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Single primer pair for PCR identification of Candida parapsilosis group I isolates

EUGENIO PONTIERI, CLAUDIA CARACCIOLO, STEFANO BIANCHINI, DOMENICO D'ANTONIO^*, GIUSEPPE NOVELLI, BRUNO DALLAPICCOLA and GIUSEPPE CARRUBA

Cattedra di Virologia e Micologia Medica, Dipartimento di Medicina Sperimentale, Università degli Studi, l'Aquila, *Servizio di Microbiologia Clinica, Dipartimento di Ematologia ed Oncologia, Ospedale ‘Santo Spirito', Pescara, Cattedra di Genetica Umana Dipartimento di Biopatologia e Diagnostica per Immagini, Università di Roma ‘Tor Vergata', Cattedra di Genetica Umana Università La Sapienza Roma and Istituto CSS-Mendel, Rome, Italy

Corresponding author: Professor G. Carruba (e-mail: carruba{at}fismedw2.univaq.it).

Received 2 May 2000; revised version accepted 10 Nov. 2000.

Abstract

A DNA fragment, isolated from a genomic DNA mini-library of Candida parapsilosis group I reference strain ATCC 22019, was sequenced and characterised. The fragment was first probed by Southern blotting against a pool of DNA from several yeasts. The hybridisation tests revealed that the probe was specific for strain ATCC 22019 and 49 (90.74%) of 54 C. parapsilosis clinical and soil isolates that were attributed to C. parapsilosis group I by the electrophoretic images of their restriction fragment length polymorphisms (RFLPs) and electrophoretic karyotype (EK). The remaining five negative isolates, according to the same criteria, were attributed to group II (one isolate) and III (four isolates). Two primers were selected from the probe DNA sequence and a PCR-based test was developed for specifically detecting C. parapsilosis group I isolates, which represent the majority of the common clinical isolates. The PCR assay confirmed the Southern hybridisation results. This PCR assay could be a simple and reproducible tool for the rapid, sensitive and species-specific identification of C. parapsilosis major group I isolates.

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