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. 1995 Nov;33(11):2818–2822. doi: 10.1128/jcm.33.11.2818-2822.1995

Genetic relatedness of Cryptococcus neoformans clinical isolates grouped with the repetitive DNA probe CNRE-1.

F Chen 1, B P Currie 1, L C Chen 1, S G Spitzer 1, E D Spitzer 1, A Casadevall 1
PMCID: PMC228586  PMID: 8576325

Abstract

Cryptococcus neoformans isolates from eight patients with cryptococcal infection were previously assigned into three groups on the basis of repetitive DNA probe (CNRE-1) restriction fragment length polymorphisms. These groups accounted for a disproportionate number of recent clinical isolates in New York City. To further examine the genetic relatedness of isolates within and across CNRE-1 groups, the DNA sequence of the 779-base URA5 gene from each strain was amplified and sequenced. The number of nucleotide differences occurred in the third codon position or in introns. Pairwise comparisons revealed average nucleotide differences within a CNRE-1 group of 4.8 +/- 2.6 (n = 8) and between CNRE-1 groups of 21.9 +/- 7.0 (n =20) (P <0.001) Analysis of URA5 sequences defined three groups that were congruent with those defined by CNRE-1 restriction fragment length polymorphisms. PCR amplification of an rDNA intergenic spacer revealed conservation of the intergenic spacer length within groups. Electrophoretic karyotyping did not distinguish between two isolates in each of two CNRE-1 groups. DNA from all isolates studied hybridized to an alpha mating type-specific probe. We interpret these results as suggesting a clonal population structure for some pathogenic isolates of C. neoformans in New York City.

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Selected References

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  1. Casadevall A., Freundlich L. F., Marsh L., Scharff M. D. Extensive allelic variation in Cryptococcus neoformans. J Clin Microbiol. 1992 May;30(5):1080–1084. doi: 10.1128/jcm.30.5.1080-1084.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Currie B. P., Casadevall A. Estimation of the prevalence of cryptococcal infection among patients infected with the human immunodeficiency virus in New York City. Clin Infect Dis. 1994 Dec;19(6):1029–1033. doi: 10.1093/clinids/19.6.1029. [DOI] [PubMed] [Google Scholar]
  3. Currie B. P., Freundlich L. F., Casadevall A. Restriction fragment length polymorphism analysis of Cryptococcus neoformans isolates from environmental (pigeon excreta) and clinical sources in New York City. J Clin Microbiol. 1994 May;32(5):1188–1192. doi: 10.1128/jcm.32.5.1188-1192.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Currie B. P., Freundlich L. F., Soto M. A., Casadevall A. False-negative cerebrospinal fluid cryptococcal latex agglutination tests for patients with culture-positive cryptococcal meningitis. J Clin Microbiol. 1993 Sep;31(9):2519–2522. doi: 10.1128/jcm.31.9.2519-2522.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Edman J. C., Kwon-Chung K. J. Isolation of the URA5 gene from Cryptococcus neoformans var. neoformans and its use as a selective marker for transformation. Mol Cell Biol. 1990 Sep;10(9):4538–4544. doi: 10.1128/mcb.10.9.4538. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fan M., Chen L. C., Ragan M. A., Gutell R. R., Warner J. R., Currie B. P., Casadevall A. The 5S rRNA and the rRNA intergenic spacer of the two varieties of Cryptococcus neoformans. J Med Vet Mycol. 1995 Jul-Aug;33(4):215–221. doi: 10.1080/02681219580000451. [DOI] [PubMed] [Google Scholar]
  7. Kwon-Chung K. J., Bennett J. E. Distribution of alpha and alpha mating types of Cryptococcus neoformans among natural and clinical isolates. Am J Epidemiol. 1978 Oct;108(4):337–340. doi: 10.1093/oxfordjournals.aje.a112628. [DOI] [PubMed] [Google Scholar]
  8. Mitchell T. G., Freedman E. Z., White T. J., Taylor J. W. Unique oligonucleotide primers in PCR for identification of Cryptococcus neoformans. J Clin Microbiol. 1994 Jan;32(1):253–255. doi: 10.1128/jcm.32.1.253-255.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Moore T. D., Edman J. C. The alpha-mating type locus of Cryptococcus neoformans contains a peptide pheromone gene. Mol Cell Biol. 1993 Mar;13(3):1962–1970. doi: 10.1128/mcb.13.3.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Perfect J. R., Magee B. B., Magee P. T. Separation of chromosomes of Cryptococcus neoformans by pulsed field gel electrophoresis. Infect Immun. 1989 Sep;57(9):2624–2627. doi: 10.1128/iai.57.9.2624-2627.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Spitzer E. D., Spitzer S. G., Freundlich L. F., Casadevall A. Persistence of initial infection in recurrent Cryptococcus neoformans meningitis. Lancet. 1993 Mar 6;341(8845):595–596. doi: 10.1016/0140-6736(93)90354-j. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Spitzer S. G., Spitzer E. D. Characterization of the CNRE-1 family of repetitive DNA elements in Cryptococcus neoformans. Gene. 1994 Jun 24;144(1):103–106. doi: 10.1016/0378-1119(94)90211-9. [DOI] [PubMed] [Google Scholar]
  15. Tibayrenc M., Kjellberg F., Arnaud J., Oury B., Brenière S. F., Dardé M. L., Ayala F. J. Are eukaryotic microorganisms clonal or sexual? A population genetics vantage. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5129–5133. doi: 10.1073/pnas.88.12.5129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Varma A., Kwon-Chung K. J. DNA probe for strain typing of Cryptococcus neoformans. J Clin Microbiol. 1992 Nov;30(11):2960–2967. doi: 10.1128/jcm.30.11.2960-2967.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Wickes B. L., Moore T. D., Kwon-Chung K. J. Comparison of the electrophoretic karyotypes and chromosomal location of ten genes in the two varieties of Cryptococcus neoformans. Microbiology. 1994 Mar;140(Pt 3):543–550. doi: 10.1099/00221287-140-3-543. [DOI] [PubMed] [Google Scholar]

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