Strain detection of Andean potato latent tymovirus
A new ELISA procedure has been described in Germany to detect the different strains of Andean potato latent tymovirus (EPPO A1 quarantine pest). The use of conventional DAS-ELISA has been limited in its use to detect the virus because of its strain specifity. Mixtures of antisera from different APLV strains used in a DAS-ELISA had overcome this strain specifity in detecting APLV. The newly developed ELISA procedure provided, according to the author, a higher sensitivity or a reduction of strain specifity and permits, therefore, the detection of APLV strains which more or less fail in DAS-ELISA. The biotin-streptavidin-alkaline phosphatase ELISA gave high extinction values also in heterologous combinations.
The biotin-streptavidin-alkaline phosphatase ELISA was carried out in the same manner as for DAS-ELISA, but instead of the AP-conjugate a biotin conjugate was used at a dilution of 1:1000 in conjugate buffer. The streptavidin-alkaline phosphatase conjugatewas diluted 1:10000 in conjugate buffer and incubated for 60 min at 37ø C. The enzym-substrate reaction was read after 30 minutes.
In comparison with the DAS-ELISA the biotin-ELISA gave higher absorption values from all strains and combinations of them.
Sources
Weidemann, H.L. (1993) Detection of different strains of Andean potato latent tymovirus by a biotin-streptavidin-ELISA.
Quartech, No. 1, 1993-07, 3-4.