Experimental design for in-house validation of a screening immunoassay kit. The case of a multiplex dipstick for Fusarium mycotoxins in cereals

摘要: The benefits of using rapid qualitative methods to verify compliance food and feed with legislation requirements include user-friendly format, the possibility detection without expensive instrumentation, response affordable price. Prior their use, however, have pass validation experiments, in order assess performance profile. An experimental protocol for in-house a screening immunoassay has been designed applied evaluate characteristics multiplex dipstick kit determination major Fusarium toxins, namely zearalenone, T-2 HT-2 deoxynivalenol fumonisins wheat maize. test is intended cereals on presence/absence these mycotoxins at maximum permitted levels established by European or target levels. measurement determined reader device. Samples classified as negative are considered compliant, whereas positive samples need be re-analysed confirmatory methods. design consisted three steps, (1) estimating precision method including "between day" effects influences from different varieties matrices, (2) establishing robust cutoff values mycotoxin assuming an acceptable rate false results 5 % (3) assessment blank containing analytes below legal limits. total expressed relative standard deviation individually each analyte/concentration/matrix combination varied 9 30 was acceptable. In 17 out 28 cases, repeatability most important factor. predominance compared other factors (matrix, days) indicator ruggedness assay. study demonstrated that able differentiate contaminated lower than 6 %. Considering realistic occurrence samples, significant economical can expected when under real-world conditions.