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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Jul;33(7):1890–1895. doi: 10.1128/jcm.33.7.1890-1895.1995

Comparison of multilocus enzyme electrophoresis and random amplified polymorphic DNA analysis for molecular subtyping of Cryptococcus neoformans. The Cryplococcal Disease Active Surveillance Group.

M E Brandt 1, L C Hutwagner 1, R J Kuykendall 1, R W Pinner 1
PMCID: PMC228292  PMID: 7665665

Abstract

We evaluated multilocus enzyme electrophoresis (MEE) and random amplified polymorphic DNA (RAPD) for their usefulness in subtyping 344 Cryptococcus neoformans clinical isolates obtained from four U.S. metropolitan areas in 1992 to 1994. MEE and RAPD with five primers both discriminated between the two varieties of C. neofromans. MEE divided C. neoformans var. neoformans isolates into 15 enzyme electrophoretic subtypes (ETs) arranged in three complexes. The predominant ET 1 complex contained 10 ETs, with isolates from 70% of patients in 1 ET. RAPD with five primers further sorted this predominant ET into 19 subtypes, with 60% of isolates sorting into three RAPD types. The ET 8 MEE complex, containing three ETs, could not be divided further by RAPD. The ET 7 complex (two ETs) included isolates from all serotype AD patients. Although both MEE and RAPD identified isolates of C. neoformans var. gattii, neither distinguished between serotypes B and C. These results showed that the two C. neoformans varieties could be identified by MEE or RAPD profile as well as by biochemical methods. RAPD improved the discriminatory power of MEE for isolates within the ET 1 complex but with other ETs offered little additional sensitivity over MEE and was less sensitive than MEE with isolates of C. neoformans var. gattii. This information will be useful in identifying particular environmental sources of disease-causing exposures, in seeking clusters of cases, and in determining whether an infecting strain changes over time.

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Selected References

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  1. Bennett J. E., Kwon-Chung K. J., Howard D. H. Epidemiologic differences among serotypes of Cryptococcus neoformans. Am J Epidemiol. 1977 Jun;105(6):582–586. doi: 10.1093/oxfordjournals.aje.a112423. [DOI] [PubMed] [Google Scholar]
  2. Brandt M. E., Bragg S. L., Pinner R. W. Multilocus enzyme typing of Cryptococcus neoformans. J Clin Microbiol. 1993 Oct;31(10):2819–2823. doi: 10.1128/jcm.31.10.2819-2823.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Böttger E. C. Frequent contamination of Taq polymerase with DNA. Clin Chem. 1990 Jun;36(6):1258–1259. [PubMed] [Google Scholar]
  4. Cherniak R., Sundstrom J. B. Polysaccharide antigens of the capsule of Cryptococcus neoformans. Infect Immun. 1994 May;62(5):1507–1512. doi: 10.1128/iai.62.5.1507-1512.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dykstra M. A., Friedman L., Murphy J. W. Capsule size of Cryptococcus neoformans: control and relationship to virulence. Infect Immun. 1977 Apr;16(1):129–135. doi: 10.1128/iai.16.1.129-135.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ellsworth D. L., Rittenhouse K. D., Honeycutt R. L. Artifactual variation in randomly amplified polymorphic DNA banding patterns. Biotechniques. 1993 Feb;14(2):214–217. [PubMed] [Google Scholar]
  7. Grant I. H., Armstrong D. Fungal infections in AIDS. Cryptococcosis. Infect Dis Clin North Am. 1988 Jun;2(2):457–464. [PubMed] [Google Scholar]
  8. Ikeda R., Shinoda T., Fukazawa Y., Kaufman L. Antigenic characterization of Cryptococcus neoformans serotypes and its application to serotyping of clinical isolates. J Clin Microbiol. 1982 Jul;16(1):22–29. doi: 10.1128/jcm.16.1.22-29.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kaplan W., Bragg S. L., Crane S., Ahearn D. G. Serotyping Cryptococcus neoformans by immunofluorescence. J Clin Microbiol. 1981 Sep;14(3):313–317. doi: 10.1128/jcm.14.3.313-317.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kwon-Chung K. J., Bennett J. E. Epidemiologic differences between the two varieties of Cryptococcus neoformans. Am J Epidemiol. 1984 Jul;120(1):123–130. doi: 10.1093/oxfordjournals.aje.a113861. [DOI] [PubMed] [Google Scholar]
  11. Kwon-Chung K. J., Polacheck I., Bennett J. E. 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. 1982 Mar;15(3):535–537. doi: 10.1128/jcm.15.3.535-537.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lehmann P. F., Kemker B. J., Hsiao C. B., Dev S. Isoenzyme biotypes of Candida species. J Clin Microbiol. 1989 Nov;27(11):2514–2521. doi: 10.1128/jcm.27.11.2514-2521.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lehmann P. F., Lin D., Lasker B. A. Genotypic identification and characterization of species and strains within the genus Candida by using random amplified polymorphic DNA. J Clin Microbiol. 1992 Dec;30(12):3249–3254. doi: 10.1128/jcm.30.12.3249-3254.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. MacPherson J. M., Eckstein P. E., Scoles G. J., Gajadhar A. A. Variability of the random amplified polymorphic DNA assay among thermal cyclers, and effects of primer and DNA concentration. Mol Cell Probes. 1993 Aug;7(4):293–299. doi: 10.1006/mcpr.1993.1043. [DOI] [PubMed] [Google Scholar]
  15. Meyer W., Mitchell T. G., Freedman E. Z., Vilgalys R. Hybridization probes for conventional DNA fingerprinting used as single primers in the polymerase chain reaction to distinguish strains of Cryptococcus neoformans. J Clin Microbiol. 1993 Sep;31(9):2274–2280. doi: 10.1128/jcm.31.9.2274-2280.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Möller E. M., Bahnweg G., Sandermann H., Geiger H. H. A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi, fruit bodies, and infected plant tissues. Nucleic Acids Res. 1992 Nov 25;20(22):6115–6116. doi: 10.1093/nar/20.22.6115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nei M. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics. 1978 Jul;89(3):583–590. doi: 10.1093/genetics/89.3.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Panther L. A., Sande M. A. Cryptococcal meningitis in the acquired immunodeficiency syndrome. Semin Respir Infect. 1990 Jun;5(2):138–145. [PubMed] [Google Scholar]
  19. Polacheck I., Lebens G., Hicks J. B. Development of DNA probes for early diagnosis and epidemiological study of cryptococcosis in AIDS patients. J Clin Microbiol. 1992 Apr;30(4):925–930. doi: 10.1128/jcm.30.4.925-930.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Selander R. K., Caugant D. A., Ochman H., Musser J. M., Gilmour M. N., Whittam T. S. Methods of multilocus enzyme electrophoresis for bacterial population genetics and systematics. Appl Environ Microbiol. 1986 May;51(5):873–884. doi: 10.1128/aem.51.5.873-884.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Selander R. K., McKinney R. M., Whittam T. S., Bibb W. F., Brenner D. J., Nolte F. S., Pattison P. E. Genetic structure of populations of Legionella pneumophila. J Bacteriol. 1985 Sep;163(3):1021–1037. doi: 10.1128/jb.163.3.1021-1037.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Selander R. K., Musser J. M., Caugant D. A., Gilmour M. N., Whittam T. S. Population genetics of pathogenic bacteria. Microb Pathog. 1987 Jul;3(1):1–7. doi: 10.1016/0882-4010(87)90032-5. [DOI] [PubMed] [Google Scholar]
  23. Shimizu R. Y., Howard D. H., Clancy M. N. The variety of Cryptococcus neoformans in patients with AIDS. J Infect Dis. 1986 Dec;154(6):1042–1042. doi: 10.1093/infdis/154.6.1042. [DOI] [PubMed] [Google Scholar]
  24. Spitzer E. D., Spitzer S. G. Use of a dispersed repetitive DNA element to distinguish clinical isolates of Cryptococcus neoformans. J Clin Microbiol. 1992 May;30(5):1094–1097. doi: 10.1128/jcm.30.5.1094-1097.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sugar A. M. Overview: cryptococcosis in the patient with AIDS. Mycopathologia. 1991 Jun;114(3):153–157. doi: 10.1007/BF00437205. [DOI] [PubMed] [Google Scholar]
  26. Tibayrenc M., Neubauer K., Barnabé C., Guerrini F., Skarecky D., Ayala F. J. Genetic characterization of six parasitic protozoa: parity between random-primer DNA typing and multilocus enzyme electrophoresis. Proc Natl Acad Sci U S A. 1993 Feb 15;90(4):1335–1339. doi: 10.1073/pnas.90.4.1335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Todaro-Luck F., Reiss E., Cherniak R., Kaufman L. Characterization of Cryptococcus neoformans capsular glucuronoxylomannan polysaccharide with monoclonal antibodies. Infect Immun. 1989 Dec;57(12):3882–3887. doi: 10.1128/iai.57.12.3882-3887.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Varma A., Kwon-Chung K. J. Restriction fragment polymorphism in mitochondrial DNA of Cryptococcus neoformans. J Gen Microbiol. 1989 Dec;135(12):3353–3362. doi: 10.1099/00221287-135-12-3353. [DOI] [PubMed] [Google Scholar]
  29. Wang G., Whittam T. S., Berg C. M., Berg D. E. RAPD (arbitrary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains. Nucleic Acids Res. 1993 Dec 25;21(25):5930–5933. doi: 10.1093/nar/21.25.5930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Welsh J., McClelland M. Genomic fingerprinting using arbitrarily primed PCR and a matrix of pairwise combinations of primers. Nucleic Acids Res. 1991 Oct 11;19(19):5275–5279. doi: 10.1093/nar/19.19.5275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Williams J. G., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. doi: 10.1093/nar/18.22.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]

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