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Chapter 8.8 : Full Identification of Yeasts

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Abstract:

In an era of increasing resistance of yeast species to antifungal agents and a widening range of species capable of causing diseases previously the domain of , there is almost no situation in which identification to species level is not warranted. This is especially true given the growth in the number of immune compromised patients in our society, which has provided more opportunities for yeast infections to occur and to complicate and prolong the recovery period. Molecular methods to identify yeasts directly in specimens and after growth in culture are under development. Since the last edition of this handbook, peptide nucleic acid fluorescence in situ hybridization (PNAFISH) technology (AdvanDx, Woburn, MA) has been developed, which allows identification of several yeasts directly from blood cultures. However, no commercially available method which identifies a broad range of species has been introduced. In this procedure, methods which take advantage of physiological characteristics of various yeast species are presented. Molecular methods are discussed in general terms.

Citation: Garcia L. 2010. Full Identification of Yeasts, p 442-455. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch8.8
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References

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1. Atlas, R. M. 1993. Handbook of Microbiological Media. CRC Press, Boca Raton, FL..
2. Barnett, J. A.,, R. W. Payne,, and D. Yarrow. 2000. Yeasts: Characteristics and Identification, 3rd ed. Cambridge University Press, Cambridge, United Kingdom.
3. Boekhout, T.,, and R. W. Bosboom. 1994. Karyotyping of Malassezia yeasts: taxonomic and epidemiological implications. Syst. Appl. Microbiol. 17: 146 153.
4. Burgener-Kairuz, P.,, J.-P. Zuber,, P. Jaunin,, T. G. Buchman,, J. Bille,, and M. Rossier. 1994. Rapid detection and identification of Candida albicans and Torulopsis (Candida) glabrata in clinical specimens by species-specific nested PCR amplification of a cytochrome P-450 lanosterol-α-demethylase (L1A1) gene fragment. J. Clin. Microbiol. 32: 1902 1907.
5. Campa, D.,, A. Tavanti,, F. Gemignani,, C. S. Mogavero,, I. Bellini,, F. Bottari,, R. Barale,, S. Landi,, and S. Senesi. 2008. DNA microarray based on arrayed-primer extension technique for identification of pathogenic fungi responsible for invasive and superficial mycoses. J. Clin. Microbiol. 46: 909 915.
6. Carlotti, A.,, F. Chaib,, A. Couble,, N. Bourgeois,, V. Blanchard,, and J. Villard. 1997. Rapid identification and fingerprinting of Candida krusei by PCR-based amplification of the species-specific repetitive polymorphic sequence CKRS-1. J. Clin. Microbiol. 35: 1337 1343.
7. Chen, Y. C.,, J. D. Eisner,, M. M. Kattar,, S. L. Rassoulian-Barrett,, K. LaFe,, S. L. Yarfitz,, A. P. Limaye,, and B. T. Cookson. 2000. Identification of medically important yeasts using PCR-based detection of DNA sequence polymorphisms in the internal transcribed spacer 2 region of the rRNA genes. J. Clin. Microbiol. 38: 2302 2310.
8. Clemons, K. V.,, F. Feroze,, K. Holmberg,, and D. A. Stevens. 1997. Comparative analysis of genetic variability among Candida albicans isolates from different geographic locales by three genotypic methods. J. Clin. Microbiol. 35: 1332 1336.
9. De Baere, T.,, G. Claeys,, D. Swinne,, G. Verschraegen,, A. Muylaert,, C. Massonet,, and M. Vaneechoutte. 2002. Identification of cultured isolates of clinically important yeast species using fluorescent fragment length analysis of the amplified internally transcribed rRNA spacer 2 region (ITS2). BMC Microbiol. 2: 21.
10. Defontaine, A.,, M. Coarer,, and J. P. Bouchara. 1996. Contribution of various techniques of molecular analysis to strain identification of Candida glabrata. Microbiol. Ecol. Health Dis. 9: 27 33.
11. Elie, C. M.,, T. J. Lott,, E. Reiss,, and C. J. Morrison. 1998. Rapid identification of Candida species with species-specific DNA probes. J. Clin. Microbiol. 36: 3260 3265.
12. Enger, L.,, S. Joly,, C. Pujol,, P. Simonson,, M. A. Pfaller,, and D. A. Soll. 2001. Cloning and characterization of a complex DNA fingerprinting probe for Candida parapsilosis. J. Clin. Microbiol. 39: 658 669.
13. Freydiere, A.-M.,, R. Guinet,, and P. Boiron. 2001. Yeast identification in the clinical microbiology laboratory: phenotypical methods. Med. Mycol. 39: 9 33.
14. Fujita, S.-I.,, and T. Hashimoto. 2000. DNA fingerprinting patterns of Candida species using HinfI endonuclease. Int. J. Syst. Evol. Microbiol. 50: 1381 1389.
15. Gaitanis, G.,, A. Velegraki,, E. Frangoulis,, A. Mitroussia,, A. Tsigonia,, A. Tzimogianni,, A. Katsambas,, and N. J. Legakis. 2002. Identification of Malassezia species from patient skin scales by PCR-RFLP. Clin. Microbiol. Infect. 8: 162 173.
16. Guého, E.,, L. Improvisi,, G. S. de Hoog,, and B. Dupont. 1994. Trichosporon on humans: a practical account. Mycoses 37: 3 10.
17. Gueho, E.,, G. Midgley,, and J. Guillot. 1996. The genus Malassezia with description of four new species. Antonie van Leeuwenhoek 69: 337 355.
18. Guillot, J.,, E. Guého,, M. Lesourd,, G. Midgley,, G. Chévrier,, and B. Dupont. 1996. Identification of Malassezia species. J. Mycol. Med. 6: 103 110.
19. Hazen, K. C. 1995. New and emerging yeast pathogens. Clin. Microbiol. Rev. 8: 462 478.
20. Hazen, K. C.,, and S. A. Howell,. 2003. Candida, Cryptococcus, and other yeasts of medical importance, p. 1693 1711. In P. R. Murray,, E. J. Baron,, K. Chapin,, M. A. Pfaller,, and F. C. Tenover (ed.), Manual of Clinical Microbiology, 8th ed. ASM Press, Washington, DC.
21. Hirai, A.,, R. Kano,, K. Makimura,, E. R. Duarte,, J. S. Hamdan,, M. A. Lachance,, H. Yamaguchi,, and A. Hasegawa. 2004. Malassezia nana sp. nov., a novel lipid-dependent yeast species isolated from animals. Int. J. Syst. Evol. Microbiol. 54: 623 627.
22. Howell, S. A.,, C. Quin,, and G. Midgley. 1993. Karyotypes of oval cell forms of Malassezia furfur. Mycoses 36: 263 266.
23. Jaeger, E. M. E.,, N. M. Carrol,, S. Choudhury,, A. A. S. Dunlop,, H. M. A. Towler,, M. M. Matheson,, P. Adamson,, N. Okhravi,, and S. Lightman. 2000. Rapid detection and identification of Candida, Aspergillus, and Fusarium species in ocular samples using nested PCR. J. Clin. Microbiol. 38: 2902 2908.
24. Joly, S.,, C. Pujol,, K. Schröppel,, and D. R. Soll. 1996. Development of two species-specific fingerprinting probes for broad computer assisted epidemiological studies of Candida tropicalis J. Clin. Microbiol. 34: 3063 3071.
25. Kwon-Chung, K. J.,, I. Polacheck,, and J. E. Bennett. 1982. Improved diagnostic medium for separation of Cryptococcus neoformans var. neoformans (serotypes A and D) and Cryptococcus neoformans var. gattii (serotypes B and C). J. Clin. Microbiol. 15: 535 537.
26. Landlinger, C.,, L. Baskova,, S. Preuner,, B. Willinger,, V. Buchta,, and T. Lion. 23 December 2008. Identification of fungal species by fragment length analysis of the internally transcribed spacer 2 region. Eur. J. Clin. Microbiol. Infect. Dis. doi: 10.1007/s10096-008-0683-3.
27. Larone, D. H. 2002. Medically Important Fungi: a Guide to Identification, 4th ed. ASM Press, Washington, DC.
28. Latouche, G. N.,, H.-M. Daniel,, O. C. Lee,, T. G. Mitchell,, T. C. Sorrell,, and W. Meyer. 1997. Comparison of use of phenotypic and genotypic characteristics for identification of species of the anamorph genus Candida and related teleomorph yeast species. J. Clin. Microbiol. 35: 3171 3180.
29. Linton, C. J.,, A. M. Borman,, G. Cheung,, A. D. Holmes,, A. Szekely,, M. D. Palmer,, P. D. Bridge,, C. K. Campbell,, and E. M. Johnson. 2007. Molecular identification of unusual pathogenic yeast isolates by large ribosomal subunit gene sequencing: 2 years of experience at the United Kingdom Mycology Reference Laboratory. J. Clin. Microbiol. 45: 1152 1158.
30. Lockhart, S. R.,, S. Joly,, C. Pujol,, J. D. Sobel,, M. A. Pfaller,, and D. R. Soll. 1997. Development and verification of fingerprinting probes for Candida glabrata. Microbiology 143: 3733 3746.
31. Magee, B. B.,, T. M. D'Souza,, and P. T. Magee. 1987. Strain and species identification by restriction fragment length polymorphisms in the ribosomal DNA repeat of Candida species. J. Bacteriol. 169: 1639 1643.
32. Mannarelli, B. M.,, and C. P. Kurtzman. 1998. Rapid identification of Candida albicans and other human pathogenic yeasts by using short oligonucleotides in a PCR. J. Clin. Microbiol. 36: 1634 1641.
33. Pincus, D. H.,, D. C. Coleman,, W. R. Pruitt,, A. A. Padhye,, I. F. Salkin,, M. Geimer,, A. Bassel,, D. J. Sullivan,, M. Clarke,, and V. Hearn. 1999. Rapid identification of Candida dubliniensis with commercial yeast identification systems. J. Clin. Microbiol. 37: 3533 3539.
34. Posteraro, B.,, M. Sanguinetti,, L. Masucci,, L. Romano,, G. Morace,, and G. Fadda. 2000. Reverse cross hybridization assay for rapid detection of PCR-amplified DNA from Candida species, Cryptococcus neoformans, and Saccharomyces cerevisiae in clinical samples. J. Clin. Microbiol. 38: 1609 1614.
35. Roy, B.,, and S. A. Meyer. 1998. Confirmation of the distinct genotype groups within the form species Candida parapsilosis. J. Clin. Microbiol. 36: 216 218.
36. Scherer, S.,, and D. A. Stevens. 1988. A Candida albicans dispersed, repeated gene family and its epidemiologic applications. Proc. Natl. Acad. Sci. USA 85: 1452 1456.
37. Schmid, J.,, E. Voss,, and D. R. Soll. 1990. Computer-assisted methods for assessing strain relatedness in Candida albicans by fingerprinting with the moderately repetitive sequence Ca3. J. Clin. Microbiol. 28: 1236 1243.
38. Shadomy, H. J.,, S. Wood-Helie,, S. Shadomy,, W. E. Dismukes,, R. Y. Chau, and the NIAID Mycoses Study Group. 1987. Biochemical serogrouping of clinical isolates of Cryptococcus neoformans. Diagn. Microbiol. Infect. Dis. 6: 131 138.
39. Shin, J. H.,, F. S. Nolte,, and C. J. Morrison. 1997. Rapid identification of Candida species in blood cultures by a clinically useful PCR method. J. Clin. Microbiol. 35: 1454 1459.
40. Soll, D. A. 2000. The ins and outs of DNA fingerprinting the infectious fungi. Clin. Microbiol. Rev. 13: 332 370.
41. Sugita, T.,, M. Tajima,, M. Takashima,, M. Amaya,, M. Saito,, R. Tsuboi,, and A. Nishikawa. 2004. A new yeast, Malassezia yamatoensis, isolated from a patient with seborrheic dermatitis, and its distribution in patients and healthy subjects. Microbiol. Immunol. 48: 579 583.
42. Sugita, T.,, M. Takashima,, M. Kodama,, R. Tsuboi,, and A. Nishikawa. 2003. Description of a new yeast species, Malassezia japonica, and its detection in patients with atopic dermatitis and healthy subjects. J. Clin. Microbiol. 41: 4695 4699.
43. Sugita, T.,, M. Takashima,, T. Shinoda,, H. Suto,, T. Unno,, R. Tsuboi,, H. Ogawa,, and A. Nishikawa. 2002. New yeast species, Malassezia dermatis, isolated from patients with atopic dermatitis. J. Clin. Microbiol. 40: 1363 1367.
44. Sullivan, D. J.,, T. J. Westerneng,, K. A. Haynes,, D. E. Bennett,, and D. C. Coleman. 1995. Candida dubliniensis sp. nov.: phenotypic and molecular characterization of a novel species associated with oral candidosis in HIV-infected individuals. Microbiology 141: 1507 1521.
45. Tavanti, A.,, A. D. Davidson,, N. A. Gow,, M. C. J. Maiden,, and F. C. Odds. 2005. Candida orthopsilosis and Candida metapsilosis spp. nov. to replace Candida parapsilosis groups II and III. J. Clin. Microbiol. 43: 284 292.
46. Thanos, M.,, G. Schönian,, W. Meyer,, C. Schweynoch,, Y. Gräser,, T. G. Mitchell,, W. Presber,, and H.-J. Tietz. 1996. Rapid identification of Candida species by DNA fingerprinting with PCR. J. Clin. Microbiol. 34: 615 621.
47. Wahyuningshi, R.,, H.-J. Freisleben,, H.-G. Sonntag,, and P. Schnitzler. 2000. Simple and rapid detection of Candida albicans DNA in serum by PCR for diagnosis of invasive candidiasis. J. Clin. Microbiol. 38: 3016 3021.
48. Warren, N. G.,, and K. C. Hazen,. 1999. Candida, Cryptococcus, and other yeasts of medical importance, p. 1184 1199. In P. R. Murray,, E. J. Baron,, M. A. Pfaller,, F. C. Tenover,, and R. H. Yolken (ed.), Manual of Clinical Microbiology, 7th ed. ASM Press, Washington, DC.
49. White, T. C.,, T. Bruns,, S. Lee,, and J. Taylor,. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics, p. 315 322. In M. A. Innis,, D. H. Gelfand,, J. J. Sninsky,, and T. C. White (ed.), PCR Protocols. A Guide to Methods and Applications. Academic Press, San Diego, CA.
50. Wickerham, L. J.,, and K. A. Burton. 1948. Carbon assimilation tests for the classification of yeasts. J. Bacteriol. 56: 363 371.
51. Williams, D. W.,, M. J. Wilson,, M. A. O. Lewis,, and A. J. C. Potts. 1995. Identification of Candida species by PCR and restriction fragment length polymorphism analysis of intergenic spacer regions of ribosomal DNA. J. Clin. Microbiol. 33: 2476 2479.
52. Wise, M. G.,, M. Healy,, K. Reece,, R. Smith,, D. Walton,, W. Dutch,, A. Renwick,, J. Huong,, S. Young,, J. Tarrand,, and D. P. Kontoyiannis. 2007. Species identification and strain differentiation of clinical Candida isolates using the DiversiLab system of automated repetitive sequence-based PCR. J. Med. Microbiol. 56: 778 787.
53. Wolk, D. M.,, and G. D. Roberts,. 2002. Commercial methods for identification and susceptibility testing of fungi, p. 225 255. In A. L. Truant (ed.), Manual of Commercial Methods in Clinical Microbiology. ASM Press, Washington, DC.
54. Remel. 1993. Technical information, ascospore agar. TI no. 9046-A. Remel, Lenexa, KS.
1. Snyder, J. W.,, and R. H. Atlas. 2006. Handbook of Media for Clinical Microbiology, 2nd ed. CRC Press, Boca Raton, FL..

Tables

Generic image for table
Table 8.8-1

Anamorph-teleomorph binomials of commonly encountered yeasts

Modified from reference .

Citation: Garcia L. 2010. Full Identification of Yeasts, p 442-455. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch8.8
Generic image for table
Table 8.8-2

General considerations of two commercial yeast identification systems

Citation: Garcia L. 2010. Full Identification of Yeasts, p 442-455. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch8.8
Generic image for table
Table 8.8-3

Examples of useful supplemental tests for yeasts

Note that some tests are components of commercial systems. They are listed because they are key tests.

+, positive; −, negative.

Fermentation tests require that the yeast first be grown on a sugar-free medium, such as malt extract agar or yeast morphology agar.

Citation: Garcia L. 2010. Full Identification of Yeasts, p 442-455. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch8.8
Generic image for table
Table 8.8-4

Culture and biochemical characteristics of yeasts frequently isolated from clinical specimens

Modified from Warren and Hazen ( ) and Pincus et al. ( ). +, positive; −, negative; *, some isolates may give the opposite reaction; R, rare; F, the sugar is fermented (i.e., gas was produced); W, weak fermentation.

+ growth greater than that of the negative control.

typically produces single and no more than two terminal chlamydospores, while some isolates of will produce terminal chlamydospore in pairs, triplets, and clusters.

assimilates erythritol; does not. Maximum growth temperatures, 43 to 45°C for and 33 to 37°C for .

assimilates rhamnose; usually does not.

assimilates L-arabinose; usually does not.

is a thermophilic yeast capable of growth at 40 to 42°C.

Rare strains of produce teardrop-shaped chlamydospores.

Not yeasts but may be confused with several yeast genera.

Citation: Garcia L. 2010. Full Identification of Yeasts, p 442-455. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch8.8
Generic image for table
Table 8.8-5

Characteristics of selected spp.

Modified from Hazen and Howell ( ) and Guého et al. ( ).

The carbohydrates are available from numerous chemical suppliers, including Sigma Chemical Co. (St. Louis, MO).

+, positive; −, negative; V, strain variation.

Also called an infection peg, which penetrates host tissue (hair).

Citation: Garcia L. 2010. Full Identification of Yeasts, p 442-455. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch8.8
Generic image for table
Table 8.8-6

Fermentation reactions for some Candida spp.

Note that and produce distinctively different morphologies on morphology media. and are physiologically very similar. The difficulty in distinguishing these organisms is that no single differential test (e.g., growth at 45°C, color formation on CHROMagar, or assimilation profile) is definitive. However, several (at least three) differential tests in combination provide sufficient information to identify an organism as one or the other species. Otherwise, molecular techniques should be employed to distinguish the species.

−, negative; +, positive; o, not tested routinely; v, rare negatives; V, strain variation; w, weak.

Citation: Garcia L. 2010. Full Identification of Yeasts, p 442-455. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch8.8
Generic image for table
Table 8.8-7

Key characteristics to differentiate species

All species except require lipid supplementation for growth. +, positive; −, negative.

This test is performed as described in references and .

w, weak. This test is performed by placing a drop of HO (10%) either directly on the plate culture or onto a smear of the culture on a glass slide ( ).

Growth of is inhibited by high concentrations of Tween 20. At lower concentrations, the organism exhibits Tween 20 assimilation ( ).

and can be distinguished by their morphologies. is globose, whereas produces long, cylindrical cells.

Citation: Garcia L. 2010. Full Identification of Yeasts, p 442-455. In Clinical Microbiology Procedures Handbook, 3rd Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817435.ch8.8

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