Skip to main content

Typing Candida Species Using Microsatellite Length Polymorphism and Multilocus Sequence Typing

  • Protocol
Candida Species

Abstract

To gain more insight into the epidemiological relationships between isolates of Candida spp. obtained from various origins, several molecular typing techniques have been developed. Two methods have emerged in the 2000s as soon as enough knowledge of the Candida spp. genomes was available to choose adequate loci and primers, namely microsatellite length polymorphism (MLP) and multilocus sequence typing (MLST). To contrast with previous PCR-based methods, specific amplifications with stringent conditions easily reproducible are the basis of MLP and MLST. MLST relies on Sanger sequencing to detect single-nucleotide polymorphisms within housekeeping genes. MLP needs a first in silico step to select tandemly repeated stretches of two to five nucleotides. One of the two primers used to amplify a microsatellite locus is labeled and fragment sizing is automatically performed using high-resolution electrophoresis platforms. MLST provides results easily comparable between laboratories and active MLST schemes are publicly available for the main Candida species. For comparative studies, MLP needs standards to compensate for the electrophoretic variations depending on the platforms used. Both methods can help us gain insight into the genetic relatedness of fungal isolates, both with advantages and drawbacks, and the choice of one method rather than the other depends on the task in question.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Hunter PR (1991) A critical review of typing methods for Candida albicans and their applications. Crit Rev Microbiol 17:417–434

    Article  CAS  PubMed  Google Scholar 

  2. Soll DR (2000) The ins and outs of DNA fingerprinting the infectious fungi. Clin Microbiol Rev 13:332–370

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  3. Graser Y, Volovsek M, Arrington J, Schonian G, Presber W, Mitchell TG, Vilgalys R (1996) Molecular markers reveal that population structure of the human pathogen Candida albicans exhibits both clonality and recombination. Proc Natl Acad Sci U S A 93:12473–12477

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  4. Tavanti A, Hensgens LA, Ghelardi E, Campa M, Senesi S (2007) Genotyping of Candida orthopsilosis clinical isolates by amplification fragment length polymorphism reveals genetic diversity among independent isolates and strain maintenance within patients. J Clin Microbiol 45:1455–1462

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  5. Gil-Lamaignere C, Roilides E, Hacker J, Muller FM (2003) Molecular typing for fungi—a critical review of the possibilities and limitations of currently and future methods. Clin Microbiol Infect 9:172–185

    Article  CAS  PubMed  Google Scholar 

  6. Bretagne S, Costa JM, Besmond C, Carsique R, Calderone R (1997) Microsatellite polymorphism in the promoter sequence of the elongation factor 3 gene of Candida albicans as the basis for a typing system. J Clin Microbiol 35:1777–1780

    PubMed Central  CAS  PubMed  Google Scholar 

  7. Botterel F, Desterke C, Costa C, Bretagne S (2001) Analysis of microsatellite markers of Candida albicans used for rapid typing. J Clin Microbiol 39:4076–4081

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  8. Bougnoux ME, Tavanti A, Bouchier C, Gow NA, Magnier A, Davidson AD, Maiden MC, D’Enfert C, Odds FC (2003) Collaborative consensus for optimized multilocus sequence typing of Candida albicans. J Clin Microbiol 41:5265–5266

    Google Scholar 

  9. Benson G (1999) Tandem repeats finder: a program to analyze DNA sequences. Nucleic Acids Res 27:573–580

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  10. Bougnoux ME, Morand S, d’Enfert C (2002) Usefulness of multilocus sequence typing for characterization of clinical isolates of Candida albicans. J Clin Microbiol 40:1290–1297

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Dodgson AR, Pujol C, Denning DW, Soll DR, Fox AJ (2003) Multilocus sequence typing of Candida glabrata reveals geographically enriched clades. J Clin Microbiol 41:5709–5717

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Bently DR, Dunham I (1995) Mapping human chromosomes. Curr Opin Genet Dev 5:328–334

    Article  CAS  PubMed  Google Scholar 

  13. Eloy O, Marque S, Botterel F, Stephan F, Costa JM, Lasserre V, Bretagne S (2006) Uniform distribution of three Candida albicans microsatellite markers in two French ICU populations supports a lack of nosocomial cross-contamination. BMC Infect Dis 6:162

    Article  PubMed Central  PubMed  Google Scholar 

  14. Stephan F, Bah MS, Desterke C, Rezaiguia-Delclaux S, Foulet F, Duvaldestin P, Bretagne S (2002) Molecular diversity and routes of colonization of Candida albicans in a surgical intensive care unit, as studied using microsatellite markers. Clin Infect Dis 35:1477–1483

    Article  PubMed  Google Scholar 

  15. Albano L, Bretagne S, Mamzer-Bruneel MF, Kacso I, Desnos-Ollivier M, Guerrini P, Le Luong T, Cassuto E, Dromer F, Lortholary O (2009) Evidence that graft-site candidiasis after kidney transplantation is acquired during organ recovery: a multicenter study in France. Clin Infect Dis 48:194–202

    Article  PubMed  Google Scholar 

  16. Sampaio P, Gusmao L, Correia A, Alves C, Rodrigues AG, Pina-Vaz C, Amorim A, Pais C (2005) New microsatellite multiplex PCR for Candida albicans strain typing reveals microevolutionary changes. J Clin Microbiol 43:3869–3876

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Foulet F, Nicolas N, Eloy O, Botterel F, Gantier JC, Costa JM, Bretagne S (2005) Microsatellite marker analysis as a typing system for Candida glabrata. J Clin Microbiol 43:4574–4579

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  18. Grenouillet F, Millon L, Bart JM, Roussel S, Biot I, Didier E, Ong AS, Piarroux R (2007) Multiple-locus variable-number tandem-repeat analysis for rapid typing of Candida glabrata. J Clin Microbiol 45:3781–3784

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Desnos-Ollivier M, Bretagne S, Bernede C, Robert V, Raoux D, Chachaty E, Forget E, Lacroix C, Dromer F (2008) Clonal population of flucytosine-resistant Candida tropicalis from blood cultures, Paris, France. Emerg Infect Dis 14:557–565

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  20. Lasker BA, Butler G, Lott TJ (2006) Molecular genotyping of Candida parapsilosis group I clinical isolates by analysis of polymorphic microsatellite markers. J Clin Microbiol 44:750–759

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  21. Sabino R, Sampaio P, Rosado L, Stevens DA, Clemons KV, Pais C (2010) New polymorphic microsatellite markers able to distinguish among Candida parapsilosis sensu stricto isolates. J Clin Microbiol 48:1677–1682

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  22. Garcia-Hermoso D, Cabaret O, Lecellier G, Desnos-Ollivier M, Hoinard D, Raoux D, Costa JM, Dromer F, Bretagne S (2007) Comparison of microsatellite length polymorphism and multilocus sequence typing for DNA-based typing of Candida albicans. J Clin Microbiol 45:3958–3963

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. de Valk HA, Meis JF, Klaassen CH (2007) Microsatellite based typing of Aspergillus fumigatus: strengths, pitfalls and solutions. J Microbiol Methods 69:268–272

    Article  PubMed  Google Scholar 

  24. Pasqualotto AC, Denning DW, Anderson MJ (2007) A cautionary tale: lack of consistency in allele sizes between two laboratories for a published multilocus microsatellite typing system. J Clin Microbiol 45:522–528

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  25. Garcia-Hermoso D, MacCallum DM, Lott TJ, Sampaio P, Serna MJ, Grenouillet F, Klaassen CH, Bretagne S (2010) Multicenter collaborative study for standardization of Candida albicans genotyping using a polymorphic microsatellite marker. J Clin Microbiol 48:2578–2581

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  26. Costa JM, Garcia-Hermoso D, Olivi M, Cabaret O, Farrugia C, Lecellier G, Dromer F, Bretagne S (2010) Genotyping of Candida albicans using length fragment and high-resolution melting analyses together with minisequencing of a polymorphic microsatellite locus. J Microbiol Methods 80:306–309

    Article  CAS  PubMed  Google Scholar 

  27. Enright MC, Spratt BG (1999) Multilocus sequence typing. Trends Microbiol 7:482–487

    Article  CAS  PubMed  Google Scholar 

  28. Tavanti A, Davidson AD, Johnson EM, Maiden MC, Shaw DJ, Gow NA, Odds FC (2005) Multilocus sequence typing for differentiation of strains of Candida tropicalis. J Clin Microbiol 43:5593–5600

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Jacobsen MD, Gow NA, Maiden MC, Shaw DJ, Odds FC (2007) Strain typing and determination of population structure of Candida krusei by multilocus sequence typing. J Clin Microbiol 45:317–323

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  30. Daniel HM, Lachance MA, Kurtzman CP (2014) On the reclassification of species assigned to Candida and other anamorphic ascomycetous yeast genera based on phylogenetic circumscription. Antonie Van Leeuwenhoek 106:67–84

    Article  PubMed  Google Scholar 

  31. Lachance MA, Boekhout T, Scorzetti G, Fell JW, Kurtzman CP (2011) Candida Berkhout (1923). In: Kurtzman CP, Fell JW, Boekhout T (eds) The Yeasts, a taxonomic study. Elsevier, Amsterdam, pp 987–1278

    Chapter  Google Scholar 

  32. Donnelly SM, Sullivan DJ, Shanley DB, Coleman DC (1999) Phylogenetic analysis and rapid identification of Candida dubliniensis based on analysis of ACT1 intron and exon sequences. Microbiology 145(Pt 8):1871–1882

    Article  CAS  PubMed  Google Scholar 

  33. Costa JM, Eloy O, Botterel F, Janbon G, Bretagne S (2005) Use of microsatellite markers and gene dosage to quantify gene copy numbers in Candida albicans. J Clin Microbiol 43:1387–1389

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  34. Jolley KA, Chan MS, Maiden MC (2004) Mlstdbnet—distributed multi-locus sequence typing (MLST) databases. BMC Bioinformatics 5:86

    Article  PubMed Central  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Stéphane Bretagne M.D., Ph.D. .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Garcia-Hermoso, D., Desnos-Ollivier, M., Bretagne, S. (2016). Typing Candida Species Using Microsatellite Length Polymorphism and Multilocus Sequence Typing. In: Calderone, R., Cihlar, R. (eds) Candida Species. Methods in Molecular Biology, vol 1356. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3052-4_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3052-4_15

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3051-7

  • Online ISBN: 978-1-4939-3052-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics