Since 1988, viroids have been occasionally detected in samples of tomato (Lycopersicon esculentum) originating both in the Netherlands and other countries. Infected plants showed chlorosis, bronzing, leaf distortion and growth reduction. Initial diagnosis of these viroids was by return-polyacrylamide gel electrophoresis, which did not allow a further identification. This paper reports the identification of these viroids by reverse transcription-polymerase chain reaction and sequence analysis. Three known viroids of tomato, i.e. Citrus exocortis viroid; Potato spindle tuber viroid and Tomato chlorotic dwarf viroid were identified. In addition, six isolates were identified as Columnea latent viroid, a viroid so far only detected in some ornamental species. Like the isolates previously isolated from ornamental species, the isolates from tomato share genetic characteristics of both the genera Hostuviroid and Pospiviroid. The biological characteristics of all four viroids, especially their potential effects on both potato (Solanum tuberosum) and tomato, stress the need for reconsideration of their phytosanitary risks.

No abstract

A multiplex quantitative PCR (qPCR) was developed and evaluated for the simultaneous detection of Salmonella spp., S. enterica serovar Typhimurium and S. enterica serovar Enteritidis in various (food) matrices. Early and fast detection of these pathogens facilitates effective intervention and prevents further distribution of contaminated food products on the market. Three primer and probe sets were designed to target the invA gene, the STM4200 gene, and the SEN1392 gene to detect and differentiate Salmonella spp., S. Typhimurium, and S. Enteritidis, respectively. The multiplex qPCR targeting these three genes was optimized for efficiency and linearity. By testing 225 Salmonella isolates and 34 non-Salmonella isolates from various sources the inclusivity and exclusivity were determined. The inclusivity of the multiplex qPCR was 100% for all Salmonella isolates, including 72 S. Typhimurium isolates, and 53 S. Enteritidis isolates. The exclusivity for Salmonella spp., S. Typhimurium, and S. Enteritidis was 100%, 94.6%, and 100%, respectively. No positive results were reported for non-Salmonella isolates. The limit of detection (LOD) for the qPCR was determined for the matrices poultry, minced meat, egg, herbs/spices, powdered milk, fish, animal feed, boot-socks with chicken feces and chicken down. LOD values for qPCR and the conventional culture methods were similar, except for the matrix boot-socks and down, for which the LOD for the conventional culture methods performed better than the qPCR method. In conclusion, the multiplex qPCR assay developed allows for rapid screening of Salmonella spp., S. Typhimurium, and S. Enteritidis in various (food) matrices.

No abstract

No abstract