Immunoassay of ochratoxin and other mycotoxins from a single extract of cereal grains utilizing monoclonal antibodies.
Immunoassays provide rapid, specific, sensitive and inexpensive methods for analysing mycotoxins but have generally been tested individually. Mycotoxigenic fungi rarely occur in pure culture in nature, and different mycotoxins may occur together. It would therefore be advantageous if several immunoassays for different mycotoxins could utilize a single extract. Monoclonal antibodies specific for ochratoxin A, aflatoxin B1 and T-2 toxin, raised at the University of Strathclyde, allow detection limits of 1 ng/ml ochratoxin A, 0.1 ng/ml aflatoxin B1 and 10 ng/ml T-2 toxin, when used in competitive enzyme-linked immunosorbent assays. These antibodies have been used to assay ochratoxin A. aflatoxin B1 and T-2 toxin in a single acetonitrile: 0.5% KCl:6% H2SO4 (89:10:1) extract of cereal grain. Extracts were either diluted 1:10 for direct assay or subjected, before assay, to a simple liquid-liquid clean-up procedure, which removed interfering substances and resulted in a 5:1 concentration. Recoveries from barley to which mycotoxins had been added averaged 95.8% for ochratoxin A, 93.8% for aflatoxin B1 and 80.6% for T-2 toxin, and the detection limits were 5 ng/g for ochratoxin A, 4 ng/g for aflatoxin B1 and 50 ng/g for T-2 toxin. Mean coefficients of variation within and between assays and between subsamples were less than 12% for ochratoxin A and aflatoxin B1 but up to 17% for T-2 toxin in assays of barley inoculated with toxigenic fungi.(ABSTRACT TRUNCATED AT 250 WORDS)
Lacey
J
J
AFRC Institute of Arable Crops Research, Rothamsted Experimental Station, Harpenden, Herts, United Kingdom.
Ramakrishna
N
N
Candlish
A A
AA
Smith
J E
JE
D000097:Acetonitriles; D016604:Aflatoxin B1 / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001231:Aspergillus flavus / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005504:Food Analysis / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D005670:Fusarium / Q000378:metabolism; D001467:Hordeum / Q000737:chemistry* / Q000382:microbiology; D009793:Ochratoxins / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D010407:Penicillium / Q000378:metabolism; D012997:Solvents; D013605:T-2 Toxin / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification
D000097:Acetonitriles; D000911:Antibodies, Monoclonal; D009793:Ochratoxins; D012997:Solvents; C025589:ochratoxin A; D016604:Aflatoxin B1; D013605:T-2 Toxin; C032159:acetonitrile