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Journal of Clinical Microbiology, February 2001, p. 658-669, Vol. 39, No. 2
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.2.658-669.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Cloning and Characterization of a Complex DNA
Fingerprinting Probe for Candida parapsilosis
Lee
Enger,1
Sophie
Joly,1
Claude
Pujol,1
Patricia
Simonson,1
Michael
Pfaller,2 and
David R.
Soll1,*
Department of Biological
Sciences1 and
Pathology,2 University of Iowa, Iowa
City, Iowa 52242
Received 5 September 2000/Returned for modification 13 November
2000/Accepted 29 November 2000
Candida parapsilosis accounts for a significant number
of nosocomial fungemias, but in fact, no effective and verified genetic fingerprinting method has emerged for assessing the relatedness of
independent isolates for epidemiological studies. A complex 15-kb DNA
fingerprinting probe, Cp3-13, was therefore isolated from a library of
C. parapsilosis genomic DNA fragments. The efficacy of
Cp3-13 for DNA fingerprinting was verified by a comparison of its
clustering capacity with those of randomly amplified polymorphic DNA
analysis and internally transcribed spacer region sequencing, by
testing species specificity, and by assessing its capacity to identify
microevolutionary changes both in vitro and in vivo. Southern blot
hybridization of EcoRI/SalI-digested DNA with
Cp3-13 provides a fingerprinting system that (i) identifies the same strain in independent isolates, (ii) discriminates between unrelated isolates, (iii) separates independent isolates into valid groups in a
dendrogram, (iv) identifies microevolution in infecting populations, and (v) is amenable to automatic computer-assisted DNA fingerprint analysis. This probe is now available for epidemiological studies.
*
Corresponding author. Mailing address: Room 302 BBE,
Department of Biological Sciences, University of Iowa, Iowa City, IA 52242. Phone: (319) 335-1117. Fax: (319) 335-2772. E-mail:
david-soll{at}uiowa.edu.
Journal of Clinical Microbiology, February 2001, p. 658-669, Vol. 39, No. 2
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.2.658-669.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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