COMPARISONMODIFICATIONS OF THE METHOD FOR DETECTION OF PLUM POX VIRUS DAS-ELISA [POROVNÁNÍ MODIFIKACÍ POSTUPU DETEKCE VIRU NEŠTOVIC PECKOVIN METODOU DAS-ELISA]

Abstract

During seasonal inspections and a need for detection of fruit tree pathogens, diagnostic laboratories often have to deal with situations where a large number of samples have to be analyzed in a short time. We therefore focused on the development of a faster and more efficient diagnostic procedure for the detection of Plum pox virus (PPV). A relatively cheap and simple serological method, the double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA), is usually used to detect PPV in plant material. The standard detection time for the DAS-ELISA kit sold by Bioreba AG is 32 hours. The optimization of the method involved reducing the incubation times and increasing the incubation temperature. At the same time, it was necessary to increase the antibody concentration to maintain the sensitivity of the method. Pathogen detection using a modified DAS-ELISA method from Bioreba AG can provide in one working day (8.5 h).

Keywords

• das-elisa
• diagnostics
• plum pox virus
• method optimization