Development of a polyprobe to detect six viroids of pome and stone fruit trees

Determination of Major Viral and Sub Viral Pathogens Incidence in Apple Orchards in Himachal Pradesh

“…In this study molecular methods for viral and sub viral pathogen detection (RT-PCR and NASH) for early diagnostics was optimized, which helps in effective diagnostics both in vitro and in vivo of apple orchards and nurseries. Also results suggests the need for development of reliable, fast and cost effective molecular method which detects these major pathogens in one reaction like multiplex reverse transcription-polymerase chain reaction [11] or the use of polyvalent probe [9]. Three methods were described for detection of viral pathogens and two for subviral pathogen detection.…”

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.…”

“…Another approach uses a unique riboprobe, called polyprobe, which contains fused-in-tandem partial nucleic acid sequences of the target pathogens (Herranz et al 2005). Polyvalent detection using a unique polyprobe has been successfully applied to the detection of different plant viruses infecting stone fruits (Herranz et al 2005;Peiró et al 2012;Fajardo and Nickel 2014), tomato ) and different viroids that infect citrus (Cohen et al 2006) pome and stone fruit trees (Lin et al 2011), grapevine (Zhang et al 2012) and ornamentals and vegetables (Torchetti et al 2012). Moreover, the use of the polyprobe technology by NiMH has been proven to allow the detection of virus and viroids in a single assay.…”

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).…”