Sequence characterization of Haemophilus influenzae isolates in the Czech Republic in 2001–2009
Authors:
V. Lebedová 1; M. Musílek 2; P. Křížová 2
Authors‘ workplace:
Národní referenční laboratoř pro hemofilové nákazy, Oddělení vzdušných bakteriálních nákaz, Státní zdravotní ústav
1; Oddělení vzdušných bakteriálních nákaz, Státní zdravotní ústav
2
Published in:
Epidemiol. Mikrobiol. Imunol. 59, 2010, č. 4, s. 189-196
Overview
Objective:
Multilocus sequence typing (MLST)-based clonal analysis of a large set of Haemophilus influenzae strains isolated from both invasive and non-invasive cases in the Czech Republic in 2001–2009.
Material and methods:
A total of 207 H. influenzae strains, i. e. 153 H. influenzae b (Hib) strains, 2 H. influenzae e strains, 2 H. influenzae f strains and 50 non-capsulated H. influenzae (HiNT) strains, were tested by the MLST method.
Results:
MLST classified 73% of 153 Hib strains to sequence type (ST) ST-6 and 15% of them to related ST-83. Other sequence types were uncommon (ST-92, ST-95, ST-108, ST-190 and ST-326). Five new STs were found (ST-663 to ST-667). All STs observed in Hib strains belonged to clonal complex cc6. Of 12 strains isolated from clinical specimens of actual Hib vaccine failure cases, nine were classified by MLST into ST-6, two into ST-83 and one into ST-190. The H. influenzae type e isolates were of ST-18 while H. influenzae type f isolates belonged to ST-124. A HiNT strain isolated from the cerebrospinal fluid was of ST-6, typical of Hib, and may be a Hib strain that lost the capsule-forming ability. Characterization of other 49 HiNT strains revealed 37 different STs, with ST-276 being the most common (4 strains), followed by ST-556 (3 strains); other STs were only found sporadically. Nine new STs were identified (ST-668 to ST-676). STs of H. influenzae non-b are heterogeneous and do not belong to complex cc6.
Conclusion:
The population of the Czech Hib isolates is highly homogeneous and all 12 STs detected are members of the same clonal complex, cc6. ST-6 is the central genotype in Hib strains of MLST complex cc6. The detected ST-6 single-locus variants are from clonal complex cc6 (ST-83, ST-92, ST-95, ST-108, ST-190, ST-326, ST-663 to ST-667). Clonal complex cc6 was found in all vaccine failure cases. The population of H. influenzae non-b is highly heterogeneous and none of the 39 STs found matches clonal complex cc6.
Key words:
H. influenzae – clonal analysis – multilocus sequence typing – MLST – molecular epidemiology.
Sources
1. Křížová, P., Kalmusová, J., Lebedová, V., Felsberg, J., Haugvicová, R. Charakterizace kmenů Haemophilus influenzae metodou multilokusové sekvenační typizace. Epidemiol. Mikrobiol. Imunol., 2004, 53, 2, p. 74–77.
2. Lebedová, V., Beneš, Č., Kalmusová, J., Křížová, P. Závažná onemocnění způsobená Haemophilus influenzae v České republice v roce 2009. Zprávy EM, 2010, 19, 1–2, p. 31–34.
3. Maiden, M. C., Bygraves, J. A., Feil, E., Morelli, G. et al. Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. Proc. Natl. Acad. Sci. USA, 1998, 95, 6, p. 3140–3145.
4. Meats, E., Feil, E. J., Stringer, S., Cody, A. J. et al. Characterization of encapsulated and noncapsulated Haemophilus influenzae and determination of phylogenetic relationships by multilocus sequence typing. J. Clin. Microbiol., 2003, 41, 4, p. 1623–1636.
5. Metodická opatření – Metodický návod k zajištění surveillance programu invazivních onemocnění způsobených H. influenzae b. Věstník MZ ČR, prosinec 2002, částka 13, p. 15–19.
6. Metodická opatření – Zajištění surveillance programu invazivních onemocnění způsobených H. influenzae b. Věstník MZ ČR. Duben 1999, částka 3, p. 1–4.
7. Mironov, K. O., Platonov, A. E., Nikolaev, M. K., Koroleva, I. S., Shipulin, G. A. Genetic characteristic of Haemophilus influenzae type B strains isolated in Russian regions. Mikrobiol. Epidemiol. Immunobiol., 2010, 1, p. 24–28.
8. Mukundan, D., Ecevit, Z., Patel, M., Marrs, C. F., Gilsdorf, J. R. Pharyngeal colonization dynamics of Haemophilus influenzae and Haemophilus haemolyticus in healthy adult carriers. J. Clin. Microbiol., 2007, 45, 10, p. 3207–3217.
9. Oh, S. Y., Griffiths, D., John, T., Lee, Y. C. et al. School-aged children: a reservoir for continued circulation of Haemophilus influenzae type b in the United Kingdom. J. Infect. Dis., 2008, 197, 9, p. 1275–1281.
10. Platonov, A. E., Mironov, K. O., Iatsyshina, S. B., Koroleva, I. S. et al. Multilocus sequence-typing for characterization of Moscow strains of Haemophilus influenzae type b. Mol. Gen. Mikrobiol. Virusol., 2003, 2, p. 21–25.
11. Sacchi, C. T., Alber, D., Dull, P., Mothershed, E. A. et al. High level of sequence diversity in the 16S rRNA genes of Haemophilus influenzae isolates is useful for molecular subtyping. J. Clin. Microbiol., 2005, 43, 8, p. 3734–3742.
12. Sá-Lečo, R., Nunes, S., Brito-Avô, A., Alves, C. R. et al. High rates of transmission of and colonization by Streptococcus pneumoniae and Haemophilus influenzae within a day care center revealed in a longitudinal study. J. Clin. Microbiol., 2008, 46, 1, p. 225–234.
13. Schouls, L. M., van der Ende, A., van de Pol, I., Schot, C. et al. Increase in genetic diversity of Haemophilus influenzae serotype b (Hib) strains after introduction of Hib vaccination in the Netherlands. J. Clin. Microbiol., 2005, 43, 6, p. 2741–2749.
14. Skoczynska, A., Kadlubowski, M., Empel, J., Hryniewicz, W. Characteristics of Haemophilus influenzae type b responsible for meningitis in Poland from 1997 to 2004. J. Clin. Microbiol., 2005, 43, 11, p. 5665–5669.
15. Slack, M. P. E. Haemophilus. In Parker, M. T., Collier, L. H. (eds). Topley & Wilsonęs principles of bacteriology, virology and immunity. 8th ed. Philadelphia and Hamilton: B. C. Decker Inc., 1990, 2, p. 355–382, ISBN 1‑55664-290-3.
16. Tian, G., Zhang, L., Li, M., Wang, X. et al. Genotypic characteristics of Haemophilus influenzae isolates from pediatric pneumonia patients in Chengdu city, Sichuan, China. J. Microbiol., 2009, 47, 4, p. 494–497.
17. Tsang, R. S., Mubareka, S., Sill, M. L., Wylie, J. et al. Invasive Haemophilus influenzae in Manitoba, Canada, in the postvaccination era. J. Clin. Microbiol., 2006, 44, 4, p. 1530–1535.
18. Urwin, R., Maiden, M. C. Multi-locus sequence typing: a tool for global epidemiology. Trends Microbiol., 2003, 11, 10, p. 479–487.
19. Úřední věstník Evropské unie 2008/426/ES, Rozhodnutí komise ze dne 28. dubna 2008, kterým se mění rozhodnutí 2002/253/ES, kterým se stanoví definice případů pro hlášení přenosných nemocí do sítě Společenství podle rozhodnutí Evropského parlamentu a Rady č. 2119/98/ES (oznámeno pod číslem K(2008) 1589), L 159/46-159/90.
20. Vyhláška MZ ČR ze dne 6. prosince 2000, o očkování proti infekčním nemocem. Sbírka zákonů č. 439/2000, částka 121, s. 5788–5792.
21. Vyhláška MZ ČR ze dne 29. listopadu 2006, o očkování proti infekčním nemocem. Sbírka zákonů č. 537/2006, částka 174, s. 7282–7288.
22. Vyhláška MZ ČR ze dne 17. prosince 2008, o systému epidemiologické bdělosti pro vybrané infekce. Sbírka zákonů č. 473/2008, částka 151, s. 8010–8043.
23. Žemličková, H., Urbášková, P., Adámková, V., Motlová, J. et al. Characteristics of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Staphylococcus aureus isolated from the nasopharynx of healthy children attending day-care centres in the Czech Republic. Epidemiol. Infect., 2006, 134, 6, p. 1179–1187.
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Hygiene and epidemiology Medical virology Clinical microbiologyArticle was published in
Epidemiology, Microbiology, Immunology
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