1887

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Volume 59, Issue 4

Molecular screening for and among Danish group blood cultureisolates: proposal of a new RFLP profile for differentiation Free

Abstract

and are recentlydescribed species phenotypically indistinguishable from . We evaluated phenotyping and molecular methods for the detection ofthese species among 79 unique blood culture isolates of the group obtained during the years 2004–2008. The isolates were screenedby PCR amplification of the secondary alcohol dehydrogenase-encoding gene () followed by digestion with the restriction enzyme I, using ATCC 22019, ATCC 96139 and ATCC 96144 as controls. Isolates withRFLP patterns distinct from were characterized bysequence analysis of the ITS1–ITS2, 26S rRNA (D1/D2) and regions. Restriction patterns for the 3 species with each of 610restriction enzymes were predicted using 12 available sequences.By PCR-RFLP of the gene alone, four isolates (5.1 %)had a pattern identical to the reference strain.Sequence analysis of and ITS (internal transcribed spacer)regions identified two of these isolates as . Theseresults were confirmed by creating a phylogenetic tree based on concatenatedsequences of , ITS and 26S rRNA gene sequence regions. Optimaldifferentiation between , and was predicted using digestion with III,producing discriminatory band sizes of: 131 and 505 bp; 74, 288 and 348 bp;and 131, 217 and 288 bp, respectively. This was confirmed using the referencestrains and 79 clinical isolates. In conclusion, reliable discrimination wasobtained by PCR-RFLP profile analysis of the gene after digestionwith III but not with I. and are involved in a small but significant numberof invasive infections in Denmark.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): AFLP, amplification fragment length polymorphism , AFST–EUCAST, Antifungal Susceptibility Testing Subcommittee–EuropeanCommittee on Antimicrobial Susceptibility Testing , ATCC, AmericanType Culture Collection , ITS, internal transcribed spacer and RAPD, randomly amplified polymorphic DNA
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2010-04-01
2023-06-09
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Molecular screening for Candida orthopsilosis and Candidametapsilosis among Danish Candida parapsilosis group blood cultureisolates: proposal of a new RFLP profile for differentiation
J. Med. Microbiol. 59, 414 (2010); https://doi.org/10.1099/jmm.0.017293-0
/content/journal/jmm/10.1099/jmm.0.017293-0
/content/journal/jmm/10.1099/jmm.0.017293-0
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