Evaluation of Data Competency Achieved from Double Sandwich ELISA Method for the Detection of Botulinum Toxin Type

Botulinum toxin is known as the most potent toxin that that can cause death even in small amounts، on the other hand، except antitoxin، there are no drug for toxicity. Therefore، diagnostic methods that can detect very small amounts of botulinum toxin in a short time، is very important. The aim of this study was to evaluate the modified double sandwich ELISA method for detection limit، repeatability and cross-reactivity for the detection of botulinum toxin type E. Double-sandwich ELISA method require two antibodies that bind to epitopes without interference on antigens. For this purpose، we poured the rabbit antibody at down (Capture antibody) and mouse antibody at high toxin (Detection antibody) into the each wells and titered toxin from the first well. Then، for adjusting and optimizing the sandwich ELISA system Cross reactivity and Reproducibility tests were performed. Double-sandwich ELISA method is able to detect value of 1 ngbotulinum toxin type E. Reproducibility of this method was optimum through experiments repeated three times in one day and four times in different days and is negligible cross it''s reactivity with toxin type B and Tetanus was insignificant. So far، several methods based on serological assessment of botulinum toxin have been presented، but ELISA، has been the best and most practical method. To improve sensitivity of assessment، several modified ELISA methods have been presented that double-sandwich ELISA is the most advanced one. This method is rapid، highly specific and it''s sensitivity is equivalent to testing on mice. The method is used for determining specific antigen in the unknown sample.