Abstract
Streptococcus pyogenes encodes multiple virulence factors and their presence is often related to the severity of the disease. We designed the system of four low-volume multiplex PCR reactions to detect genes encoding 20 virulence factors: spd3, sdc, sdaB, sdaD, speB, spyCEP, scpA, mac, sic, speL, speK, speM, speC, speI, speA, speH, speG, speJ, smeZ, and ssa. Classification of strains based on the virulence factors absence or presence correlates with PFGE MLST and emm typing results. The typing/detection system is fast and cost-effective, can be used to detect GAS virulence factors and as a rapid tool to effectively differentiate between strains.
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References
Sitkiewicz I (2018) How to become a killer, or is it all accidental? Virulence strategies in oral streptococci. Mol Oral Microbiol 33:1–12
Sitkiewicz I, Hryniewicz W (2010) Pyogenic streptococci—danger of re-emerging pathogens. Pol J Microbiol 59:219–226
Carapetis JR, Steer AC, Mulholland EK et al (2005) The global burden of group a streptococcal diseases. Lancet Infect Dis 5:685–694
Sims Sanyahumbi A, Colquhoun S, Wyber R et al (2016) Global disease burden of group a streptococcus. In: Ferretti JJ, Stevens DL, Fischetti VA (eds) Streptococcus pyogenes: basic biology to clinical manifestations. University of Oklahoma Health Sciences Center, Oklahoma City, OK
Hauser AR, Stevens DL, Kaplan EL et al (1991) Molecular analysis of pyrogenic exotoxins from Streptococcus pyogenes isolates associated with toxic shock-like syndrome. J Clin Microbiol 29:1562–1567
Unnikrishnan M, Altmann DM, Proft T et al (2002) The bacterial superantigen streptococcal mitogenic exotoxin Z is the major immunoactive agent of Streptococcus pyogenes. J Immunol 169:2561–2569
Beres SB, Musser JM (2007) Contribution of exogenous genetic elements to the group a streptococcus metagenome. PLoS One 2:e800
Brouwer S, Barnett TC, Rivera-Hernandez T et al (2016) Streptococcus pyogenes adhesion and colonization. FEBS Lett 590:3739–3757
Rohde M, Cleary PP (2016) Adhesion and invasion of streptococcus pyogenes into host cells and clinical relevance of intracellular streptococci. In: Ferretti JJ, Stevens DL, Fischetti VA (eds) Streptococcus pyogenes: basic biology to clinical manifestations [Internet]. University of Oklahoma Health Sciences Center, Oklahoma City, OK. PMID: 26866223, NBK333420
Ryan PA, Juncosa B (2016) Group a streptococcal adherence. In: Ferretti JJ, Stevens DL, Fischetti VA (eds) Streptococcus pyogenes: basic biology to clinical manifestations. University of Oklahoma Health Sciences Center, Oklahoma City, OK
Fischetti VA (2016) M protein and other surface proteins on streptococci. In: Ferretti JJ, Stevens DL, Fischetti VA (eds) Streptococcus pyogenes: basic biology to clinical manifestations. University of Oklahoma Health Sciences Center, Oklahoma City, OK
Beall B.W. Protocol for emm typing. https://www.cdc.gov/streplab/groupa-strep/emm-typing-protocol.html. Accessed 08 Jul 2019
Proft T, Fraser JD (2016) Streptococcal Superantigens: biological properties and potential role in disease. In: Ferretti JJ, Stevens DL, Fischetti VA (eds) Streptococcus pyogenes: basic biology to clinical manifestations. University of Oklahoma Health Sciences Center, Oklahoma City, OK
Hynes W, Sloan M (2016) Secreted extracellular virulence factors. In: Ferretti JJ, Stevens DL, Fischetti VA (eds) Streptococcus pyogenes: basic biology to clinical manifestations. University of Oklahoma Health Sciences Center, Oklahoma City, OK
Walker MJ, Barnett TC, McArthur JD et al (2014) Disease manifestations and pathogenic mechanisms of group a streptococcus. Clin Microbiol Rev 27:264–301
Llewelyn M, Cohen J (2002) Superantigens: microbial agents that corrupt immunity. Lancet Infect Dis 2:156–162
Commons RJ, Smeesters PR, Proft T et al (2013) Streptococcal superantigens: Categorization and clinical associations. Trends Mol Med 20:1–15
Reglinski M, Sriskandan S, Turner CE (2019) Identification of two new core chromosome-encoded superantigens in Streptococcus pyogenes; speQ and speR. J Infect 78(5):358–363
Sumby P, Porcella SF, Barbian KD et al (2005) Evolutionary origin and emergence of a highly successful clone of serotype M1 group a streptococcus involved multiple horizontal gene transfer events. J Infect Dis 192:771–782
Walker MJ, Hollands A, Sanderson-Smith ML et al (2007) DNase Sda1 provides selection pressure for a switch to invasive group a streptococcal infection. Nat Med 13:981–985
Chang A, Khemlan A, Kang H et al (2011) Functional analysis of Streptococcus pyogenes nuclease a (SpnA), a novel group a streptococcal virulence factor. Mol Microbiol 79:1629–1642
de Buhr N, Neumann A, Jerjomiceva N et al (2014) Streptococcus suis DNase SsnA contributes to degradation of neutrophil extracellular traps (NETs) and evasion of NET-mediated antimicrobial activity. Microbiology 160:385–395
Ly AT, Noto JP, Walwyn OL et al (2017) Differences in SpeB protease activity among group a streptococci associated with superficial, invasive, and autoimmune disease. PLoS One 12:e0177784
Olsen RJ, Raghuram A, Cantu C et al (2015) The majority of 9,729 group a streptococcus strains causing disease secrete SpeB cysteine protease: pathogenesis implications. Infect Immun 83:4750–4758
Chiang-Ni C, Wu J-JJ (2008) Effects of streptococcal pyrogenic exotoxin B on pathogenesis of Streptococcus pyogenes. J Formos Med Assoc 107:677–685
Chen CC, Cleary PP (1990) Complete nucleotide sequence of the streptococcal C5a peptidase gene of Streptococcus pyogenes. J Biol Chem 265:3161–3167
Brown CK, Gu Z, Matsuka Y et al (2005) Structure of the streptococcal cell wall C5a peptidase. Proc Natl Acad Sci U S A 102:18391–18396
Pawel-Rammingen U, Björck L (2003) IdeS and SpeB: immunoglobulin-degrading cysteine proteinases of Streptococcus pyogenes. Curr Opin Microbiol 6:50–55
Lei B, DeLeo FR, Hoe NP et al (2001) Evasion of human innate and acquired immunity by a bacterial homolog of CD11b that inhibits opsonophagocytosis. Nat Med 7:1298–1305
Okumura CYM, Anderson EL, Döhrmann S et al (2013) IgG protease mac/IdeS is not essential for phagocyte resistance or mouse virulence of M1T1 group a streptococcus. MBio 4(4):e00499–e00413
Kurupati P, Turner CE, Tziona I et al (2010) Chemokine-cleaving Streptococcus pyogenes protease SpyCEP is necessary and sufficient for bacterial dissemination within soft tissues and the respiratory tract. Mol Microbiol 76:1387–1397
Hidalgo-Grass C, Mishalian I, Dan-Goor M et al (2006) A streptococcal protease that degrades CXC chemokines and impairs bacterial clearance from infected tissues. EMBO J 25:4628–4637
Borek AL, Wilemska J, Izdebski R et al (2011) A new rapid and cost-effective method for detection of Phages , ICEs and virulence factors encoded by Streptococcus pyogenes. Pol J Microbiol 60:187–201
Lintges M, Arlt S, Uciechowski P et al (2007) A new closed-tube multiplex real-time PCR to detect eleven superantigens of Streptococcus pyogenes identifies a strain without superantigen activity. Int J Med Microbiol 297:471–478
Acknowledgments
The work was supported by grants 2017/27/B/NZ7/00040 and 2018/29/B/NZ6/00624 to I.S.
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Kozińska, A., Sitkiewicz, I. (2020). Detection of Streptococcus pyogenes Virulence Factors. In: Proft, T., Loh, J. (eds) Group A Streptococcus. Methods in Molecular Biology, vol 2136. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0467-0_1
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DOI: https://doi.org/10.1007/978-1-0716-0467-0_1
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