Simultaneous detection of six RNA plant viruses affecting tomato crops using a single digoxigenin-labelled polyprobe

Abstract

A polyprobe for the simultaneous detection by non-isotopic molecular hybridisation has been developed to detect any of the following six viruses causing important economic losses in tomato crops: Tomato spotted wilt virus, Tomato mosaic virus, Pepino mosaic virus, Cucumber mosaic virus, Potato Y virus and Parietaria mottle virus. The polyprobe detected all six viruses with similar sensitivity to that obtained using individual riboprobes. In addition, we evaluated the possible use of the tissue-printing as a sample preparation technique applied to routine diagnosis of tomato plants with the polyprobe.

Simultaneous detection of four viruses affecting apple and pear by molecular hybridization using a polyprobe

“…For both aspects it was found that the evaluated riboprobe did not compromise the virus detection of assayed samples (Table 1). Supporting the relevance of the present results for pathogen detection using molecular probes, in many studies, molecular non-radioactive hybridization for simultaneous detection of different viruses and/or viroids using a single probe or polyprobe containing different viral and/or viroidal nucleotide sequences has been developed, and successfully used for pathogen diagnosis (HERRANZ et al, 2005;APARICIO et al, 2009;FAJARDO et al, 2012;PEIRÓ et al, 2012).…”

“…This method permits the simultaneous detection of different viruses in one test with detection limit similar to that obtained by ELISA. This technology has proved to be an effi cient and cheap method for the detection of the main viruses affecting stone fruits (HERRANZ et al, 2005;PEIRÓ et al, 2012), tomatoes (APARICIO et al, 2009) and grapevines (FAJARDO et al, 2012). The objective of this study was to develop a polyprobe with the capacity to simultaneously detect four main viruses affecting apple and pear plants.…”

Simultaneous detection of Clavibacter michiganensis subsp. michiganensis, Pepino mosaic virus and Mexican papita viroid by non-radioactive molecular hybridization using a unique polyprobe

“…The development of techniques to detect simultaneously the most important pathogens threatening a specific crop is desirable, because it reduces time and associated costs of producing pathogen freematerial (James et al 2010). Simultaneous detection of several viruses and/or viroids in a single test using the NiMH technology is a reliable diagnostic procedure (Saldarelli et al 1996;Sanchez-Navarro et al 1999;Saade et al 2000;Ivars et al 2004;Herranz et al 2005;Cohen et al 2006;Aparicio et al 2009;Lin et al 2011;Peiró et al 2012;Fajardo and Nickel 2014). However, as far as we know, no simultaneous detection of a bacterium, virus and viroid by NiMH has been yet reported.…”

“…The NiHM technology using polyprobes has permitted the simultaneous detection of four or six viroids (Cohen et al 2006;Lin et al 2011), six viruses (Herranz et al 2005;Aparicio et al 2009) or eight viruses plus two viroids (Peiró et al 2012). Here we present data showing the poly-3 probe allows the simultaneous detection of three pathogens (bacterium, virus and viroid) with the same detection limit and specificity compared to individual riboprobes.…”

Development of a polyprobe for the simultaneous detection of four grapevine viroids in grapevine plants

“…In recent years, non-isotopic molecular hybridization techniques have been used for the simultaneous detection of multiple plant viroids and viruses. This approach has become an attractive method for routine diagnosis due to advantages of low-cost and practicality (Herranz et al 2005;Cohen et al 2006;Aparicio et al 2008;Lin et al 2011).…”