An interlaboratory comparison of ITS2-PCR for the identification of yeasts, using the ABI Prism 310 and CEQ8000 capillary electrophoresis systems.
BACKGROUND
Currently, most laboratories identify yeasts routinely on the basis of morphology and biochemical reactivity. This approach has quite often limited discriminatory power and may require long incubation periods. Due to the increase of fungal infections and due to specific antifungal resistence patterns for different species, accurate and rapid identification has become more important. Several molecular techniques have been described for fast and reliable identification of yeast isolates, but interlaboratory exchangeability of identification schemes of molecular techniques has hardly been studied. Here, we compared amplified ITS2 fragment length determination by an ABI Prism 310 (Applied Biosystems, Foster City, Ca.) capillary electrophoresis system with that obtained by a CEQ8000 (Beckman Coulter, Fullerton, Ca.) capillary electrophoresis system.


RESULTS
Although ITS2 size estimations on both systems differed and separate libraries had to be constructed for each system, both approaches had the same discriminatory power with regard to the 44 reference strains, identical identifications were obtained for 39/ 40 clinical isolates in both laboratories and strains from 51 samples were correctly identified using CEQ8000, when compared to phenotypic identification.


CONCLUSIONS
Identification of yeasts with ITS2-PCR followed by fragment analysis can be carried out on different capillary electrophoresis systems with comparable discriminatory power.
De Baere
Thierry
T
Department of Clinical Chemistry, Microbiology and Immunology, University Hospital Ghent, Belgium. Thierry.Debaere@Ugent.be
Van Keerberghen
Anne
A
Van Hauwe
Peter
P
De Beenhouwer
Hans
H
Boel
An
A
Verschraegen
Gerda
G
Claeys
Geert
G
Vaneechoutte
Mario
M
D004271:DNA, Fungal / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods*; D015003:Yeasts / Q000235:genetics* / Q000302:isolation & purification*
D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer