Navegando por Autor "Ono, Elisabete Yurie Sataque"
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Item A comparison between enzyme immunoassay and HPLC for ochratoxin A detection in green, roasted and instant coffee(Instituto de Tecnologia do Paraná - Tecpar, 2007-03) Fujii, Simone; Ono, Elisabete Yurie Sataque; Ribeiro, Ricardo Marcelo Reche; Assunção, Fernanda Garcia Algarte; Takabayashi, Cássia Reika; Oliveira, Tereza Cristina Rocha Moreira de; Itano, Eiko Nakagawa; Ueno, Yoshio; Kawamura, Osamu; Hirooka, Elisa YokoAn indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for ochratoxin A (OTA) detection in green, roasted and instant coffees was developed using anti-OTA monoclonal antibody. Immunological reagents prepared were OTA-BSA (4.76 μ g/mL), anti-OTA.7 MAb (2x10 3 -fold dilution) and HRP-anti IgG (10 3 -fold dilution). The detection limit was 3.73 ng OTA/g and correlation coefficients (r) between this immunoassay and high performance liquid chromatography were 0.98 for green coffee, 0.98 for roasted and 0.86 for instant. OTA levels detected by ic- ELISA were higher than by HPLC, with ELISA/HPLC ratio of 0.66 - 1.46 (green coffee), 0.96 - 1.11 (roasted) and 0.93 - 1.82 (instant). ELISA recoveries for OTA added to coffee (5 - 70 ng/g) were 81.53 % for green coffee, 46.73 % for roasted and 64.35 % for instant, while recoveries by HPLC were 80.54 %, 45.91 % and 55.15 %, respectively. Matrices interferences were minimized by samples dilution before carrying out the ELISA assay. The results indicate that MAb-based ic-ELISA could be a simple, sensitive and specific screening tool for OTA detection, contributing to quality and safety of coffee products.Item Molecular identification of Aspergillus spp. isolated from coffee beans(Escola Superior de Agricultura "Luiz de Queiroz", 2005-01) Magnani, Marciane; Fernandes, Thiago; Prete, Cássio Egidio Cavenaghi; Homechim, Martin; Ono, Elisabete Yurie Sataque; Vilas-Boas, Laurival Antonio; Sartori, Daniele; Furlaneto, Márcia Cristina; Fungaro, Maria Helena PelegrinelliSome species belonging to the genus Aspergillus are potential producers of ochratoxin A (OA), a mycotoxin with nephrotoxic, immunosuppressive, teratogenic and carcinogenic effects. The aim of the present study was to identify the species of Aspergillus that contaminate the inside of coffee beans collected in the stage of maturation and drying, from 16 producing areas located in the northern region of the State of Paraná, in the South of Brazil. A total of 108 isolates of Aspergillus spp. was identified at the species level, by sequencing the internal transcribed spacer (ITS1-5.8S-ITS2) of ribosomal DNA (rDNA). The results revealed the presence of potentially ochratoxigenic species in 82% of the geographic regions studied, among which Aspergillus niger was the species most frequently detected, followed by A. ochraceus and A. carbonarius. The presence of A. carbonarius in immature coffee fruits harvested from trees is reported for the first time.