The Presence of Mef (E) and Erm (B) Genes in Throat Samples of Children Infected with Streptococcus pyogenes

Year 2021, Volume: 11 Issue: 4, 445 - 451, 31.07.2021

Çiğdem Eda Balkan Bozlak Hayrunnisa Bekis Bozkurt Cem Öziç Ahmet Yılmaz

https://doi.org/10.16899/jcm.884444

Abstract

Aim: Antibiotic resistance studies about the Streptococcus have a great importance for the treatment of especially child throat infections in terms of public health and rational antibiotic use. For this purpose, we aimed to identify the macrolide resistance genes mef(E) and erm(B).
Material and Method: The throat culture samples taken from 51 children presenting to the hospital with the complaint of sore throat were evaluated in the laboratory, and S. pyogenes was diagnosed using tests; gram staining, catalase, bacitracin and PYR. Susceptibility profile was determined with the Kirby-Bauer disk-diffusion method and minimum inhibitor concentration(MICs) of erythromycin and clindamycin was determined by VITEK 2 System.
Results: S. pyogenes was possible macrolide resistance genes of mef(E) and erm(B) was determined by PCR. Macrolide resistance in S. pyogenes are determined in 51 pateients as follows; benzylpenicillin 0%, erytromycin 74.5%, clindamycin 31.4%. In patients with S. pyogenes, the genetic determinants of macrolide resistance mef(E) and erm(B) genes was investigated with the PCR method using primers specific to each gene. Total of 48% of the isolates (n=24) were positive for erm (B), 10% (n=5) for mef(E), in 42% (n=21) of the isolates both genes were detected.
Conclusion: The results of our study show that macrolid resistance genes were found in five of the patients evaluated. When the anamnesis of these patients was examined, it was determined that there were patients that frequently presented to the hospital with throat infections and experienced re-infection within a few weeks after receiving treatment.

Keywords

Moleculer epidemiology, mef(A) and Erm(B), Resistant genes, PCR, Streptecoccus pyogenes

Supporting Institution

Kafkas University

Project Number

2018-TS–67

Thanks

I think she is an expert in her field and will contribute to the development of the article

Streptococcus pyogenes ile Enfekte Çocukların Boğaz Örneklerinde Mef (E) ve Erm (B) Genlerinin Varlığı

Abstract

Amaç: Streptococcus ile ilgili yapılan antibiyotik direnç çalışmaları, özellikle çocuk boğaz enfeksiyonlarının tedavisinde halk sağlığı ve akılcı antibiyotik kullanımı açısından büyük önem taşımaktadır. Biz de çalışmamızda bu amaçla makrolid direnç genleri mef (E) ve erm (B) 'yi tanımlamayı amaçladık.
Yöntem ve Bulgular: Boğaz ağrısı şikayeti ile hastaneye başvuran 51 çocuktan alınan boğaz kültürü örnekleri laboratuvarda değerlendirildi ve gram boyama, katalaz, basitrasin ve PYR testleri ile S. pyogenes tanısı konuldu. Duyarlılık profili Kirby-Bauer disk difüzyon yöntemi ile ve Eritromisin ile klindamisinin minimum inhibitör konsantrasyonu (MİK), VITEK 2 Sistemi ile saptandı. S. pyogenes'te ki makrolid direnci 51 hastada aşağıdaki gibi saptandı; benzilpenisilin %0, eritromisin %74,5, klindamisin %31,4. S. pyogenes hastalarında makrolid direnci mef (E) ve erm (B) genlerinin genetik belirteçleri her gene özel primerler kullanılarak PCR yöntemi ile araştırıldı. İzolatların toplam% 48'i (n = 24) erm (B) için pozitif, % 10'u (n = 5) mef (E) için pozitif, %42'sinde (n = 21) her iki gen birlikte tespit edildi.
Sonuç: Çalışmamızın sonuçlarına göre makrolid direnç genleri pozitif hastaların anamnezleri incelendiğinde, hastaneye sık sık boğaz enfeksiyonu ile başvuran ve tedavi aldıktan sonraki birkaç hafta içinde tekrar enfeksiyon yaşayan hastaların olduğu belirlendi. S. pyogenes'te özellikle çocuklarda antimikrobiyal direnç tehdidi nedeniyle, direnci saptamak için ulusal ve şehir sürveyans çalışmalarına ihtiyaç vardır. Bu nedenle araştırmamız, geliştirmeye açık yeni hipotezler ve çalışmalarla desteklenmelidir.
Anahtar Kelimeler: Direnç genleri, PCR, Streptococcus pyogenes, Moleküler epidemiyoloji

Keywords

PCR, Direnç geni, moleküler epidemiyoloji

Project Number

2018-TS–67

References

• 1. Ferretti J, Köhler W. History of streptococcal research. In: Ferretti J, Stevens DL, Fischetti VA, editors. Streptococcus pyogenes: basic biology to clinical manifestations. Oklahoma: The University of Oklahoma Health Sciences Center; 2016.
• 2. Chu HY, Englund JA. Maternal immunization. Clin Infect Dis 2014;59(4): 560–8.
• 3. Othman AM, Assayaghi RM, Al-Shami HZ. Asymptomatic carriage of Streptococcus pyogenes among school children in Sana’a city, Yemen. BMC Research Notes 2019;12:339.
• 4. Steer AC, Carapetis JR, Dale JB et al. Status of research and development of vaccines for Streptococcus pyogenes. Vaccine 2016;34(26): 2953-58.
• 5. Musser JM, Beres SB, Zhu L et al. Reduced In Vitro Susceptibility of Streptococcus pyogenes to β-Lactam Antibiotics Associated with Mutations in the pbp2x Gene Is Geographically Widespread. J Clin Microbiol 2020;58(4): e01993-19.
• 6. Passali D, Lauriello, M, Passali, GC, Bellussi L. Group A streptococcus and its antibiotic resistance. Acta Otorhinolaryngol Ital 2007;27: 27–32.
• 7. Bley C, van der Linden M, Reinert RR. mef(A) is the predominant macrolide resistance determinant in Streptococcus pneumoniae and Streptococcus pyogenes in Germany. Int J Antimicrob Agents 2011;37(5): 425–31.
• 8. Topkaya AE, Balıkçı A, Aydın F et al. Epidemiology, clinical and microbiological characteristics of invasive streptococcal infections in Turkey, 2010-2011. Mikrobiyol Bul 2014;48(1):1-13. 9. Gröndal H. Harmless friendly and lethal: Antibiotic misuse in relation to the unpredictable bacterium Group A streptococcus. Sociology of Health & Illness 2018;40(7):1127–41.
• 10. Hansen SM, Uldbjerg N, Kilian M, Uffe B, Sørensen S. Dynamics of Streptococcus agalactiae colonization in women during and after pregnancy and in their infants. J Clin Microbiol 2004;42: 83–9.
• 11. Bauer AW, Kirby WMM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized disk method. Am J Clin Path 1966;45:493-6.
• 12. Gaertig J, Thatcher TH, Gu L, Gorovsky MA. Electroporation-mediated replacement of a positively and negatively selectable β-tubulin gene in Tetrahymena thermophile. Proc Natl Acad Sci 1994; 91(10): 4549-53. 13. Sağıroğlu P, Aksu B, Hasdemir MU. Investigation of macrolide resistance mechanisms in Streptococcus pneumoniae: results of Marmara University hospital between 2005–2008. Marmara Med J 2011;24:15–20.
• 14.https://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/Breakpoint_tables/v 10.0_Breakpoint_Tables.pdf
• 15. Hammond-Collins K, Strauss B, Barnes K et al. Group A streptococcus outbreak in a canadian armed forces training facility. Military medicine 2019;184(3-4):197-204.
• 16. Dewyer A, Scheel A, Webel AR, et al. Prevalence of group A β-hemolytic streptococcal throat carriage and prospective pilot surveillance of streptococcal sore throat in Ugandan school children. Int J Infect Dis 2020;93:245-51.
• 17. Hanage William P, Samuel A. Shelburne III. "Streptococcus pyogenes with reduced susceptibility to β-lactams: how big an alarm bell ?. " Clin Infect Dis 2020;71(1):205-6.
• 18. Kim KS, Kaplan EL. Association of penicillin tolerance with failure to eradicate group A streptococci from patients with pharyngitis. J Pediatr 1985;107(5):681-4.
• 19. Dicuonzo G, Fiscarelli E, Gherardi G et al. Erythromycin-resistant pharyngeal isolates of Streptococcus pyogenes recovered in Italy. Antimicrob Agents Chemother 2002;46:3987-90.
• 20. Dundar D, Sayan M, Tamer G. Macrolide and tetracycline resistance and emm type distribution of Streptococcus pyogenes isolates recovered from Turkish patients. Microbial Drug 2010;16(4):279-84.
• 21. Lu B, Fang Y, Fan Y, et al (2017). High Prevalence of Macrolide-resistance and Molecular Characterization of Streptococcus pyogenes Isolates Circulating in China from 2009 to 2016. Front Microbiol 2017;8(8):1052. 22. Bingen E, Fitoussi F, Doit C et al. Resistance to macrolides in Streptococcus pyogenes in France in pediatric patients. Antimicrob agents chemother 2000;44(6):1453-7.
• 23. Bozdogan B, Appelbaum PC, Kelly LM et al. Activity of telithromycin compared with seven other agentsagainst 1039 Streptococcus pyogenes pediatric isolates from ten centers in central and eastern. Europe Clin Microbiol Infect 2003;9(7):741-5.
• 24. Oryaşın E, Bıyık HH, Tristram S, Bozdoğan B. Cloned ermTR gene confers low level Erythromycin but high level Clindamycin resistance in Streptococcus pyogenes NZ131. Microbial Drug Resistance 2020;26(7):747-51.
• 25. Camara M, Dieng A, Boye C. Antibiotic susceptibility of Streptococcus pyogenes isolated from respiratory tract infections in Dakar, Senegal. Microbiol 2013;6:71–5.
• 26. Samir A, Abdel-Moein KA, Zaher HM. Emergence of penicillin-macrolide-resistant Streptococcus pyogenes among pet animals: An ongoing public health threat. Comp Immunol Microbiol Infect Dis 2020; 68:101390.
• 27. Dundar D, Sayan M, Tamer G. Macrolide and tetracycline resistance and emm type distribution of Streptococcus pyogenes isolates recovered from Turkishpatient. Microb Drug Resist 2010;16(4):279–84.
• 28. Bley C, van der Linden M, Reinert RR. mef(A) is the predominant macrolide resistance determinant in Streptococcus pneumoniae and Streptococcus pyogenes in Germany. J Antimicrob Agents 2011;37(5):425–31.
• 29. Vannice KS, Ricaldi J, Nanduri S et al. Streptococcus pyogenes pbp2x Mutation Confers Reduced Susceptibility to β-lactam antibiotics. Clin Infect Dis 2020;71(1): 201-4.
• 30. Öztop AY, Şanlıdağ T, Erandaç, M. Grouping and Determining of Antibiotic Susceptibilty of Beta-Haemolytic Streptococci Isolated from Children with Upper Respiratory Tract Infection. Turk Mikrobiyol Cem Derg 1999;30:73-6.
• 31. Sangvik M, Littauer P, Simonsen GS, Arnfinn S, Kristin HD. mef(A), mef(E) and a new mef allele in macrolide-resistant Streptococcus spp. İsolates from Norway, J Antimicrob Chemother 2005;56(5):841-6.
• 32. Littauer P, Caugant DA, Sangvik M, Høiby EA, Sundsfjord A, Simonsen GS. Macrolide-resistant Streptococcus pyogenes in Norway: population structure and resistance determinants, Antimicrob Agents Chemother 2006; 50(5):1896-99.
• 33. https://www.ecdc.europa.eu/en/search?s=cost+of+resistant+bacterias (accession date 14.01.2021).
• 34. Fitoussi F, Loukil C, Gros I et al. Mechanisms of macrolide resistance in clinical group B streptococci isolated in France. Antimicrob Agents Chemother 2001; 45:1889–91.