Sensitive detection and discrimination of different Pepino mosaic virus genotypes

• Published in: Phytopathology Vol. 100; no. 6; p. S81
• Main Authors: Mehle, N, Gutierrez-Aguirre, I, Prezelj, N, Delic, D, Vidic, U, Kramberger, P, Ravnikar, M
• Format: Journal Article
• Language: English
• Published: 01-06-2010
• Subjects: Lycopersicon esculentum; Pepino mosaic virus
• ISSN: 0031-949X

We developed quantitative real time PCR (RT-qPCR) assays for the detection and discremination of presently circulating PepMV (Pepino mosaic virus) genotypes. The following genotype combinations, European tomato-Peruvian, Ch2, and US1, were successfully distinguished within the tested isolates. One-step RT-qPCR detected PepMV European tomato genotype particles at least two orders of magnitude more sensitively than ELISA. The method detected as little as one naturally infected seed among 5000 uninfected seeds. The developed RT-qPCR assay will enable detailed studies on the demographic distribution of the circulating PepMV genotypes. At the same time, the disposability of more than one gene target for the detection of PepMV RNA, will increase the reliability of the virus detection in samples with low expected virus concentration, such as latent infections or irrigation waters. It is suspected that PepMV can be present in the environment at concentrations, which despite being too low to be detected by conventional detection methods, they can still constitute a risk of potential infection. We have experimentally confirmed that PepMV remained infective in water for up to three weeks. We are investigating the potential of monolithic chromatographic supports (CIM Convective Interaction Media) for concentration of PepMV. The combination of both technologies (CIM for concentration and qPCR for detection) will allow detection of extremely low concentrations of PepMV from environmental waters.