Adelaida Gaviria-Rivera1, Alejandra Giraldo-López2, Carolina Santa-Cardona3, Luz Cano-Restrepo4. 1. AG: Ing. Agrónoma. Ph. D. Biological Sciences. Escuela de Biociencias. Facultad de Ciencias. Universidad Nacional de Colombia. Medellín, Colombia. amgavirr@unal.edu.co. 2. AG: Bacterióloga. M. Sc. Biotecnología. Ph. D. Ciencias Médicas Básicas. Westerdijk Fungal Biodiversity Institute. Utrecht, the Netherlands. a.giraldo@westerdijkinstitute.nl. 3. CS: Biotecnóloga. Colegio Mayor de Antioquia. Medellín, Colombia. santa8612@gmail.com. 4. LC: Téc. Laboratorio Clínico. Lic. Bacteriología y Laboratorio Clínico. Ph. D. Ciencias. Escuela de Bacteriología y Laboratorio Clínico, Universidad de Antioquia. Medellín. Grupo de Micología Médica y Experimental, CIB-UDEA-UPB. Corporación para Investigaciones Biológicas. Escuela de Microbiología, UdeA. Escuela de la Salud, UPB. Medellín, Colombia. lcano@cib.org.co; luz.Cano@udea.edu.co.
Abstract
OBJECTIVE: Identifying Fusarium isolates from mycosis symptomatic patients through molecular techniques as PCR and sequencing. METHODS: In this study, samples were taken from 101 mycosis symptomatic patients in-between 2004-2006. To determine isolates belonging to the Fusarium genus, the DNAr 28S region was amplified through PCR and specific PCR primers further confirmed their identity to the species level. Additionally, in order to confirm the identity of the species of the isolates, 75 isolates of these were analyzed by partial sequencing of the 28S rDNA and the TEF1-α gene. RESULTS: The 28S rDNA portion detected all 101 isolates as belonging to Fusarium and the PCR specific primers detected 52 and 29 isolates as F. oxysporum and F. solani, respectively; 34 and 41 of these, afterwards studied by partial sequencing of the 28S rDNA and TEF1- α genes respectively, were effectively identified by the technique. CONCLUSION: From all the molecular markers used to identify Fusarium isolates, the sequence of the TEF1-α gene provided the best resolution in the identification of species level; however it is possible to discriminate between F. oxysporum and F. solani isolates by PCR, in most of the cases, what is important considering the simplicity of the technique and a faster diagnosis.
OBJECTIVE: Identifying Fusarium isolates from mycosis symptomatic patients through molecular techniques as PCR and sequencing. METHODS: In this study, samples were taken from 101 mycosis symptomatic patients in-between 2004-2006. To determine isolates belonging to the Fusarium genus, the DNAr 28S region was amplified through PCR and specific PCR primers further confirmed their identity to the species level. Additionally, in order to confirm the identity of the species of the isolates, 75 isolates of these were analyzed by partial sequencing of the 28S rDNA and the TEF1-α gene. RESULTS: The 28S rDNA portion detected all 101 isolates as belonging to Fusarium and the PCR specific primers detected 52 and 29 isolates as F. oxysporum and F. solani, respectively; 34 and 41 of these, afterwards studied by partial sequencing of the 28S rDNA and TEF1- α genes respectively, were effectively identified by the technique. CONCLUSION: From all the molecular markers used to identify Fusarium isolates, the sequence of the TEF1-α gene provided the best resolution in the identification of species level; however it is possible to discriminate between F. oxysporum and F. solani isolates by PCR, in most of the cases, what is important considering the simplicity of the technique and a faster diagnosis.