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. 1995 Mar;33(3):625–628. doi: 10.1128/jcm.33.3.625-628.1995

Improved detection of Candida albicans by PCR in blood of neutropenic mice with systemic candidiasis.

A J van Deventer 1, W H Goessens 1, A van Belkum 1, H J van Vliet 1, E W van Etten 1, H A Verbrugh 1
PMCID: PMC228002  PMID: 7751367

Abstract

A PCR using primers aimed at the multicopy gene coding for the small subunit rRNA and resulting in the synthesis of a 180-bp fragment was evaluated for its use in diagnosing invasive candidiasis in comparison with blood culture. With the use of a C. albicans-specific probe, +/- 10 to 15 C. albicans cells are detected in 100 microliters of whole blood by Southern analysis. A DNase pretreatment was critical in the purification process of yeast DNA from whole blood. Omission of the DNase pretreatment decreased assay sensitivity 10-fold. PCR analysis of blood specimens collected from mice with invasive candidiasis is more sensitive than blood culture (100 versus 67%, respectively) at 72 h after intravenous (i.v.) inoculation with C. albicans. Furthermore, the intensity of the hybridization signals increased with the progression of infection. In contrast, multiple blood samples from gastrointestinally colonized mice were all negative by PCR, indicating that the PCR assay is also specific and may, therefore, make a positive contribution to the detection and follow-up of invasive candidiasis.

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Selected References

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  1. Anaissie E. Opportunistic mycoses in the immunocompromised host: experience at a cancer center and review. Clin Infect Dis. 1992 Mar;14 (Suppl 1):S43–S53. doi: 10.1093/clinids/14.supplement_1.s43. [DOI] [PubMed] [Google Scholar]
  2. Berenguer J., Buck M., Witebsky F., Stock F., Pizzo P. A., Walsh T. J. Lysis-centrifugation blood cultures in the detection of tissue-proven invasive candidiasis. Disseminated versus single-organ infection. Diagn Microbiol Infect Dis. 1993 Aug-Sep;17(2):103–109. doi: 10.1016/0732-8893(93)90020-8. [DOI] [PubMed] [Google Scholar]
  3. Buchman T. G., Rossier M., Merz W. G., Charache P. Detection of surgical pathogens by in vitro DNA amplification. Part I. Rapid identification of Candida albicans by in vitro amplification of a fungus-specific gene. Surgery. 1990 Aug;108(2):338–347. [PubMed] [Google Scholar]
  4. Crampin A. C., Matthews R. C. Application of the polymerase chain reaction to the diagnosis of candidosis by amplification of an HSP 90 gene fragment. J Med Microbiol. 1993 Sep;39(3):233–238. doi: 10.1099/00222615-39-3-233. [DOI] [PubMed] [Google Scholar]
  5. Holmes A. R., Cannon R. D., Shepherd M. G., Jenkinson H. F. Detection of Candida albicans and other yeasts in blood by PCR. J Clin Microbiol. 1994 Jan;32(1):228–231. doi: 10.1128/jcm.32.1.228-231.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hopfer R. L., Walden P., Setterquist S., Highsmith W. E. Detection and differentiation of fungi in clinical specimens using polymerase chain reaction (PCR) amplification and restriction enzyme analysis. J Med Vet Mycol. 1993;31(1):65–75. doi: 10.1080/02681219380000071. [DOI] [PubMed] [Google Scholar]
  7. Jones J. M. Laboratory diagnosis of invasive candidiasis. Clin Microbiol Rev. 1990 Jan;3(1):32–45. doi: 10.1128/cmr.3.1.32. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kan V. L. Polymerase chain reaction for the diagnosis of candidemia. J Infect Dis. 1993 Sep;168(3):779–783. doi: 10.1093/infdis/168.3.779. [DOI] [PubMed] [Google Scholar]
  9. Makimura K., Murayama S. Y., Yamaguchi H. Detection of a wide range of medically important fungi by the polymerase chain reaction. J Med Microbiol. 1994 May;40(5):358–364. doi: 10.1099/00222615-40-5-358. [DOI] [PubMed] [Google Scholar]
  10. Miyakawa Y., Mabuchi T., Fukazawa Y. New method for detection of Candida albicans in human blood by polymerase chain reaction. J Clin Microbiol. 1993 Dec;31(12):3344–3347. doi: 10.1128/jcm.31.12.3344-3347.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Miyakawa Y., Mabuchi T., Kagaya K., Fukazawa Y. Isolation and characterization of a species-specific DNA fragment for detection of Candida albicans by polymerase chain reaction. J Clin Microbiol. 1992 Apr;30(4):894–900. doi: 10.1128/jcm.30.4.894-900.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Murray P. R. Comparison of the lysis-centrifugation and agitated biphasic blood culture systems for detection of fungemia. J Clin Microbiol. 1991 Jan;29(1):96–98. doi: 10.1128/jcm.29.1.96-98.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Niesters H. G., Goessens W. H., Meis J. F., Quint W. G. Rapid, polymerase chain reaction-based identification assays for Candida species. J Clin Microbiol. 1993 Apr;31(4):904–910. doi: 10.1128/jcm.31.4.904-910.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. van Etten E. W., van de Rhee N. E., van Kampen K. M., Bakker-Woudenberg I. A. Effects of amphotericin B and fluconazole on the extracellular and intracellular growth of Candida albicans. Antimicrob Agents Chemother. 1991 Nov;35(11):2275–2281. doi: 10.1128/aac.35.11.2275. [DOI] [PMC free article] [PubMed] [Google Scholar]

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