title: Molecular tools for cryptic Candida species identification with applications in a clinical laboratory.
abstract: Candida spp. includes more than 160 species but only 20 species pose clinical problems. C. albicans and C. parapsilosis account for more than 75% of all the fungemias worldwide. In 1995 and 2005, one C. albicans and two C. parapsilosis-related species were described, respectively. Using phenotypic traits, the identification of these newly described species is inconclusive or impossible. Thus, molecular-based procedures are mandatory. In the proposed educational experiment we have adapted different basic molecular biology techniques designed to identify these species including PCR, multiplex PCR, PCR-based restriction endonuclease analysis and nuclear ribosomal RNA amplification. During the classes, students acquired the ability to search and align gene sequences, design primers, and use bioinformatics software. Also, in the performed experiments, fungal molecular taxonomy concepts were introduced and the obtained results demonstrated that classic identification (phenotypic) in some cases needs to be complemented with molecular-based techniques. As a conclusion we can state that we present an inexpensive and well accepted group of classes involving important concepts that can be recreated in any laboratory.
authors.affiliation: Laboratorio de Micología y Diagnóstico Molecular, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, CONICET, Santa Fe (CP 3000), Argentina.
authors.forename: Soledad
authors.initials: S
authors.lastname: Gamarra
authors.forename: Catiana
authors.initials: C
authors.lastname: Dudiuk
authors.forename: Estefanía
authors.initials: E
authors.lastname: Mancilla
authors.forename: María Verónica
authors.initials: MV
authors.lastname: Vera Garate
authors.forename: Sergio
authors.initials: S
authors.lastname: Guerrero
authors.forename: Guillermo
authors.initials: G
authors.lastname: Garcia-Effron
mesh_terms: D002175:Candida / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002176:Candida albicans / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D019295:Computational Biology / Q000193:education*; D017931:DNA Primers; D004262:DNA Restriction Enzymes / Q000737:chemistry; D006801:Humans; D007753:Laboratories; D008829:Microbiology / Q000193:education*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D016533:Mycological Typing Techniques / Q000379:methods*; D010802:Phylogeny; D012331:RNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D012335:RNA, Ribosomal / Q000235:genetics / Q000302:isolation & purification; D012984:Software; D013334:Students
chemical_list: D017931:DNA Primers; D012331:RNA, Fungal; D012335:RNA, Ribosomal; D004262:DNA Restriction Enzymes