1887

Abstract

Summary

A 317-base pair (bp) fragment of the heat shock protein 90 (HSP 90) gene was amplified by the polymerase chain reaction (PCR) for detection of DNA in clinical specimens. One hundred specimens were examined including swabs (39), urines (36), peritoneal fluid (9), pus (8) and blood or serum (8): 23% gave positive results with routine culture, 31% with extended broth culture and 37% with PCR. The amplified product was identified by hybridisation with a radiolabelled internal probe and their restriction enzyme digest patterns (I, III, RI, I and I), which could be predicted from the known sequence of HSP 90. DNA gave the characteristic 317-bp band and specifically hybridised with restriction enzyme-digested candidal DNA. DNA from other sources intermittently gave multiple faint bands especially in the presence of high concentrations of DNA, but these could be readily distinguished. The method was sensitive to 50 pg of DNA (5 pg with radiolabelled probing) and 100 cfu of .

Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-39-3-233
1993-09-01
2024-04-23
Loading full text...

Full text loading...

/deliver/fulltext/jmm/39/3/medmicro-39-3-233.html?itemId=/content/journal/jmm/10.1099/00222615-39-3-233&mimeType=html&fmt=ahah

References

  1. Edwards JE. Invasive Candida infections. Evolution of a fungal pathogen. N Engl J Med 1991; 324:1060–1062
    [Google Scholar]
  2. de Repentigny L, Reiss E. Current trends in immunodiagnosis of candidiasis and aspergillosis. Rev Infect Dis 1984; 6:301–312
    [Google Scholar]
  3. Wey SB, Mori M, Pfaller M, Woolson RF, Wenzel RP. Hospital-acquired candidemia: the attributable mortality and excess length of stay. Arch Intern Med 1988; 148:2642–2645
    [Google Scholar]
  4. Koenig H, Walle RJ, Krener M. Diagnosis and epidemiology of 70,000 isolated yeasts. Rev Fr Lab 1989; 197:34–40
    [Google Scholar]
  5. Graybill JR. Future directions of antifungal chemotherapy. Clin Infect Dis 1992; 14:Suppl 1S170–181
    [Google Scholar]
  6. Gold JWM, Wong B, Bernard EM, Kiehn TE, Armstrong D. Serum arabinitol concentrations and arabinitol/creatinine ratios in invasive candidiasis. J Infect Dis 1983; 147:504–513
    [Google Scholar]
  7. Walsh TJ, Hathorn JW, Sobel JD et al. Detection of circulating Candida enolase by immunoassay in patients with cancer and invasive candidiasis. N Engl J Med 1991; 324:1026–1031
    [Google Scholar]
  8. Matthews RC, Burnie JP. Diagnosis of systemic candidiasis by an enzyme-linked dot immunobinding assay for a circulating immunodominant 47-kilodalton antigen. J Clin Microbiol 1988; 26:459–463
    [Google Scholar]
  9. Buchman TG, Rossier M, Merz WG, Charache P. Detection of surgical pathogens by in vitro DNA amplification. Part 1. Rapid identification of Candida albicans by in vitro amplification of a fungus-specific gene. Surgery 1990; 108:338–345
    [Google Scholar]
  10. Matthews RC, Bumie JP. Cloning of a DNA sequence encoding a major fragment of the 47-kilodalton stress protein hom-ologue of Candida albicans. FEMS Microbiol Lett 1989; 60:25–30
    [Google Scholar]
  11. Rebbe NF, Ware J, Bertina RM, Modrich P, Stafford DW. Nucleotide sequence of a cDNA for a member of the human 90-kDa heat-shock protein family. Gene 1987; 53:235–245
    [Google Scholar]
  12. Borkovitch KA, Farrelly FW, Finkelstein DB, Taulien J, Lindquist S. HSP82 is an essential protein that is required in higher concentrations for growth of cells at higher temperatures. Mol Cell Biol 1989; 9:3919–3930
    [Google Scholar]
  13. Burnie JP, Odds FC, Lee W, Webster C, Williams JD. Outbreak of systemic Candida albicans in intensive care unit caused by cross infection. BMJ 1985; 290:746–748
    [Google Scholar]
  14. Maniatis T, Sambrook J, Fritsch EF. (eds) Molecular cloning: A laboratory manual. New York City: Cold Spring Harbour Laboratory Press;
    [Google Scholar]
  15. Vogelstein B, Gillespie D. Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci USA 1979; 76:615–619
    [Google Scholar]
  16. Innis MA, Gelfand DH. Optimization of PCRs. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ. (eds) PCR Protocols: A guide to methods and applications San Diego: Academic Press, Inc.; 19903–12
    [Google Scholar]
  17. Taylor GR. Polymerase chain reaction: basic principles and automation. In: McPherson MJ, Quirke P, Taylor GR. (eds) PCR—a practical approach Oxford: IRL Press; 19921–14
    [Google Scholar]
  18. Sarkar G, Sommer SS. Shedding light on PCR contamination. Nature 1990; 343:27
    [Google Scholar]
  19. Burnie JP, Coke A, Matthews RC. Restriction endonuclease analysis of Aspergillus fumigatus DNA. J Clin Pathol 1992; 45:324–327
    [Google Scholar]
  20. Skjold SA, Quie PG, Fries LA, Barnham M, Clery PP. DNA fingerprinting of Streptococcus zooepidermicus (Lancefield group C) as an aid to epidemiological study. J Infect Dis 1987; 155:1145–1150
    [Google Scholar]
  21. Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1984; 137:266–267
    [Google Scholar]
  22. 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; 30:894–900
    [Google Scholar]
  23. Holmes AR, Lee YC, Cannon RD, Jenkinson HF, Shepherd MG. Yeast-specific DNA probes and their application for the detection of Candida albicans. J Med Microbiol 1992; 37:346–351
    [Google Scholar]
  24. Kan V, Bennett JE. Polymerase chain reaction for deep candidiasis. 32nd ICAAC Anaheim, USA 1992 Abstract no: 1627
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-39-3-233
Loading
/content/journal/jmm/10.1099/00222615-39-3-233
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error