A study on the genetic analysis of clinical isolates and vaccine strains of Bordetella pertussis by pulsed-field gel electrophoresis (PFGE)

Document Type: Original Articles

Authors

1 Department of Microbiology, Faculty of Veterinnary, Islamic Azad University of Karaj, Karaj, Iran

2 Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization, Tehran, Iran

Abstract

Considering the circulation of Bordetella pertussis clinical strains among populations with high vaccination coverage, it is necessary to have a proper understanding of this bacterium causing whooping cough. Various techniques, which are available for studying B. pertussis, can facilitate a proper comparison between different populations. We genotypically analyzed a collection of two vaccine strains used for the production of killed pertussis vaccine during 2000-2014 at Razi Vaccine and Serum Research Institute, Karaj, Iran. Ten clinical and two reference (Tohama 1 and 18323) strains were used by means of pulsed-field gel electrophoresis (PFGE). The genetic profiles of the vaccine master and working seeds showed no significant changes in the frequency of fingerprint types in the vaccine strains; also, the homogeneity of the profiles was demonstrated. However, the clinical isolates showed heterogeneity in the genetic profiles. In addition, serotyping was performed with monoclonal antisera to agglutinogens 2 and 3. Analysis of the fimbriae showed that all ten clinical strains expressed Fim3.

Keywords


Article Title [French]

L’analyse génétique des isolats cliniques et souches vaccinales de Bordetellapertussis par électrophorèse en champ pulsé (ECP)

Abstract [French]

Etant donnée la circulation des souches de Bordetellapertussisau sein des populations avec une couverture vaccinale élevée, une bonne compréhension de la nature de cette bactérie provoquant la coqueluche est nécessaire. Une variété de techniques ont été mises en place afin de faciliter la caractérisation des souches de Bordetellapertussisau sein des différentes populations. Une analyse génotypique a été menée sur une collection de deux souches vaccinales utilisées entre les années 2000 et 2014dans la production de vaccins inactivés de la coqueluche à l’Institut de Recherche sur les Vaccins et les Sérums Razi (Karaj, Iran). Au total, 10 isolats cliniques et 2 souches de références (Tohama 1 et 18323) ont été analysés par électrophorèse en champ pulsé (ECP). Selon nos résultats, le profil génétique de la souche-mère et des semences actives ne montre aucun changement significatif dans la fréquence des types d’empreintes observée avec les souches vaccinales. Ceci reflète donc la bonne homogénéité des profils étudiés. De plus, un sérotypage a été effectué avec des antisérums monoclonaux des agglutinogènes 2 et 3. L’analyse des fimbriae a révélé que les 10 isolats cliniques exprimaient Fim3.

Keywords [French]

  • Bordetella pertussis
  • Empreintes ADN
  • ECP
  • XbaI

Advani, A., Donnelly, D., Hallander, H., 2004. Reference system for characterization of Bordetella pertussis pulsed-field gel electrophoresis profiles. J Clin Microbiol 42, 2890-2897.

Advani, A., Hallander, H.O., Dalby, T., Krogfelt, K.A., Guiso, N., Njamkepo, E., von Konnig, C.H., Riffelmann, M., Mooi, F.R., Sandven, P., Lutynska, A., Fry, N.K., Mertsola, J., He, Q., 2013. Pulsed-field gel electrophoresis analysis of Bordetella pertussis isolates circulating in Europe from 1998 to 2009. J Clin Microbiol 422, 428-451

Caro, V., Hot, D., Guigon, G., Hubans, C., Arrive, M., Soubigou, G., Renauld-Mongenie, G., Antoine, R., Locht, C., Lemoine, Y., Guiso, N., 2006. Temporal analysis of French Bordetella pertussis isolates by comparative whole-genome hybridization. Microbes Infect 8, 2228-2235.

Cummings, C.A., Brinig, M.M., Lepp, P.W., van de Pas, S., Relman, D.A., 2004. Bordetella species are distinguished by patterns of substantial gene loss and host adaptation. J Bacteriol 186, 1484-1492.

Mooi, F.R., Hallander, H., Wirsing von König, C.H., Hoet, B., Guiso, N., 2000. Epidemiological Typing of Bordetella pertussis Isolates: Recommendations for a Standard Methodology. Eur J Clin Microbiol Infect Dis 19, 174-181.

Mooi, F.R., van Oirschot, H., Heuvelman, K., van der Heide, H.G., Gaastra, W., Willems, R.J., 1998. Polymorphism in the Bordetella pertussis virulence factors P.69/pertactin and pertussis toxin in The Netherlands: temporal trends and evidence for vaccine-driven evolution. Infect Immun 66, 670-675.

Nar Otgun, S., Durmaz, R., Karagoz, A., Esen, B., Ertek, M., 2011. Pulsed-field gel electrophoresis characterization of Bordetella pertussis clinical isolates circulating in Turkey in 2001-2009. Eur J Clin Microbiol Infect Dis 30, 1229-1236

Schouls, L.M., van der Heide, H.G., Vauterin, L., Vauterin, P., Mooi, F.R., 2004. Multiple-locus variable-number tandem repeat analysis of Dutch Bordetella pertussis strains reveals rapid genetic changes with clonal expansion during the late 1990s. J Bacteriol 186, 5496-5505.

van Amersfoorth, S.C., Schouls, L.M., van der Heide, H.G., Advani, A., Hallander, H.O., Bondeson, K., von Konig, C.H., Riffelmann, M., Vahrenholz, C., Guiso, N., Caro, V., Njamkepo, E., He, Q., Mertsola, J., Mooi, F.R., 2005. Analysis of Bordetella pertussis populations in European countries with different vaccination policies. J Clin Microbiol 43, 2837-2843.

van Gent, M., Heuvelman, C.J., van der Heide, H.G., Hallander, H.O., Advani, A., Guiso, N., Wirsing von Konig, C.H., Vestrheim, D.F., Dalby, T., Fry, N.K., Pierard, D., Detemmerman, L., Zavadilova, J., Fabianova, K., Logan, C., Habington, A., Byrne, M., Lutynska, A., Mosiej, E., Pelaz, C., Grondahl-Yli-Hannuksela, K., Barkoff, A.M., Mertsola, J., Economopoulou, A., He, Q., Mooi, F.R., 2015. Analysis of Bordetella pertussis clinical isolates circulating in European countries during the period 1998-2012. Eur J Clin Microbiol Infect Dis 34, 821-830.

van Loo, I.H., Heuvelman, K.J., King, A.J., Mooi, F.R., 2002. Multilocus sequence typing of Bordetella pertussis based on surface protein genes. J Clin Microbiol 40, 1994-2001.

World Health Organization, W., 2013. Pertussis.

World Health Organization, W., 2016. Immunization surveillance, assessment and monitoring.

Zhang, L., Xu, Y., Zhao, J., Kallonen, T., Cui, S., Xu, Y., Hou, Q., Li, F., Wang, J., He, Q., Zhang, S., 2010. Effect of vaccination on Bordetella pertussis strains, China. Emerg Infect Dis 16, 1695-1701.