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Abstract

Purpose. Data on the clonal distribution of Staphylococcus aureus in Africa are scanty, partly due to the high costs and long turnaround times imposed by conventional genotyping methods such as spa and multilocus sequence typing (MLST), which means there is a need for alternative typing approaches. This study evaluated the discriminatory power, cost of and time required for genotyping Kenyan staphylococcal isolates using iPlex MassARRAY compared to conventional methods.

Methodology. Fifty-four clinical S. aureus isolates from three counties were characterized using iPlex MassARRAY, spa and MLST typing methods. Ten single-nucleotide polymorphisms (SNPs) from the S. aureus MLST loci were assessed by MassARRAY.

Results. The MassARRAY assay identified 14 unique SNP genotypes, while spa typing and MLST revealed 22 spa types and 21 sequence types (STs) that displayed unique regional distribution. spa type t355 (ST152) was the dominant type overall while t037/t2029 (ST 241) dominated among the methicillin-resistant S. aureus (MRSA) isolates. MassARRAY showed 83 % and 82 % accuracy against spa typing and MLST, respectively, in isolate classification. Moreover, MassARRAY identified all MRSA strains and a novel spa type. MassARRAY had a reduced turnaround time (<12 h) compared to spa typing (4 days) and MLST (20 days). The MassARRAY reagent and consumable costs per isolate were approximately $18 USD compared to spa typing ($30 USD) and MLST ($126 USD).

Conclusion. This study demonstrated that iPlex MassARRAY can be adapted as a useful surveillance tool to provide a faster, more affordable and fairly accurate method for genotyping African S. aureus isolates to identify clinically significant genotypes, MRSA strains and emerging strain types.

Keyword(s): Kenya , MassARRAY , MLST , S. aureus , spa and typing
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2019-05-03
2019-08-23
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References

  1. Abdulgader SM, Shittu AO, Nicol MP, Kaba M. Molecular epidemiology of Methicillin-resistant Staphylococcus aureus in Africa: a systematic review. Front Microbiol 2015;6:348 [CrossRef]
    [Google Scholar]
  2. DeLeo FR, Otto M, Kreiswirth BN, Chambers HF. Community-associated meticillin-resistant Staphylococcus aureus. The Lancet 2010;375:1557–1568 [CrossRef]
    [Google Scholar]
  3. Loughman JA, Fritz SA, Storch GA, Hunstad DA. Virulence gene expression in human community-acquired Staphylococcus aureus infection. J Infect Dis 2009;199:294–301 [CrossRef]
    [Google Scholar]
  4. Holmes A, McAllister G, McAdam PR, Hsien Choi S, Girvan K et al. Genome-wide single nucleotide polymorphism-based assay for high-resolution epidemiological analysis of the methicillin-resistant Staphylococcus aureus hospital clone EMRSA-15. Clin Microbiol Infect 2014;20:O124–O131 [CrossRef]
    [Google Scholar]
  5. Schaumburg F, Alabi AS, Peters G, Becker K. New epidemiology of Staphylococcus aureus infection in Africa. Clin Microbiol Infect 2014;20:589–596 [CrossRef]
    [Google Scholar]
  6. Omuse G, Van Zyl KN, Hoek K, Abdulgader S, Kariuki S et al. Molecular characterization of Staphylococcus aureus isolates from various healthcare institutions in Nairobi, Kenya: a cross sectional study. Ann Clin Microbiol Antimicrob 2016;15:51 [CrossRef]
    [Google Scholar]
  7. Akoru C, Kuremu RT, Ndege SK, Obala A, Smith JW et al. Prevalence and anti-microbial susceptibility of methicillin Staphylococcus aureus at Moi Teaching and Referral Hospital Eldoret. OJMM 2016;6:9–16 [CrossRef]
    [Google Scholar]
  8. Aiken AM, Mutuku IM, Sabat AJ, Akkerboom V, Mwangi J et al. Carriage of Staphylococcus aureus in Thika Level 5 Hospital, Kenya: a cross-sectional study. Antimicrob Resist Infect Control 2014;3:22 [CrossRef]
    [Google Scholar]
  9. Syrmis MW, Moser RJ, Whiley DM, Vaska V, Coombs GW et al. Comparison of a multiplexed MassARRAY system with real-time allele-specific PCR technology for genotyping of methicillin-resistant Staphylococcus aureus. Clin Microbiol Infect 2011;17:1804–1810 [CrossRef]
    [Google Scholar]
  10. Struelens MJ, Hawkey PM, French GL, Witte W, Tacconelli E. Laboratory tools and strategies for methicillin-resistant Staphylococcus aureus screening, surveillance and typing: state of the art and unmet needs. Clin Microbiol Infect 2009;15:112–119 [CrossRef]
    [Google Scholar]
  11. Kobayashi N, Wu H, Kojima K, Taniguchi K, Urasawa S et al. Detection of mecA, femA, and femB genes in clinical strains of staphylococci using polymerase chain reaction. Epidemiol Infect 1994;113:259–266 [CrossRef]
    [Google Scholar]
  12. Milheiriço C, Oliveira DC, de Lencastre H. Update to the multiplex PCR strategy for assignment of mec element types in Staphylococcus aureus. Antimicrob Agents Chemother 2007;51:3374–3377 [CrossRef]
    [Google Scholar]
  13. Shopsin B, Gomez M, Montgomery SO, Smith DH, Waddington M et al. Evaluation of protein A gene polymorphic region DNA sequencing for typing of Staphylococcus aureus strains. J Clin Microbiol 1999;37:3556–3563
    [Google Scholar]
  14. Enright MC, Day NP, Davies CE, Peacock SJ, Spratt BG. Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. J Clin Microbiol 2000;38:1008–1015
    [Google Scholar]
  15. Hall TA.BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT Nucleic acids symposium series London: Information Retrieval Ltd; 1999; ppc1979–c2000
    [Google Scholar]
  16. Harmsen D, Claus H, Witte W, Rothgänger J, Claus H et al. Typing of methicillin-resistant Staphylococcus aureus in a university hospital setting by using novel software for spa repeat determination and database management. J Clin Microbiol 2003;41:5442–5448 [CrossRef]
    [Google Scholar]
  17. Breurec S, Fall C, Pouillot R, Boisier P, Brisse S et al. Working group on Staphylococcus aureus infections, Garin B, Laurent F. Epidemiology of methicillin-susceptible Staphylococcus aureus lineages in five major African towns: high prevalence of Panton-Valentine leukocidin genes. Clin Microbiol Infect 2011;17:633–639
    [Google Scholar]
  18. Egyir B, Guardabassi L, Sørum M, Nielsen SS, Kolekang A et al. Molecular epidemiology and antimicrobial susceptibility of clinical Staphylococcus aureus from healthcare institutions in Ghana. PLoS One 2014;9:e89716 [CrossRef]
    [Google Scholar]
  19. Trembizki E, Smith H, Lahra MM, Chen M, Donovan B et al. High-throughput informative single nucleotide polymorphism-based typing of Neisseria gonorrhoeae using the Sequenom MassARRAY iPLEX platform. J Antimicrob Chemother 2014;69:1526–1532 [CrossRef]
    [Google Scholar]
  20. Whiley DM, Trembizki E, Buckley C, Freeman K, Baird RW et al. Molecular antimicrobial resistance surveillance for Neisseria gonorrhoeae, Northern Territory, Australia. Emerg Infect Dis 2017;23:1478–1485 [CrossRef]
    [Google Scholar]
  21. Robertson GA, Thiruvenkataswamy V, Shilling H, Price EP, Huygens F et al. Identification and interrogation of highly informative single nucleotide polymorphism sets defined by bacterial multilocus sequence typing databases. J Med Microbiol 2004;53:35–45 [CrossRef]
    [Google Scholar]
  22. Stephens AJ, Huygens F, Inman-Bamber J, Price EP, Nimmo GR et al. Methicillin-resistant Staphylococcus aureus genotyping using a small set of polymorphisms. J Med Microbiol 2006;55:43–51 [CrossRef]
    [Google Scholar]
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