznego. II. WSTepna ocena przydatnosci odczynników lateksowych do wykrywania surowiczych przeciwcial heterofilnych u chorych na mononukleoze zakazna.

The aim of this study was to evaluate the usefulness of laboratory made reagent of polystyrene latex coated with three preparations of mononucleosis antigen (reagent MZ-I, MZ-II, and MZ-III) for detection of heterophile antibodies in patients sera with clinical symptoms of infectious mononucleosis. These studies were carried out on 153 serum samples taken from persons suspected of being infected with EB virus and on 100 healthy controls--blood donors. The results of latex tests were compared to the results of Paul-Bunnell-Davidsohn (PBD) and hemolytic tests. Out of three latex reagents evaluated only one (reagent MZ-I) showed sensitivity equal to PBD and haemolytic tests. The sensitivity reached 91.1%. Agglutination reaction when appeared in latex test at serum dilution 1:5, is considered positive and diagnostically significant result.

[Evaluation of the usefulness of commercial enzyme-linked immunosorbent assays for diagnosis of Mycoplasma pneumoniae infections]. / Ocena przydatnosci komercyjnych testów immunoenzymatycznych i testu opracowanego we wlasnym zakresie do diagnostyki zakazen wywolywanych przez Mycoplasma pneumoniae.

The usefulness of the ELISA distributed by BioChem ImmunoSystems, Medial Polska, Biomedica/Virotech and prepared in our laboratory (ELISA FH-K) for diagnosis of the M. pneumoniae infections was estimated. Eighty six serum samples obtained from 86 patients with respiratory tract infections were simultaneously tested by ELISA-IgM/-IgG and by complement fixation test which was accepted as a reference test. The highest sensitivity in relation to the CFT was displayed by the ELISA BioChem ImmunoSystems and Medial Polska (100%), slightly lower sensitivity by the ELISA Biomedica/Virotech--96.5% and ELISA FH-K--90.9% when determining mycoplasmal antibodies of IgM. The lowest sensitivity was displayed by the ELISA Biomedica/Virotech when determining antibodies of the IgG class (54.9%). The specificity of ELISA in relation to the CFT was generally higher when detecting mycoplasmal antibodies of the IgM class then of IgG class. The study demonstrated that all 4 ELISA may be used in routine serodiagnosis of M. pneumoniae infection. For the improve of sensitivity of ELISA it's recommended to measure simultaneously the level of mycoplasmal antibodies of IgM and IgG.

[Comparative evaluation of latex reagents of Wellcolex Color Salmonella and Latex Salmonella used for detection and identification of Salmonella rods]. / Porównawcza ocena zestawów lateksowych Wellcolex Colour Salmonella i Lateks Salmonella sluzacych do wykrywania i identyfikacji somatycznych antygenów paleczek z rodzaju Salmonella.

The aim of this study is to compare the sensitivity and specificity of the Wellcolex Colour Salmonella (WCS) set produced by Murex with the Latex Salmonella (LS) set developed by the Polish company Biomex and commonly used in sanitary and epidemiological stations. An attempt is also made to determine the possible usefulness of the WCS test in routine diagnostic of salmonellosis and of S. typhi carrier. The sensitivity and specificity of latex reagents of both sets were determined basing on reactions with the suspensions of 17 selected Salmonella strains representing the A-E and G serological groups and with Salmonella O antigen preparations obtained using the Boivin method. Both reagents from the WCS set were found to detect Salmonella bacteria in a suspension having a minimal density of 7.5-60 x 10(7) cfu/ml while both the polyvalent reagent B-E and monovalent reagents from the LS set still reacted with suspensions of 0.47-15 x 10(7) cfu/ml density. Comparison of the sensitivity of both tests basing on reactions with Salmonella O antigen specimens from the B to E groups revealed that the latex reagents from the two sets detected antigens from the C2 and C3 groups and the E group in concentrations of 1 microgram/ml and 0.5 microgram/ml respectively. In the case of antigen specimens from group B, group C and group D, the LS test detecting these antigens in concentrations of 0.25-1 microgram/ml turned out to be four to eight times more sensitive in reaction with a polyvalent agent and two to eight times more sensitive in reaction with monovalent reagents than the WCS set. The evaluated latex reagents from the WCS set and LS set reacted specifically with both cell suspensions and Salmonella antigen preparations. Also, the applicability of the two latex sets to detect and identify Salmonella antigens in liquid and mixed bacterial cultures of these germs in selenite broth was compared. A positive result of the WCS test was obtained in 41% of Salmonella culture samples whereas Salmonella antigens were found in all the studied culture samples when the polyvalent reagent B-E from the LS set was used and in 97.5% of the samples when monovalent reagents were used. The study showed that in spite of the comparable specificity of the WCS test with respect to the LS set produced in Poland, the latex reagents from the WCS set turned out to be of little use in detecting and identifying Salmonella antigens in mixed bacterial cultures in selenite broth.

[A trial application of the latex test for evaluation of antibody level in rabbit immune sera]. / Próba wykorzystania odczynu lateksowego do oceny poziomu przeciwcial w króliczych surowicach odpornosciowych.

A latex test was elaborated which served for evaluation of quality of rabbit immune sera for antigen 0 of selected Gram-negative bacteria. Sensitivity and specificity of this test in comparison with passive hemagglutination and immunoenzymatic DOT-ELISA reactions was evaluated. These studies were performed on immune sera for antigen O of Salmonella groups B, C1, C2, D and E, Yersinia pseudotuberculosis and in antigen preparations for above listed microorganisms both in homologous and heterologous systems. It was found that sensitivity of the latex test is 9 to 160 times lower than that of passive hemagglutination and 7 to 307 lower than for DOT-ELISA. Sensitivity of the latex test and passive hemagglutination reaction was evaluated on the basis of results of cross reaction between studied antigens and unabsorbed rabbit sera, establishing so called sensitivity indexes, which were informing how many times heterologous titer is lower than homologous titer. So evaluated sensitivity of the latex test was close to sensitivity of the passive hemagglutination reaction. It was found that slide latex test is characterized by satisfactory sensitivity and good sensitiveness and may be used for evaluation of antibody level 0 antigens of Salmonella and Yersinia. The value of this test is characterized by high repeatability of results, as well as low work and time-consuming.

[Evaluation of usefulness of OWD and OIEP in diagnosis of epidemically occurring infections caused by Mycoplasma pneumoniae]. / Ocena przydatnosci OWD i OIEP w diagnostyce epidemicznie wystepujacych zakazen wywolanych przez Mycoplasma pneumoniae.

Blood serum samples of 3593 persons clinically suspected of infection with Mycoplasma pneumoniae were tested. Of these, patients with pneumoniae constituted 66.5%, upper respiratory tract infection--24.0% and with symptoms localized outside the respiratory system--9.5%. These studies were performed by application of complement fixation test (OWD) and immunoelectroprecipitation (OIEP) methods, accepting as a diagnostically significant--titer 1:60 or higher and/or occurrence in OIEP reaction with serum diluted 1:2 or more. Among patients studied prevailed children in the age of 3 to 16 years (61.6%). Mycoplasmosis was detected in these patients in 1071 out of 2236 cases (47.9%). Compatible results in both tests were obtained in 90.6% patients, whereas OWD only in 3.0% and OIEP only in 6.4% cases. Simultaneous application of both tests increased detectability of infections caused by M. pneumoniae by 3% in relation to OIEP and by 6.4% in relation to OWD.

[Evaluation of the reliability of serological diagnostic tests for Mycoplasma pneumoniae infection carried out by laboratories in sanitary epidemiological stations (WSSE)]. / Ocena wiarygodnosci wyników serologicznych badan diagnostycznych w kierunku zakazen Mycoplasma pneumoniae wykonywanych w laboratoriach WSSE.

In the action undertaken for evaluating of the reliability of serological tests for M. pneumoniae infection 33 laboratories of the Sanitary Epidemiological Stations participated. Every laboratory had to determine twice at an interval of 2-4 weeks the level of mycoplasma antibodies by the complement fixation test in serum samples divided into 4 groups: sera not containing these antibodies-titre < 5, or containing them in titres of 60, 120 and 320. The correct results of the determinations were obtained in 27 laboratories (81.8%) for samples without M. pneumoniae antibodies, and in 22 (66.6%) and 14 (42.4%) for samples with titres of 60 and 120, and 320 respectively. Only 4 laboratories (12.1%) obtained correct results of these determinations for every sample and in both testing series. In these series considered separately correct results were obtained in 9 (27.3%) and 8 (24.2%) laboratories. Faulty results in all samples in both testing series were reported from 2 (6.1%) laboratories. In the individual series all false results were obtained in 4 (12.1%) and 3 (9.1%) laboratories. The study showed that for raising of the quality and reliability level of serological investigations for M. pneumoniae infection a permanent practice should be periodic training of laboratory workers and frequently repeated interlaboratory controls of the reliability of test results.

[Use of PCR methods for detection of selected genes of Mycoplasma pneumoniae]. / Zastosowanie metody PCR do wykrywania wybranych genów Mycoplasma pneumoniae.

The aim of this study was standardization of PCR for the detection of gene encoding the P1 protein, 16S rRNA and elongation factor Tu of M. pneumoniae. A total of 13 strains of M. pneumoniae, 28 strains of other mycoplasmas and 14 strains of different bacteria causing respiratory tract infections were tested. In all of tested M. pneumoniae strains the presence of the sought genes was confirmed. The specificity of DNA was confirmed by the restriction endonuclease analysis with enzymes Hind III, Alu I and Hha I. With none of primers specific for the M. pneumoniae genes amplification of DNA from other bacteria was noted. The PCR method with the selected primers allowed to detect from 10(2) to 10(4) cfu M. pneumoniae/ml suspended in broth. The obtained results indicate that the PCR method can be used for detection of M. pneumoniae genes. A very good sensitivity and specificity predestine++ PCR as a potential quick diagnostic method for identification of M. pneumoniae in clinical specimens.

[Evaluation of the usefulness for the latex agglutination test for serodiagnosis of Mycoplasma pneumoniae infections]. / Ocena przydatnosci lateksowego testu aglutynacyjnego do serologicznej diagnostyki zakazen wywolywanych przez Mycoplasma pneumoniae.

The usefulness of latex agglutination test prepared in our laboratory for the diagnosis of M. pneumoniae infections was assessed. A total of 628 serum samples obtained from patients with respiratory tract infections were tested by complement fixation test and by latex test, from among them 274 serum samples were additionally tested by ELISA--Ig A/--IgG/--IgM and by immunoelectroprecipitation test. The highest sensitivity and specificity was displayed by the latex test in relation to ELISA when determining mycoplasmal antibodies of IgM class (respectively 82.1% and 89.6%) and to the complement fixation test (81.0% and 89.0%). Positive latex test results in our investigations were associated only with the presence of IgM antibodies and were not dependent on the IgA and IgG antibody classes. The latex agglutination test may be used in routine serodiagnosis of mycoplasmosis under condition that the results obtained in this test will be confirmed by the complement fixation test or ELISA.

[Identification of enteropathogenic strains of E. coli (EPEC) by application of the latex test. I. Evaluation of specificity and sensitivity of the test]. / Identyfikacja enteropatogennych szczepów E. coli (EPEC) przy zastosowaniu testu lateksowego. I. Ocena swoistosci i czulosci testu.

The study was aimed at elaboration of the test and evaluation of specificity and sensitiveness of it during routine diagnosis and detection of enteropathogenic strains of E. coli (EPEC), basing on direct identification of O antigen of these bacteria. The studies was performed by application of polystyrene latex having particles of 0.8 micron in diameter and coated with rabbit globulins immune for O antigens of 14 serotypes of E. coli identified in Poland as EPEC strains. Fourteen univalent reagents and three multivalent reagents (identifying 4-6 group antigens) were used. Latex reagents, prepared by own method, reacted only with suspensions of homologous strains of E. coli, not agglutinating in presence of O antigens of remaining EPEC strains. Evaluating specificity of univalent reagents against suspensions of bacteria representing 111 other than EPEC strains of E. coli, it was found that only in 5 cases agglutination did occur in presence of heterologous cell suspensions. These reactions concerned strains representing 03, 04, 010, 090 and 0106 group antigens. Comparative studies performed with latex and slide agglutination tests and OK sera on 391 freshly isolated samples of feces of children (determined in regional laboratories as EPEC) have shown that as a results of serological reidentification of these strains with commercial OK sera for 14 serotypes of EPEC, positive result of agglutination test was obtained only with 277 (70.8%) of strains. These results suggest a possibility of application of latex reagents for detection of EPEC strains basing on identification of O antigen of these bacteria.

[Occurrence of p-fimbriae in enteropathogenic E coli (EPEC) and in E. coli strains isolated from patients with urinary tract infections]. / Wystepowanie fimbrii P u enteropatogennych paleczek E. coli (EPEC) oraz u szczepów E. coli wyosobnionych od osob z zakazeniem ukladu moczowego.

The aim of this study was to gain knowledge of prevalence of P+ clones among EPEC strains isolated from children with diarrhoea and E. coli strains isolated from urine. Three hundred eighty four E. coli strains isolated from children with diarrhoea were tested. They belonged to 11 serotypes (018, 025, 026, 044, 055, 0111, 0114, 0119, 0124, 0125, and 0128). Nine hundred thirty colonies of E. coli from Mac Conkey's agar plated quantitatively with urine samples of 178 individuals suffering from urinary tract infections were also tested. All strains were assayed by mannose-resistant active haemagglutination test (MRHA) and by slide agglutination using self prepared latex reagent for detection of P fimbriae. Out of 384 E. coli strains tested 122 (31.8%) showed presence of adhesins detected by mannose-resistant active haemagglutination test (MRHA) and in 90 (23.3%) out of all tested strains the presence of P fimbriae was found. The highest percentage of P fimbriae prevalence was found in E. coli belonging to the following serotypes: 018 (in 68.9% strains), 025 (in 29.2% strains), and 0125 (in 25.0% strains). This type of fimbriae was also detected in serotypes 026 (9.1%), 044 (8.7%), 055 (5.6%), and 0119 (in 2 strains out of 5 isolated). Out of 933 colonies of E. coli, isolated from 178 urine samples, 434 (46.5%) colonies gave positive results in MRHA test, including 133 positive in latex test for P fimbriae. These studies showed that for MRHA adhesins, including P fimbriae, a parallel examination of higher number of E. coli was necessary.(ABSTRACT TRUNCATED AT 250 WORDS)