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P1849 Molecular typing of clinical isolates of Escherichia coli by a new PCR MP method
Molecular typing in clinical bacteriology in released phage DNAs. These results suggest that the population of Fvir-lacking functional prophages is vast and that the polylysogeny would be the product of the accumulation of temperate phages that eventually become defective. P1848 Molecular epidemiology of Streptococcus pneumoniae associated with paediatric invasive disease in the Czech Republic H. Zemlickova, V. Jakubu, P. Urbaskova, M. Musilek, J. Motlova (Prague, CZ) Objectives: To characterise invasive isolates of S. pneumoniae associated with paediatric invasive disease in Czech children under 6 years of age by multilocus sequence typing (MLST) and to study the clonality of the causative serotypes collected prior to the introduction of the pneumococcal conjugate vaccine into the Czech Republic. Methods: The strains (n = 124) isolated from blood or cerebrospinal fluid of children under 6 years of age between 1996 and 2003 were referred to the National Institute of Public Health by 39 microbiology labs from 28 cities. Serotyping was performed by Quellung reaction. The minimal inhibitory concentrations (MIC) of penicillin, erythromycin, tetracycline and chloramphenicol were determined by the CLSI broth microdilution method. MLST typing was carried out for all available isolates (n = 80) of the major predominant serotypes 6B, 14, and 23F. Results: Overall, 5 (4.0%) strains exhibited reduced susceptibility to penicillin but only one of these was highly resistant. The resistance rates to erythromycin, tetracycline and chloramphenicol were 5.6%, 8.1% and 4.8%, respectively. The most frequent serotypes were 6B (19), 14 (15), 23F (12), 19F (11), 9V (9), 18C (9) and 1 (8). MLST typing was performed on 80 (64.5%) viable isolates of the above named serotypes. Nine of 32 revealed sequence types (STs) were newly identified. Isolates of serotype 6B were quite diverse: 17 strains yielded 8 clonal lineages (10 STs). In contrast, isolates of serotypes 1, 9V and 14 were highly homogenous. Serotypes 1 and 9V were classified into a single clone each. Thirteen of 15 isolates of serotype 14 had the same sequence type (ST124). Conclusion: The vaccine-included serotypes 6B, 14, 23F, 19F, 9V and 18C are most frequently associated with paediatric invasive disease in the Czech Republic. Clonality varies between serotypes. The continued presence of previously described invasive clones of S. pneumoniae was demonstrated for serotypes 1, 9V and 14. P1849 Molecular typing of clinical isolates of Escherichia coli by a new PCR MP method B. Krawczyk, A. Sledzinska, A. Samet, M. Bronk, J. Komarnicka, J. Leibner, J. Kur (Gdansk, PL) Background: Epidemiological typing of bacteria is a routine procedure used in the investigation of infectious outbreaks and requires accurate molecular typing methods with high discriminatory power. We had performed evaluation of a new PCR melting profile (PCR MP) technique for E. coli strain differentiation. Results were compared with PFGE method. Methods: Epidemiologic investigation was shown for E. coli isolates from patients of the Hematological Unit of Clinical Hospital in Gdansk. A total of 90 isolates from 36 patients were included. The E. coli were recovered from blood (66), stool (15), urine (8) of patients with suspected bloodstream infection. To study the genetic similarities between E. coli isolates, a new PCR MP method and REA-PFGE were used. Results: PCR MP analysis distinguished 44 types (H1-H44) among all isolates studied. One genotype was markedly predominant (H2), as this was represented by 10 isolates from 4 patients. This genotype probably represents E. coli strain from hospital infection. The molecular typing by REA-PFGE found also 44 unique profiles. Clustering of REAPFGE fingerprinting data matched exactly PCR MP data. The PCR MP was next investigated for the similarities of the E. coli isolated from different sites of examined patients. PCR MP fingerprinting analysis exhibited usefulness for bacterial spread monitoring. As expected, the
S529 related epidemiologically isolates show a high degree of similarity. Genotype similarity was documented in 10 of 36 patients with E. coli bacteraemia for whom paired blood and faecal or urine isolates were available for genomic typing, and probably was the major source of these bacteraemias. Conclusion: We found that PCR MP technique is a rapid method that offers good discriminatory power, excellent reproducibility and may be applied for epidemiological studies. We suggested that there is at least a similar power of discrimination between the present gold-standard REAPFGE and a PCR MP method. Data presented here demonstrate the complexity of the epidemiological situation concerning E. coli that may occur in a hospital.
P1850 The analysis Y. pestis strains from Kazakhstan and USA by PFGE method T. Meka-Mechenko, R. Tashmetov, B. Atshabar, L. Nekrassova (Almaty, KZ) Objectives: Y. pestis strains from plague foci of Republic of Kazakhstan differ on phenotypic properties. The differences in plasmid profile and structure of genes are marked also. The comparative analysis Y. pestis strains isolated from the natural plague foci of Kazakhstan and USA by PFGE method are of interest. Methods: Are investigated by PFGE method Y. pestis 48 strains isolated in 1938–2002 from the natural plague foci of Kazakhstan and 12 strains from USA. Strains are isolated from fleas, ticks, wild rodents, domestic animals and from the plague patients. Results: Y. pestis strains isolated from autonomous plague of Central Asian desert natural focus, Volga-Ural steppe autonomous focus, VolgoUral sandy natural focus have identity at the genetic level from 82.6% up to 100%, biotype of these strains is mediaevalis. Only 1 strain from Ustyurt autonomous focus (strain’s biotype is antigua) is generically close (84.5%) to strain from Tian-Shanian natural plague focus (Sarydzhas autonomous focus). Two plague strains by genetic structure are closer to strains isolated in USA (83%) in comparison with others strains isolated from plague foci of Kazakhstan (77.2%). In strains isolated from Sarydzhas autonomous focus (Tian-Shanian natural focus) percent of similarity rather low – 77.2%. One strain with positive fermentation of glycerine and negative denitrification 95.7% of similarity of genetic picture with strain isolated from Moinkum autonomous focus (Central Asian desert natural plague focus). Others investigated strains are typical and percent of genetic identity makes 84.5%. The similarity of genetic structure strains isolated from USA varies from 88.8% up to 100%. Rather low percent of identity plague strains isolated in plague foci of Kazakhstan (77.2% up to 100%), is caused landscape-epizootological feature of sites and characteristic properties of circulating populations of the activator of plague. Conclusion: The identity plague strains on genetic level depend on geographical region and from strain’s biotype. In strains concerning to one biotype and isolated from one natural plague focus percent of identity at molecular level is high. The received results of computer analysis and dendrogram of phylogenetic connections between investigated strains allow to determine their origin that increases efficiency of epidemiological of supervision of plague.
P1851 Molecular diversity of Bacillus anthracis in Kazakhstan A. Aikimbayev, L. Lukhnova, S. Zakaryan, G. Temiraliyeva, Y. Pazylov, T. Meka-Mechenko, W. Easterday, M. Van Ert, P. Keim, T. Hadfield, S. Francesconi, J. Blackburn, M. Hugh-Jones (Almaty, KZ; Flagstaff, Palm Bay, Washington D.C., Baton Rouge, US) Objectives: In the central Asian republic of Kazakhstan, anthrax is endemic and represents a public health concern. In this study we used high resolution genotyping to examine Bacillus anthracis strain dynamics among historical outbreaks in Kazakhstan and to understand Kazakh B. anthracis genetic diversity on a regional and global scale.
S529 related epidemiologically isolates show a high degree of similarity. Genotype similarity was documented in 10 of 36 patients with E. coli bacteraemia for whom paired blood and faecal or urine isolates were available for genomic typing, and probably was the major source of these bacteraemias. Conclusion: We found that PCR MP technique is a rapid method that offers good discriminatory power, excellent reproducibility and may be applied for epidemiological studies. We suggested that there is at least a similar power of discrimination between the present gold-standard REAPFGE and a PCR MP method. Data presented here demonstrate the complexity of the epidemiological situation concerning E. coli that may occur in a hospital.
P1850 The analysis Y. pestis strains from Kazakhstan and USA by PFGE method T. Meka-Mechenko, R. Tashmetov, B. Atshabar, L. Nekrassova (Almaty, KZ) Objectives: Y. pestis strains from plague foci of Republic of Kazakhstan differ on phenotypic properties. The differences in plasmid profile and structure of genes are marked also. The comparative analysis Y. pestis strains isolated from the natural plague foci of Kazakhstan and USA by PFGE method are of interest. Methods: Are investigated by PFGE method Y. pestis 48 strains isolated in 1938–2002 from the natural plague foci of Kazakhstan and 12 strains from USA. Strains are isolated from fleas, ticks, wild rodents, domestic animals and from the plague patients. Results: Y. pestis strains isolated from autonomous plague of Central Asian desert natural focus, Volga-Ural steppe autonomous focus, VolgoUral sandy natural focus have identity at the genetic level from 82.6% up to 100%, biotype of these strains is mediaevalis. Only 1 strain from Ustyurt autonomous focus (strain’s biotype is antigua) is generically close (84.5%) to strain from Tian-Shanian natural plague focus (Sarydzhas autonomous focus). Two plague strains by genetic structure are closer to strains isolated in USA (83%) in comparison with others strains isolated from plague foci of Kazakhstan (77.2%). In strains isolated from Sarydzhas autonomous focus (Tian-Shanian natural focus) percent of similarity rather low – 77.2%. One strain with positive fermentation of glycerine and negative denitrification 95.7% of similarity of genetic picture with strain isolated from Moinkum autonomous focus (Central Asian desert natural plague focus). Others investigated strains are typical and percent of genetic identity makes 84.5%. The similarity of genetic structure strains isolated from USA varies from 88.8% up to 100%. Rather low percent of identity plague strains isolated in plague foci of Kazakhstan (77.2% up to 100%), is caused landscape-epizootological feature of sites and characteristic properties of circulating populations of the activator of plague. Conclusion: The identity plague strains on genetic level depend on geographical region and from strain’s biotype. In strains concerning to one biotype and isolated from one natural plague focus percent of identity at molecular level is high. The received results of computer analysis and dendrogram of phylogenetic connections between investigated strains allow to determine their origin that increases efficiency of epidemiological of supervision of plague.
P1851 Molecular diversity of Bacillus anthracis in Kazakhstan A. Aikimbayev, L. Lukhnova, S. Zakaryan, G. Temiraliyeva, Y. Pazylov, T. Meka-Mechenko, W. Easterday, M. Van Ert, P. Keim, T. Hadfield, S. Francesconi, J. Blackburn, M. Hugh-Jones (Almaty, KZ; Flagstaff, Palm Bay, Washington D.C., Baton Rouge, US) Objectives: In the central Asian republic of Kazakhstan, anthrax is endemic and represents a public health concern. In this study we used high resolution genotyping to examine Bacillus anthracis strain dynamics among historical outbreaks in Kazakhstan and to understand Kazakh B. anthracis genetic diversity on a regional and global scale.