Systematic internal transcribed spacer sequence analysis for identification of clinical mold isolates in diagnostic mycology: a 5-year study.
The implementation of internal transcribed spacer (ITS) sequencing for routine identification of molds in the diagnostic mycology laboratory was analyzed in a 5-year study. All mold isolates (n = 6,900) recovered in our laboratory from 2005 to 2009 were included in this study. According to a defined work flow, which in addition to troublesome phenotypic identification takes clinical relevance into account, 233 isolates were subjected to ITS sequence analysis. Sequencing resulted in successful identification for 78.6% of the analyzed isolates (57.1% at species level, 21.5% at genus level). In comparison, extended in-depth phenotypic characterization of the isolates subjected to sequencing achieved taxonomic assignment for 47.6% of these, with a mere 13.3% at species level. Optimization of DNA extraction further improved the efficacy of molecular identification. This study is the first of its kind to testify to the systematic implementation of sequence-based identification procedures in the routine workup of mold isolates in the diagnostic mycology laboratory.
Ciardo
Diana E
DE
Institut für Medizinische Mikrobiologie, Universität Zürich, Gloriastr. 30/32, 8006 Zürich, Switzerland.
Lucke
Katja
K
Imhof
Alex
A
Bloemberg
Guido V
GV
Böttger
Erik C
EC
D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics*; D005658:Fungi / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016533:Mycological Typing Techniques / Q000379:methods; D009172:Mycology / Q000379:methods*; D009181:Mycoses / Q000175:diagnosis* / Q000382:microbiology; D012680:Sensitivity and Specificity; D017421:Sequence Analysis / Q000379:methods*
D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer