Group A streptococci are important pathogens with various virulence factors, such as M protein, superantigens, hemolysins, deoxyribonuclease, and proteases. The aims of this study are to investigate the detection of emm genotypes and other virulence genes, such as SAgs, DNase, protease, antibiotic resistance, and phylogenetic relationships in GAS strains isolated from clinical samples.
Test strains were obtained from Çukurova University Balcalı Hospital and regional hospitals in Adana province. The M proteins were detected by sequence analysis of emm genes. SAgs and other virulence gene profiles were determined using the Multiplex-PCR method. The antibiotic susceptibility of the isolates was performed by the disc diffusion method and evaluated according to CLSI criteria. The PFGE method was used to determine the clonal relationship between the strains.
The emm gene was positive in 86 isolates. The most common emm genotypes were emm28 (22%), emm1 (18.6%), emm12 (13.9%), and emm3 (11.6%). Also, the most common virulence genes were speG (58.1%), speC (56.9%), sdaB (53.4%), and mac (53.4%). The rates of resistance to erythromycin, clindamycin, levofloxacin, ciprofloxacin and telithromycin were 19.8%, 16.3%, 4.7%, 3.5%, and 3.5%, respectively.
As a result, additional regional studies on the detection and prevalence of GAS virulence factors in Turkey are required. We believe that this study will provide valuable information for epidemiological studies on emm sequences, Sags, and other virulence factors of Streptococcus pyogenes in Turkey.
Steer AC, Lamagni T, Curtis N, Carapetis JR. Invasive group a streptococcal disease: epidemiology, pathogenesis and management. Drugs 2012; 72(9): 1213–1227.
Cunningham MW. Pathogenesis of group a streptococcal infection. Clin Microbiol Rev 2000; 13(3): 470–511.
Hasegawa T, Torii K, Hashikawa S, Iinuma Y, Ohta M. Cloning and characterization of two novel DNases from Streptococcus pyogenes. Arch Microbiol 2002; 177: 451–456.
Athey TB, Teatero S, Li A, Marchand-Austin A, Beall BW, Fittipaldi N. Deriving group a streptococcus typing ınformation from short-read whole-genome sequencing data. J Clin Microbiol 2014; 52(6): 1871–1876.
Pato C, Melo JC, Ramirez M. Friães A and the Portuguese Group for the Study of Streptococcal Infections. Streptococcus pyogenes causing skin and soft tissue ınfections are enriched in the recently emerged emm89 clade 3 and are not associated with abrogation of CovRS. Front Microbiol 2018; 9: 2372.
Ekelund K, Darenberg J, Teglund AN, Hoffmann S, Bang D, Skinhø P, et al. Variations in emm type among group a streptococcal isolates causing invasive or noninvasive infections in a nationwide study. J Clin Microbiol 2005; 43(7): 3101–3109.
CDC (Centers for Disease and Prevention). Available from: https://www.cdc.gov/streplab/groupa-strep/emm-background.html.
Khosravi AD, Ebrahimifard N, Shamsizadeh A, Shoja S. Isolation of Streptococcus pyogenes from children with pharyngitis and emm type analysis. J Chin Med Assoc 2016; 79: 276–280.
Friães A, Pinto FR, Silva-Costa C, Ramirez M, Melo-Cristino J. Portuguese Group for the Study of Streptococcal Infections. Group a streptococci clones associated with invasive infections and pharyngitis in Portugal present differences in emm types, superantigen gene content and antimicrobial resistance. BMC Microbiol 2012; 12: 280.
Imohl M, Fitzner C, Perniciaro S, Linden M. Epidemiologyand distribution of 10 superantigens among invasive Streptococcus pyogenes disease in Germany from 2009 to 2014. PLoS One 2017; 12(7): e0180757.
Commons RJ, Smeesters PR, Proft T, Fraser JD, Robins-Browne R, Curtis N. Streptococcal superantigens: categorization and clinical associations. Trends Mol Med 2014; 20 (1): 48–62.
Barnett TC, Cole JN, Hernandez TR, Henningham A, Paton JC, Nizet V, et al. Streptococcal toxins: role in pathogenesis and disease. Cell Microbiol 2015; 17(12): 1721–1741.
Dağı HT, Yüksekkaya Ş, Seyhan T, Fındık D, Tuncer İ, Arslan U. Investigation of Streptococcus pyogenes virulence factors and typing by multiple locus variable number tandem repeat fingerprinting (MLVF) method. Mikrobiyol Bul 2018; 52(3): 233–246.
Otlu B, Karakur C, Bayındır Y, Kayabaş Ü, Yakupoğulları Y, Bağ HG. Carriage of Streptococcus pyogenes in primary school children: M-protein types, pyrogenic toxin genes, and ınvestigation of the clonal relationships between the ısolates. Mikrobiyol Bul 2015; 49(3): 301–313.
CDC (Centers for Disease and Prevention). Available from: https://www.cdc.gov/streplab/groupa-strep/emm-typing-protocol.html.
Borek AL, Obszańska K, Hryniewicz W, Sitkiewicz I. Detection of Streptococcus pyogenes virulence factors by multiplex PCR. Virulence, Landes Biosci 2012; 3(6): 529–533.
CDC (Centers for Disease and Prevention). Available from: https://www.cdc.gov/pulsenet/pdf/listeria-pfge-protocol-508c.pdf.
Graves, LM, Swaminathan B. PulseNet standardized protocol for subtyping Listeria monocytogenes by macrorestriction and pulsed-field gel electrophoresis. Int J Food Microbiol 2001; 65(1–2): 55–62.
Karaky NM. Characterization of Streptococcus pyogenes ısolates from Lebanon: pathogenesis, epidemiology and resistance. Lebanese Amerıcan Unıversıty. A thesis Submitted in partial fulfillment of the requirements for the degree of Master of Science in Molecular Biology. Lebanon 2011. https://laur.lau.edu.lb:8443/xmlui/bitstream/handle/10725/5067/Nathalie_Mahmoud_Karaky_Thesis_Redacted.pdf?sequence=3.
Smit PW, Lindholm L, Lyytikainen O, Jalava J, Pätäri-Sampo A, Vuopio J. Epidemiology and emm types of invasive group a streptococcal infections in Finland, 2008-2013. Eur J Clin Microbiol Infect Dis 2015; 34(10): 2131–2136.
Meisal R, Andreasson IKG, Hooiby EA, Aaberge IS, Michaelsen TE, Caugant DA. Streptococcus pyogenes ısolates causing severe ınfections in Norway in 2006 to 2007: emm types, multilocus sequence types, and superantigen profiles. J Clin Microbiol 2010; 48(3): 842–851.
Akça G. Determination of M types of group a beta hemolytic streptococci by 'emm' sequencing analyse. PhD thesis. Ankara, 2006. Available from: https://tez.yok.gov.tr/UlusalTezMerkezi/giris.jsp.
Çakmak AE. M protein (emm) gene typing of group a beta-hemolytic streptococci isolated from pharyngeal cultures of children with acute tonsillopharyngitis in Istanbul. Child Health and Diseases Specialization Thesis. Istanbul, 2009. Available from: http://nek.istanbul.edu.tr:4444/ekos/TEZ/45289.pdf.
Dundar D, Sayan M, Tamer GS. Macrolide and tetracycline resistance and emm type distribution of Streptococcus pyogenes isolates recovered from Turkish patients. Microb Drug Resist 2010; 16(4): 279–284.
Arslan U, Oryaşın E, Eskin Z, Dağı HT, Fındık D, Tuncer İ, et al. Distribution of emm genotypes and antibiotic susceptibility of Streptococcus pyogenes strains: analogy with the vaccine in development. Mikrobiyol Bul 2013; 47(2): 318–323.
Strus M, Heczko PB, Golinska E, Tomusiak A, Chmielarczyk A, Dorycka M, et al. The virulence factors of group a streptococcus strains isolated from invasive and non-invasive infections in Polish and German centres, 2009–2011. Eur J Clin Microbiol Infect Dis 2017; 36(9): 1643–1649.
Friães A, Pinto FR, Silva-Costa C, Ramirez M, Melo-Cristino J. Superantigen gene complement of Streptococcus pyogenes relationship with other typing methods and short-term stability. Eur J Clin Microbiol Infect Dis 2013; 32(1): 115–125.
Mengeloğlu ZF. Evaluation of emm gene types, toxin gene profiles and clonal relatedness of group a streptococci obtained from patients and carriers. PhD thesis. Zonguldak, 2010. Available from: https://tez.yok.gov.tr/UlusalTezMerkezi/giris.jsp.
Karaky NM, Araj GF, Tokajian ST. Molecular characterization of Streptococcus pyogenes group a isolate from a tertiary hospital in Lebanon. J Med Microbiol 2014; 63(9): 1197–1204.