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. 1995 Jun;33(6):1596–1601. doi: 10.1128/jcm.33.6.1596-1601.1995

Comparison of three typing methods for clinical and environmental isolates of Aspergillus fumigatus.

D Lin 1, P F Lehmann 1, B H Hamory 1, A A Padhye 1, E Durry 1, R W Pinner 1, B A Lasker 1
PMCID: PMC228223  PMID: 7650194

Abstract

To evaluate procedures used for epidemiologic analysis of outbreaks of aspergillosis, we analyzed a collection of 35 Aspergillus fumigatus isolates using three typing methods: isoenzyme analysis (IEA), random amplified polymorphic DNA (RAPD) analysis, and restriction endonuclease analysis (REA). Twenty-one isolates were from a single hospital, with four isolates coming from different patients. Three clinical isolates came from a different hospital, and 11 clinical or environmental isolates were derived from a culture collection. With IEA, the patterns of alkaline phosphatase, esterase, and catalase discriminated nine types. In contrast, 22 types were obtained with five different RAPD primers, and 21 types could be detected with three of these (R108, R151, and UBC90). Restriction endonuclease analysis of genomic DNA, digested with either XbaI, XhoI, or SalI, detected 3, 17, and 13 different REA types, respectively, and 22 types were identified by combining the data from the XhoI and SalI REAs. Twenty-eight types were obtainable with a combination of REA, IEA, and RAPD patterns. Overall, the results pointed to substantial genetic variation among the isolates. Though two isolates had markedly distinct genotypes, their morphologic features and exoantigens were consistent with their being A. fumigatus. The analysis will help in planning epidemiologic studies of aspergillosis.

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Selected References

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  1. Aufauvre-Brown A., Cohen J., Holden D. W. Use of randomly amplified polymorphic DNA markers to distinguish isolates of Aspergillus fumigatus. J Clin Microbiol. 1992 Nov;30(11):2991–2993. doi: 10.1128/jcm.30.11.2991-2993.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beyer J., Schwartz S., Heinemann V., Siegert W. Strategies in prevention of invasive pulmonary aspergillosis in immunosuppressed or neutropenic patients. Antimicrob Agents Chemother. 1994 May;38(5):911–917. doi: 10.1128/aac.38.5.911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Buffington J., Reporter R., Lasker B. A., McNeil M. M., Lanson J. M., Ross L. A., Mascola L., Jarvis W. R. Investigation of an epidemic of invasive aspergillosis: utility of molecular typing with the use of random amplified polymorphic DNA probes. Pediatr Infect Dis J. 1994 May;13(5):386–393. [PubMed] [Google Scholar]
  5. Burnie J. P., Coke A., Matthews R. C. Restriction endonuclease analysis of Aspergillus fumigatus DNA. J Clin Pathol. 1992 Apr;45(4):324–327. doi: 10.1136/jcp.45.4.324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burnie J. P., Matthews R. C., Clark I., Milne L. J. Immunoblot fingerprinting Aspergillus fumigatus. J Immunol Methods. 1989 Mar 31;118(2):179–186. doi: 10.1016/0022-1759(89)90004-5. [DOI] [PubMed] [Google Scholar]
  7. Denning D. W., Clemons K. V., Hanson L. H., Stevens D. A. Restriction endonuclease analysis of total cellular DNA of Aspergillus fumigatus isolates of geographically and epidemiologically diverse origin. J Infect Dis. 1990 Nov;162(5):1151–1158. doi: 10.1093/infdis/162.5.1151. [DOI] [PubMed] [Google Scholar]
  8. Denning D. W., Shankland G. S., Stevens D. A. DNA fingerprinting of Aspergillus fumigatus isolates from patients with aspergilloma. J Med Vet Mycol. 1991;29(5):339–342. [PubMed] [Google Scholar]
  9. 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]
  10. Fanti F., Conti S., Campani L., Morace G., Dettori G., Polonelli L. Studies on the epidemiology of Aspergillus fumigatus infections in a university hospital. Eur J Epidemiol. 1989 Mar;5(1):8–14. doi: 10.1007/BF00145038. [DOI] [PubMed] [Google Scholar]
  11. Girardin H., Sarfati J., Traoré F., Dupouy Camet J., Derouin F., Latgé J. P. Molecular epidemiology of nosocomial invasive aspergillosis. J Clin Microbiol. 1994 Mar;32(3):684–690. doi: 10.1128/jcm.32.3.684-690.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hadrys H., Balick M., Schierwater B. Applications of random amplified polymorphic DNA (RAPD) in molecular ecology. Mol Ecol. 1992 May;1(1):55–63. doi: 10.1111/j.1365-294x.1992.tb00155.x. [DOI] [PubMed] [Google Scholar]
  13. Hunter P. R. A critical review of typing methods for Candida albicans and their applications. Crit Rev Microbiol. 1991;17(6):417–434. doi: 10.3109/10408419109115206. [DOI] [PubMed] [Google Scholar]
  14. Hunter P. R., Gaston M. A. Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity. J Clin Microbiol. 1988 Nov;26(11):2465–2466. doi: 10.1128/jcm.26.11.2465-2466.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kemker B. J., Lehmann P. F., Lee J. W., Walsh T. J. Distinction of deep versus superficial clinical and nonclinical isolates of Trichosporon beigelii by isoenzymes and restriction fragment length polymorphisms of rDNA generated by polymerase chain reaction. J Clin Microbiol. 1991 Aug;29(8):1677–1683. doi: 10.1128/jcm.29.8.1677-1683.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kulik M. M., Brooks A. G. Electrophoretic studies of soluble proteins from Aspergillus spp. Mycologia. 1970 Mar-Apr;62(2):365–376. [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Leslie C. E., Flannigan B., Milne L. J. Morphological studies on clinical isolates of Aspergillus fumigatus. J Med Vet Mycol. 1988;26(6):335–341. [PubMed] [Google Scholar]
  20. Loudon K. W., Burnie J. P., Coke A. P., Matthews R. C. Application of polymerase chain reaction to fingerprinting Aspergillus fumigatus by random amplification of polymorphic DNA. J Clin Microbiol. 1993 May;31(5):1117–1121. doi: 10.1128/jcm.31.5.1117-1121.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Matsuda H., Kohno S., Maesaki S., Yamada H., Koga H., Tamura M., Kuraishi H., Sugiyama J. Application of ubiquinone systems and electrophoretic comparison of enzymes to identification of clinical isolates of Aspergillus fumigatus and several other species of Aspergillus. J Clin Microbiol. 1992 Aug;30(8):1999–2005. doi: 10.1128/jcm.30.8.1999-2005.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Penner G. A., Bush A., Wise R., Kim W., Domier L., Kasha K., Laroche A., Scoles G., Molnar S. J., Fedak G. Reproducibility of random amplified polymorphic DNA (RAPD) analysis among laboratories. PCR Methods Appl. 1993 May;2(4):341–345. doi: 10.1101/gr.2.4.341. [DOI] [PubMed] [Google Scholar]
  23. Sekhon A. S., Standard P. G., Kaufman L., Garg A. K., Cifuentes P. Grouping of Aspergillus species with exoantigens. Diagn Immunol. 1986;4(2):112–116. [PubMed] [Google Scholar]
  24. Spreadbury C. L., Bainbridge B. W., Cohen J. Restriction fragment length polymorphisms in isolates of Aspergillus fumigatus probed with part of the intergenic spacer region from the ribosomal RNA gene complex of Aspergillus nidulans. J Gen Microbiol. 1990 Oct;136(10):1991–1994. doi: 10.1099/00221287-136-10-1991. [DOI] [PubMed] [Google Scholar]
  25. Tang C. M., Cohen J., Rees A. J., Holden D. W. Molecular epidemiological study of invasive pulmonary aspergillosis in a renal transplantation unit. Eur J Clin Microbiol Infect Dis. 1994 Apr;13(4):318–321. doi: 10.1007/BF01974610. [DOI] [PubMed] [Google Scholar]
  26. Walsh T. J., Dixon D. M. Nosocomial aspergillosis: environmental microbiology, hospital epidemiology, diagnosis and treatment. Eur J Epidemiol. 1989 Jun;5(2):131–142. doi: 10.1007/BF00156818. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Weems J. J., Jr, Andremont A., Davis B. J., Tancrede C. H., Guiguet M., Padhye A. A., Squinazi F., Martone W. J. Pseudoepidemic of aspergillosis after development of pulmonary infiltrates in a group of bone marrow transplant patients. J Clin Microbiol. 1987 Aug;25(8):1459–1462. doi: 10.1128/jcm.25.8.1459-1462.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Welsh J., McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 1990 Dec 25;18(24):7213–7218. doi: 10.1093/nar/18.24.7213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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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