Type of detection:
Serological
Type of method:
DAS-ELISA
Description of the test:
DAS-ELISA test to detect PepMV (6). PepMV, a potexvirus, was first reported in pepino (Solanum muricatum) in Peru (2). In Europe, first outbreaks of PepMV in greenhouse tomato have been reported 1999/2000 in the Netherlands, Germany, France, the UK and Spain, subsequently in many European countries as well as in Canada, USA and China. Other solanaceous plants such as potatoes could be infected by artificial inoculation, but so far no PepMV problem has been observed in that crop. Different PepMV genotypes have been recognized (3, 5). Transmission occurs by contact, e.g. by clothes, contaminated tools, fruits and other plant parts, and at very low rate, by seed. PepMV is on the EPPO A2 List and is a regulated pathogen on tomato seeds in the EU. Containment of disease is achieved by sanitary measures, including seed disinfection (e.g. commercial bleach solution, 4) and seed testing. DAS-ELISA (1) is one of the established methods for reliable PepMV detection in plants and seeds.
Type of sample:
Leaves; Seeds
Target:
Pepino mosaic virus (PepMV)
Available extraction method:
For testing tomato, leaf samples are homogenized 1:20 (w/v) in extraction buffer «General» (Art. No. 110120). For seed testing, the following procedure is recommended: Slightly crush the seeds, e.g. with a hammer, 1:25 (w/v) in extraction buffer «General» (Art. No. 110120), then pre-soak them at least for 4 h (and up to overnight) at 4°C before homogenization (with HOMEX) and testing.
Available extraction method PDF:
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Scope:
The broad-spectrum reagents were made against a selection of PepMV isolates representing all major genotypes and geographic origins; i.e. Ch1 (US1 genotype), Ch2 (CH2 genotype) isolates (3), AZ-2 and TX-1 isolates (EU genotype) (5). The reagents specifically react with PepMV in DAS-ELISA; isolates of all strain groups of different geographic origin are recognized. The virus is detected in leaves and seeds. For testing tomato, leaf samples are homogenized 1:20 (w/v) in extraction buffer «General» (Art. No. 110120). For seed testing, the following procedure is recommended: Slightly crush the seeds, e.g. with a hammer, 1:25 (w/v) in extraction buffer «General», then pre-soak them at least for 4 h (and up to overnight) at 4°C before homogenization (with HOMEX) and testing.