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Journal of Clinical Microbiology, February 2004, p. 639-644, Vol. 42, No. 2
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.2.639-644.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Genetic Diversity of Cell-Invasive Erythromycin-Resistant and -Susceptible Group A Streptococci Determined by Analysis of the RD2 Region of the prtF1 Gene

Cinzia Spinaci,1 Gloria Magi,1 Claudia Zampaloni,2 Luca A. Vitali,2 Claudia Paoletti,1 Maria R. Catania,3 Manuela Prenna,2 Luigi Ferrante,1 Sandro Ripa,2 Pietro E. Varaldo,1 and Bruna Facinelli1*

Department of Microbiology and Biomedical Sciences, University of Ancona Medical School, 60131 Ancona ,1 Department of Molecular Cellular Animal Biology, University of Camerino, 62032 Camerino ,2 Department of Experimental Medicine, University of Naples Medical School, 80138 Naples, Italy3

Received 21 July 2003/ Returned for modification 20 September 2003/ Accepted 13 November 2003

The RD2 region of the internalization-associated gene prtF1, which encodes the fibronectin-binding repeat domain type 2 of protein F1, plays a crucial role in the entry of group A streptococci (GAS) into epithelial cells. A molecular study of the variability of the RD2 region was carried out with 77 independent Italian GAS, 66 erythromycin resistant (ER) and 11 erythromycin susceptible (ES), which had previously been investigated for the association between erythromycin resistance and ability to enter human respiratory cells. The amplicons obtained from PCR analysis of the RD2 region were consistent with a number of RD2 repeats ranging from one to five, more frequently four (n = 30), three (n = 27), and one (n = 18). A new method to type cell-invasive GAS (RD2 typing) was developed by combining PCR analysis of the RD2 region and restriction analysis of PCR products with endonucleases HaeIII, DdeI, and HinfI. Overall, 10 RD2 types (a to j) were distinguished (all detected among the 66 ER isolates, four detected among the 11 ES isolates). Comparison and correlation of RD2 typing data with the genotype and phenotype of macrolide resistance and with data from PCR M typing and SmaI macrorestriction analysis allowed us to identify 41 different clones (31 among the 66 ER isolates and 10 among the 11 ES isolates). Three major clones accounted for 40% of the isolates (47% of ER strains). Some ES isolates appeared to be related to ER isolates with identical combinations of RD2 type and emm type. While simultaneous use of different typing methods is essential for a thorough investigation of GAS epidemiology, RD2 typing may be especially helpful in typing cell-invasive GAS.

* Corresponding author. Mailing address: Department of Microbiology and Biomedical Sciences, University of Ancona Medical School, Via Ranieri, Monte d'Ago 60131 Ancona, Italy. Phone: 39 071 2204695. Fax: 071 2204693. E-mail: b.facinelli{at}univpm.it.

Journal of Clinical Microbiology, February 2004, p. 639-644, Vol. 42, No. 2
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.2.639-644.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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