Efficient methods for conjugation of mycotoxins to proteins have been developed in the early phase of this study. Specific antibodies against different mycotoxins including aflatoxins B?1?, B?2a?, B1-diol, G?1?, M?1?, Q?1?, ochratoxin A, sterigmatocystin, and trichothecene mycotoxins such as T-2 toxin, diacetoxyscirpenol, deoxynivalenol and deoxyverrucarol have been obtained in our laboratory in the past few years. Antibodies obtained from rabbits after immunizing conjugates that were prepared by linking different side chains in the aflatoxin and trichothecene molecules to protein showed a diversified specificity. Antibodies against aflatoxins B?2a? and B1-diol were also found to be reactive with aflatoxin-DNA adduct. With the availability of these antibodies, simple and rapid, i.e., no cleanup treatment of samples to be assayed, radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) for the determination of these mycotoxins in grains and biological fluids have been developed. The sensitivity of RIA was generally in the range of 0.25 to 0.5 ng per assay. ELISA was found to be more sensitive than RIA, with a detection limit in the range of 2.5 to 25 pg per assay. Pretreatment of milk and human urine samples with a Sep-Pak, C-18 reversed-phase cartridge permits detection by direct ELISA of 10 to 25 ppt of aflatoxin M?1? in the sample. This technique was found to be very useful for monitoring aflatoxin M?1? in human urine samples obtained from the subjects who had consumed aflatoxin B?1?. A direct correlation between the dietary level of aflatoxin B?1? and aflatoxin M?1? excreted in human urine in a selected high incidence liver cancer region has been observed. In addition, immunohistochemical techniques for monitoring the aflatoxin, ochratoxin, and T-2 toxin in the tissues of mice and rats that have ingested these mycotoxins have been developed. (HI)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA015064-14
Application #
3164081
Study Section
Bacteriology and Mycology Subcommittee 1 (BM)
Project Start
1979-09-01
Project End
1990-12-31
Budget Start
1986-01-01
Budget End
1986-12-31
Support Year
14
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Wisconsin Madison
Department
Type
Earth Sciences/Resources
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Park, J J; Chu, F S (1996) Assessment of immunochemical methods for the analysis of trichothecene mycotoxins in naturally occurring moldy corn. J AOAC Int 79:465-71
Park, J J; Chu, F S (1996) Partial purification and characterization of an esterase from Fusarium sporotrichioides. Nat Toxins 4:108-16
Park, J J; Smalley, E B; Chu, F S (1996) Natural occurrence of Fusarium mycotoxins in field samples from the 1992 Wisconsin corn crop. Appl Environ Microbiol 62:1642-8
Chu, F S (1992) Recent progress on analytical techniques for mycotoxins in feedstuffs. J Anim Sci 70:3950-63
Yu, J H; Chu, F S (1991) Immunochromatography of fusarochromanone mycotoxins. J Assoc Off Anal Chem 74:655-60
Chu, F S (1991) Mycotoxins: food contamination, mechanism, carcinogenic potential and preventive measures. Mutat Res 259:291-306
Chu, F S (1990) Immunoassays for mycotoxins: current state of the art, commercial and epidemiological applications. Vet Hum Toxicol 32 Suppl:42-50
Lee, R C; Wei, R D; Chu, F S (1989) Enzyme-linked immunosorbent assay for T-2 toxin metabolites in urine. J Assoc Off Anal Chem 72:345-8
Zhang, G; Chu, F S (1989) Production and characterization of antibodies cross-reactive with major aflatoxins. Experientia 45:182-4
Chu, F S; Lee, R C; Trucksess, M W et al. (1988) Evaluation of enzyme-linked immunosorbent assay of cleanup for thin-layer chromatography of aflatoxin B1 in corn, peanuts, and peanut butter. J Assoc Off Anal Chem 71:953-6

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