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Reliable indirect competitive ELISA used for a survey of ochratoxin A in green coffee from the North of Paraná State, Brazil.
Food Addit Contam. 2006 Sep; 23(9):902-9.FA

Abstract

The performance of an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a monoclonal antibody (mAb) for ochratoxin A (OTA) detection was evaluated in a comparative study with high-performance liquid chromatography (HPLC) analysis using 68 freshly harvested coffee samples from the North of Paraná State, Brazil. The anti-OTA mAb showed high specificity and low cross-reactivity with OTA analogues (OTB and OTalpha), but cross-reacted with OTC. This ic-ELISA showed a detection limit of 3.75 ngg-1 sample, when compared to 0.80 ngg-1 by HPLC, with an ic-ELISA/HPLC correlation coefficient of 0.90. As regards OTA analysis of these coffee samples, natural contamination was detected in 10 samples (14.7%) by both methods, where the ic-ELISA values (range 3.9-7.3 ngg-1) were 1.1 to 1.6-fold higher than HPLC data (2.7-4.7 ngg-1). Five samples (7.4%) were OTA positive (range 0.84-1.30 ngg-1) only by HPLC assay, probably due to the higher detection limit reached by ic-ELISA. OTA was undetectable in 53 samples (77.9%) by both methods, while all positive samples (range 0.84-7.30 ngg-1) showed OTA levels lower than 8 ngg-1 (maximum limit recommended by the European Union). The matrix interference of green coffee was minimized by dilution of sample extracts before carrying out the ELISA assay. This mAb-based ic-ELISA can be effectively applied for OTA screening in coffee, because it is simple, sensitive and sample preparation is easy.

Authors+Show Affiliations

Fujii S
Department of Food and Drug Technology, State University of Londrina, P.O.6001, 86051-990, Londrina, PR, Brazil.
Ribeiro RM
No affiliation info available
Scholz MB
No affiliation info available
Ono EY
No affiliation info available
Prete CE
No affiliation info available
Itano EN
No affiliation info available
Ueno Y
No affiliation info available
Kawamura O
No affiliation info available
Hirooka EY
No affiliation info available

MeSH

Antibodies, MonoclonalCarcinogensChromatography, High Pressure LiquidCoffeeEnzyme-Linked Immunosorbent AssayFood AnalysisFood ContaminationHumansMycotoxinsOchratoxinsReproducibility of Results

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16901859

Citation

Fujii, Simone, et al. "Reliable Indirect Competitive ELISA Used for a Survey of Ochratoxin a in Green Coffee From the North of Paraná State, Brazil." Food Additives and Contaminants, vol. 23, no. 9, 2006, pp. 902-9.
Fujii S, Ribeiro RM, Scholz MB, et al. Reliable indirect competitive ELISA used for a survey of ochratoxin A in green coffee from the North of Paraná State, Brazil. Food Addit Contam. 2006;23(9):902-9.
Fujii, S., Ribeiro, R. M., Scholz, M. B., Ono, E. Y., Prete, C. E., Itano, E. N., Ueno, Y., Kawamura, O., & Hirooka, E. Y. (2006). Reliable indirect competitive ELISA used for a survey of ochratoxin A in green coffee from the North of Paraná State, Brazil. Food Additives and Contaminants, 23(9), 902-9.
Fujii S, et al. Reliable Indirect Competitive ELISA Used for a Survey of Ochratoxin a in Green Coffee From the North of Paraná State, Brazil. Food Addit Contam. 2006;23(9):902-9. PubMed PMID: 16901859.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Reliable indirect competitive ELISA used for a survey of ochratoxin A in green coffee from the North of Paraná State, Brazil. AU - Fujii,Simone, AU - Ribeiro,Ricardo Marcelo Reche, AU - Scholz,Maria Brígida Dos Santos, AU - Ono,Elisabete Yurie Sataque, AU - Prete,Cássio Egidio C, AU - Itano,Eiko Nakagawa, AU - Ueno,Yoshio, AU - Kawamura,Osamu, AU - Hirooka,Elisa Yoko, PY - 2006/8/12/pubmed PY - 2007/3/16/medline PY - 2006/8/12/entrez SP - 902 EP - 9 JF - Food additives and contaminants JO - Food Addit Contam VL - 23 IS - 9 N2 - The performance of an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a monoclonal antibody (mAb) for ochratoxin A (OTA) detection was evaluated in a comparative study with high-performance liquid chromatography (HPLC) analysis using 68 freshly harvested coffee samples from the North of Paraná State, Brazil. The anti-OTA mAb showed high specificity and low cross-reactivity with OTA analogues (OTB and OTalpha), but cross-reacted with OTC. This ic-ELISA showed a detection limit of 3.75 ngg-1 sample, when compared to 0.80 ngg-1 by HPLC, with an ic-ELISA/HPLC correlation coefficient of 0.90. As regards OTA analysis of these coffee samples, natural contamination was detected in 10 samples (14.7%) by both methods, where the ic-ELISA values (range 3.9-7.3 ngg-1) were 1.1 to 1.6-fold higher than HPLC data (2.7-4.7 ngg-1). Five samples (7.4%) were OTA positive (range 0.84-1.30 ngg-1) only by HPLC assay, probably due to the higher detection limit reached by ic-ELISA. OTA was undetectable in 53 samples (77.9%) by both methods, while all positive samples (range 0.84-7.30 ngg-1) showed OTA levels lower than 8 ngg-1 (maximum limit recommended by the European Union). The matrix interference of green coffee was minimized by dilution of sample extracts before carrying out the ELISA assay. This mAb-based ic-ELISA can be effectively applied for OTA screening in coffee, because it is simple, sensitive and sample preparation is easy. SN - 0265-203X UR - https://www.unboundmedicine.com/medline/citation/16901859/Reliable_indirect_competitive_ELISA_used_for_a_survey_of_ochratoxin_A_in_green_coffee_from_the_North_of_Paraná_State_Brazil_ L2 - https://medlineplus.gov/foodborneillness.html DB - PRIME DP - Unbound Medicine ER -