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

Fajardo, T. V. M.; Nickel, Osmar

doi:10.1590/0103-8478cr20131629

Cited by 6 publications

(4 citation statements)

References 8 publications

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“…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.…”

Section: Discussionmentioning

confidence: 99%

“…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.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2015

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Clavibacter michiganensis subsp. michiganensis, Pepino mosaic virus, and Mexican papita viroid are three economically important pathogens that infect tomato crops. In this work, a polyprobe (poly-3) was developed and evaluated for the simultaneous detection of these pathogens in tomato plants by non-isotopic molecular hybridization. The endpoint detection limit of the poly-3 with C. michiganensis subsp. michiganensis cell cultures was 10 6 CFU/ml. No differences were found in terms of the sensitivity and specificity when the individual riboprobes were compared to the poly-3 for the detection of the three pathogens. The analysis of 80 tomato field samples by the poly-3 and RT-PCR techniques rendered the same number of positive samples for each pathogen. As far as we know this is the first time that three pathogens with very different life cycle styles (bacteria, virus and viroid) are simultaneously detected in a single assay. The possibility of using this poly-3 technology for the routine diagnosis of field tomato samples is discussed.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2015

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“…Polyprobes have been applied successfully for the detection of an array of combinations of plant viruses and/or viroids affecting different crops ( Table 1 ). Polyprobes have been developed and used to detect four viroids affecting citrus trees ( Cohen et al, 2006 ), the six main viruses infecting tomato ( Aparicio et al, 2009 ), six viruses affecting pome and stone fruits ( Lin et al, 2011 ), four viroids affecting grapevine ( Zhang et al, 2012 ), eight viroids affecting ornamentals and vegetables ( Torchetti et al, 2012 ), eight viroids infecting coleus plants ( Jiang et al, 2013 ), and four viruses of apple and pear trees ( Fajardo and Nickel, 2014 ). Remarkably, this technology has great potential for simultaneous multipathogen detection.…”

Section: Molecular Hybridizationmentioning

confidence: 99%

Recent Advances on the Multiplex Molecular Detection of Plant Viruses and Viroids

2018

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Plant viruses are still one of the main contributors to economic losses in agriculture. It has been estimated that plant viruses can cause as much as 50 billion euros loss worldwide, per year. This situation may be worsened by recent climate change events and the associated changes in disease epidemiology. Reliable and early detection methods are still one of the main and most effective actions to develop control strategies for plant viral diseases. During the last years, considerable progress has been made to develop tools with high specificity and low detection limits for use in the detection of these plant pathogens. Time and cost reductions have been some of the main objectives pursued during the last few years as these increase their feasibility for routine use. Among other strategies, these objectives can be achieved by the simultaneous detection and (or) identification of several viruses in a single assay. Nucleic acid-based detection techniques are especially suitable for this purpose. Polyvalent detection has allowed the detection of multiple plant viruses at the genus level. Multiplexing RT polymerase chain reaction (PCR) has been optimized for the simultaneous detection of more than 10 plant viruses/viroids. In this short review, we provide an update on the progress made during the last decade on techniques such as multiplex PCR, polyvalent PCR, non-isotopic molecular hybridization techniques, real-time PCR, and array technologies to allow simultaneous detection of multiple plant viruses. Also, the potential and benefits of the powerful new technique of deep sequencing/next-generation sequencing are described.

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“…A caracterização molecular, em que um isolado viral ou suas variantes e espécies podem ser diferenciados com base na sequência genômica, é uma fer-ramenta importante para classificar as espécies virais (TORRICO et al, 2016). Para a detecção de vírus em morangueiro, as técnicas mais utilizadas são a RT-PCR (Reverse Transcription Polymerase Chain Reaction) e o teste ELISA (Enzyme Linked Immunosorbent Assay) (MAR- TIN & TZANETAKIS, 2006), embora outros métodos, como a PCR (Polymerase Chain Reaction), q-PCR (PCR quantitativo ou em tempo real) e o dot-ELISA, sejam empregados na detecção de outras viroses (FAJARDO & NICKEL, 2014).…”

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Detecção molecular e análise filogenética da sequência parcial do gene da proteína do capsídeo do vírus da faixa das nervuras do morangueiro

Puttkammer

Zappelini

Klabunde

et al. 2021

RAC

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O morango cultivado (Fragaria x ananassa Duch. (Rosaceae) é um híbrido originado pelo cruzamento das espécies americanas Fragaria chiloensis e Fragaria virginiana e pertence à família Rosaceae. O morangueiro possui reprodução vegetativa e, por isso, é comum o acúmulo de viroses e outras doenças de difícil controle. Uma das quatro viroses mais importantes na cultura do morango é causada pelo vírus da faixa das nervuras (Strawberry vein banding virus – SVBV), que é transmitido no campo por afídeos, de maneira semipersistente. Para melhorar o conhecimento genômico, são recomendadas técnicas moleculares e a classificação de isolados de SVBV. Um dos determinantes primários da transmissibilidade e especificidade por afídeos é a proteína do capsídeo, que possui importância crítica para o estabelecimento da infecção. Neste trabalho, foi sequenciada parcialmente a proteína do capsídeo do gene do SVBV de um isolado alemão, inoculado em morangueiros e mantido em casa de vegetação no Brasil, por mais de dez anos. Foi realizada a análise filogenética comparando as sequências contidas no GenBank com o objetivo de elucidar as relações evolutivas nesta espécie. As análises filogenéticas mostraram que a sequência do isolado está mais próxima dos isolados dos EUA e Egito. Estes resultados contribuem para a melhor elucidação dos mecanismos de evolução do vírus e do patossistema em questão.

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

Cited by 6 publications

References 8 publications

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

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

Recent Advances on the Multiplex Molecular Detection of Plant Viruses and Viroids

Detecção molecular e análise filogenética da sequência parcial do gene da proteína do capsídeo do vírus da faixa das nervuras do morangueiro

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