Rapid PCR-based test for identifying Candida albicans by using primers derived from the pH-regulated KER1 gene.
A PCR-based method in combination with a simple, reliable and inexpensive DNA extraction procedure for rapid detection of Candida albicans clinical isolates is described here. The extraction protocol is based on a combination of chemical (NaOH and detergents) and physical (boiling) treatments, thus avoiding many of the problems inherent in the currently available DNA extraction protocols (basically the use of expensive and/or toxic chemical reagents), and may be useful for daily clinical routine. The PCR-based system described here uses a single pair of primers (SC1F and SC1R) deduced from the C. albicans-specific KER1 gene sequence. These primers amplify a 670-bp fragment of the KER1 gene. All the clinical C. albicans isolates generated the expected 670-bp amplicon. Other non-albicans Candida species, including the azole-resistant C. krusei and C. glabrata, and the very closely related C. dubliniensis, failed to amplify any DNA fragment. The PCR results reported here suggest that amplification with SC1F and SC1R primers is species specific and, consequently, may be useful for specifically identifying C. albicans strains.
Galán
Amparo
A
Banco de líneas celulares, Centro de Investigación Príncipe Felipe, Valencia, Spain.
Veses
Verónica
V
Murgui
Amelia
A
Casanova
Manuel
M
Martínez
José P
JP
D002176:Candida albicans / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D018698:Glutamic Acid / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000737:chemistry; D008565:Membrane Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D005656:Fungal Proteins; D008565:Membrane Proteins; D018698:Glutamic Acid; D008239:Lysine