Skip to main content
Log in

Characterization ofCryptococcus neoformans var.neoformans serotype A and A/D in samples from Egypt

  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

The cryptococcal polysaccharide antigen was detected in 10 cerebrospinal fluid (CSF) and 23 serum samples from cryptococcal meningitis and intestinal cryptococcosis by the cryptococcal antigen latex agglutination system (CALAS). CALAS titers in CSF and serum samples of cryptococcal meningitis ranged over 8–2048 and 32–2048, respectively, while in cases of intestinal cryptococcosis, serum titers ranged over 8-2048. The isolates of yeastCryptococcus neoformans were determined to be of serotype A or of the A/D pair. The total leukocyte count and biochemical parameters in CSF were significantly increased as indicators of microbial infection. Furthermore, thein vitro change of the teleomorph (sexual state) to the anamorph (asexual state) was also detected and the teleomorph state changedin vivo to the encapsulated anamoph state which is more virulent during infectionin vivo than the yeast-like noncapsulated form. Two primers for internal transcribed spacer (ITS) regions of ribosomal DNA were used for molecular detection ofC. neoformans. After PCR amplification, a DNA band of 415 bp, visualized on agarose gel, indicated the presence ofC. neoformans cells in the tested CSF and serum samples. The primer sensitivity was also characterized using purified yeast chromosomal DNA as template; it was about 20 pg or more chromosomal DNA which represents about 10 cells ofC. neoformans. The primers were also specific for ITS regions ofC. neoformans and gave negative results withCandida albicans andE. coli chromosomal DNA templates.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Abdel-Fatah A., Abuzeid A.: Primary cutaneous cryptococcosis in Egypt.Internat.J.Dermatol. 14, 606–608 (1976).

    Article  Google Scholar 

  • Arnaoot H., Mostafa W., Ekladios E.: Disseminated cryptococosis with cutaneous lesions.5th Internat. Conf. Microbiology, AIDS and Emerging Infectious Diseases, Cairo (Egypt) (1996).

  • Atlas R.M., Parks L.C.:Handbook of Microbiological Media, 2nd ed. CRC Press, Boca Raton-New York-London-Tokyo 1997.

    Google Scholar 

  • Casadevall A., Freundlich L., Marsh L., Scharff M.D.: Extensive allelic variation inC. neoformans.J.Clin.Microbiol. 30, 1080–1084 (1992).

    PubMed  CAS  Google Scholar 

  • Chang H.C., Leaw S.N., Huang A.H., Wu T.L., Chang T.C.: Rapid identification of yeasts in positive blood cultures by a multiplex PCR method.J.Clin.Microbiol. 39, 3466–3471 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Chen F., Currie B.P., Chen L., Spitzer S., Spitzer E., Casadevall A.: Genetic relatedness ofC. neoformans clinical isolates grouped with the repetitive DNA probe.J.Clin.Microbiol. 33, 2818–2822 (1995).

    PubMed  CAS  Google Scholar 

  • Currie B., Freundlich L., Casadevall A.: Restriction fragment length polymorphism analysis ofCryptococcus neoformans isolates from environmental (pigeon excreta) and clinical sources in New York City.J.Clin.Microbiol. 32, 1188–1192 (1994).

    PubMed  CAS  Google Scholar 

  • Dorko E., Kmeťová M., Pilipčinec E., Bračoková I., Dorko F., Danko J., Švicky E., Tkáčikova L’.: Rare non-albicans Candida species detected in different clinical diagnoses.Folia Microbiol. 45, 364–368 (2000a).

    Article  CAS  Google Scholar 

  • Dorko E., Kmeťová M., Dorko F., Bračoková I., Danko J., Švický E., Tkáčiková L’.: Prevalence ofCryptococcus neoformans in clinical specimens.Folia Microbiol. 45, 369–374 (2000b).

    Article  CAS  Google Scholar 

  • Dorko E., Kmeťová M., Pilipčinec E., Dorko F., Bračokova I., Marossy A., Škardová I.: Non-albicans Candida species in cancer patients.The Sciences 1, 11–15 (2001a).

    Google Scholar 

  • Dorko E., Pilipčinec E., Mahel’ M., Virágová S., Bračoková I., Dorko F., Švicky E., Danko J., Holoda E., Ondrašović M., Tkáčikova L’.: Yeast-like microorganisms in eye infections.Folia Microbiol. 46, 147–150 (2001b).

    Article  CAS  Google Scholar 

  • Dorko E., Pilipčinec E., Tkáčiková L’.: Candidal urinary tract infections caused by non-albicans Candida species.Folia Microbiol. 47, 182–184 (2002a).

    Article  CAS  Google Scholar 

  • Dorko E., Pilipčinec E., Tkačiková L’.: Fungal disease of the respiratory tract.Folia Microbiol. 47, 302–304 (2002b).

    Article  CAS  Google Scholar 

  • Dromer F., Gueho E., Ronin O., Dupont B.: Serotyping ofC. neoformans by using a monoclonal antibody specific for capsular polysaccharide.J.Clin.Microbiol. 31, 359–363 (1993).

    PubMed  CAS  Google Scholar 

  • Dromer F., Varma A., Ronin O., Mathoulin S., Dupont B.: Molecular typing ofC. neoformans serotype D clinical isolates.J.Clin. Microbiol. 32, 2364–2371 (1994).

    PubMed  CAS  Google Scholar 

  • Ellis D., Pfeiffer T.: The ecology ofCryptococcus neoformans.Eur.J.Epidemiol. 8, 321–325 (1992).

    Article  PubMed  CAS  Google Scholar 

  • Fessel W.J.: Cryptococcal meningitis after unusual exposure to birds.N.Engl.J.Med. 328, 1354–1355 (1993).

    Article  PubMed  CAS  Google Scholar 

  • Franzot S., Fries B., Cleare W., Casadevall A.: Genetic relationship betweenC. neoformans var.neoformans strains of serotypes A and D.J.Clin.Microbiol. 36, 2200–2204 (1998).

    PubMed  CAS  Google Scholar 

  • Fromtling R., Abruzzo G., Ruiz A.: Virulence and antifungal susceptibility of environmental and clinical isolates ofCryptococcus neoformans from Puerto Rico.Mycopathologia 106, 163–166 (1989).

    Article  PubMed  CAS  Google Scholar 

  • Hosney H.Y., Refai M.K.:Fungi and the Lung: Treatise on Research. Mycology Unit, Chest Department, Ain-Shams University, Cairo (Egypt) 1986.

    Google Scholar 

  • Ikeda R., Shinoda T., Fukazawa Y., Kaufman L.: Antigenic characterization ofC. neoformans serotypes and its application to serotyping of clinical isolates.J.Clin.Microbiol. 16, 22–29 (1982).

    PubMed  CAS  Google Scholar 

  • Ikeda R., Sugita T., Shinoda T.: Serological relationship ofCryptococcus spp.: distribution of antigenic factors inCryptococcus and interspecies diversity.J.Clin.Microbiol. 38, 4021–4025 (2000).

    PubMed  CAS  Google Scholar 

  • Kwon-Chung K., Polacheck K., Popkin T.: Improved diagnostic medium for separation ofC. neoformans var.neoformans (serotype A and D) andC. neoformans var.gattii (serotype B and C).J.Clin.Microbiol. 15, 335–337 (1982).

    Google Scholar 

  • Kwon-Chung K., Wicker B., Stockman L., Roberts G., Ellis D., Howard D.: Virulence, serotype and molecular characteristics of environmental strains ofCryptococcus neoformans var.gattii.Infect.Immun. 60, 1869–1874 (1992).

    PubMed  CAS  Google Scholar 

  • Levitz S.M.: The ecology ofCryptococcus neoformans and the epidemiology of cryptococcosis.Rev.Infect.Dis. 13, 1163–1169 (1991).

    PubMed  CAS  Google Scholar 

  • Magee J.T., Philpot C., Yang J., Hosein I.K.: Pyrolysis typing of isolates from a recurrence of systemic cryptococcosis.J.Med. Microbiol. 40, 165–169 (1994).

    Article  PubMed  CAS  Google Scholar 

  • Mitchell T., Freedman E., White T., Taylor J.: Unique oligonucleotide primers in PCR for identification ofC. neoformans.J.Clin. Microbiol. 32, 253–255 (1994).

    PubMed  CAS  Google Scholar 

  • Nakamura Y., Kano R., Watanabe S., Hasegawa A.: Molecular analysis ofCAP59 gene-sequencing from five serotypes ofC. neoformans.J.Clin.Microbiol. 38, 992–995 (2000).

    PubMed  CAS  Google Scholar 

  • Perfect J.R.: Cryptococcosis.Infect.Dis.Clin.N.Am. 3, 77–102 (1989).

    CAS  Google Scholar 

  • Perfect J.R., Magee B.B., Magee P.T.: Separation of chromosomes ofC. neoformans by pulsed field gel electrophoresis.Infect. Immun. 57, 2624–2627 (1989).

    PubMed  CAS  Google Scholar 

  • Rappelli P., Are R., Casu G., Fiori P.L., Cappuccinelli P., Aceti A.: Development of a nested PCR for detection ofC. neoformans in cerebrospinal fluid.J.Clin.Microbiol. 36, 3438–3440 (1998).

    PubMed  CAS  Google Scholar 

  • Ruiz A., Formtling R.A., Bulmer G.S.: Distribution ofCryptococcus neoformans in a natural site.Infect.Immun. 31, 560–563 (1981).

    PubMed  CAS  Google Scholar 

  • Salkin I., Hurd N.: New medium for differentiation ofCryptococcus neoformans serotype pairs.J.Clin.Microbiol. 15, 169–171 (1982).

    PubMed  CAS  Google Scholar 

  • Sandhu G.S., Kline B., Stockman L., Roberts G.: Molecular probes for diagnosis of fungal infections.J.Clin.Microbiol. 33, 2913–2919 (1995).

    PubMed  CAS  Google Scholar 

  • Steenbergen N., Casadevall A.: Prevalence ofCryptococcus neoformans var.neoformans (serotype D) andCryptococcus neoformans var.grubii (serotype A) isolates in New York City.J.Clin.Microbiol. 38, 1974–1976 (2000).

    PubMed  CAS  Google Scholar 

  • Swinne D., Kayembe K., Niyimi M.: Isolation saprophyticCryptococcus neoformans var.neoformans in Kinshasa, Zaire.Ann.Soc. Belg.Med.Trop. 66, 57–61 (1986).

    PubMed  CAS  Google Scholar 

  • Tanaka K., Miyazaki T., Maesaki T., Mitsutake K., Kakeya H., Yamamoto Y., Yanagihara K., Hossain M.A., Tashiro T., Kohno S.: Detection ofC. neoformans gene with pulmonary cryptococcosis.J.Clin.Microbiol. 34, 2826–2828 (1996).

    PubMed  CAS  Google Scholar 

  • Temstet A., Roux P., Poirot J., Ronin O., Dramer F.: Evaluation of a monoclonal antibody-based latex agglutination test for diagnosis of cryptococcosis: comparison with two tests using polyclonal antibodies.J.Clin.Microbiol. 30, 2544–2550 (1992).

    PubMed  CAS  Google Scholar 

  • Tomšikova A.: Causative agents of nosocomial mycoses.Folia Microbiol. 47, 105–112 (2002).

    Article  Google Scholar 

  • Varma A., Kwon-Chung K.: DNA probe for strain typing ofCryptococcus neoformans.J.Clin.Microbiol. 30, 2960–2967 (1992).

    PubMed  CAS  Google Scholar 

  • Varma A., Swinne D., Staib F., Bennett J.E., Kwon-Chung J.E.: Diversity of DNA fingerprints inC. neoformans.J.Clin.Microbiol. 33, 1807–1814 (1995).

    PubMed  CAS  Google Scholar 

  • Vilgalys R., Hester M.: Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from severalCryptococcus species.J.Bacteriol. 172, 4238–4246 (1990).

    PubMed  CAS  Google Scholar 

  • Yamamoto Y., Shigeru K., Koga H., Kakeya H., Tomono K., Kaku M., Yamazaki T., Arisawa M., Hara K.: Random amplified polymorphic DNA analysis of clinically and environmentally isolatedC. neoformans in Nagasaki.J.Clin.Microbiol. 33, 3328–3332 (1995).

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Abdel-Salam, H.A. Characterization ofCryptococcus neoformans var.neoformans serotype A and A/D in samples from Egypt. Folia Microbiol 48, 261–268 (2003). https://doi.org/10.1007/BF02930967

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02930967

Keywords

Navigation