Development of Real-Time RT-PCR Assay for Detection of <i>Prunus necrotic ringspot virus</i> in Fruit Trees

Marbot, Sophie; Salmon, Michel; Vendrame, Marina; Huwaert, Astrid; Kummert, J.; Dutrecq, O.; Lepoivre, Philippe

doi:10.1094/pdis.2003.87.11.1344

Cited by 28 publications

(16 citation statements)

References 25 publications

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“…Many research programs shifted from traditional methods (symptomatology, electron microscopy, and host differential reactions) to PCR-based or immunological tests when the latter were demonstrated to be more sensitive or specific (12,27,112,134). In limited cases, methods have been standardized among laboratories to ensure that comparisons between/among the groups were reliable (44).…”

Section: Comparison and Validation Of Current Microbial Forensic Idenmentioning

confidence: 99%

Plant Pathogen Forensics: Capabilities, Needs, and Recommendations

Fletcher

Bender

Budowle

et al. 2006

Microbiol Mol Biol Rev

161

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SUMMARY A biological attack on U.S. crops, rangelands, or forests could reduce yield and quality, erode consumer confidence, affect economic health and the environment, and possibly impact human nutrition and international relations. Preparedness for a crop bioterror event requires a strong national security plan that includes steps for microbial forensics and criminal attribution. However, U.S. crop producers, consultants, and agricultural scientists have traditionally focused primarily on strategies for prevention and management of diseases introduced naturally or unintentionally rather than on responding appropriately to an intentional pathogen introduction. We assess currently available information, technologies, and resources that were developed originally to ensure plant health but also could be utilized for postintroduction plant pathogen forensics. Recommendations for prioritization of efforts and resource expenditures needed to enhance our plant pathogen forensics capabilities are presented.

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Section: Comparison and Validation Of Current Microbial Forensic Idenmentioning

confidence: 99%

Plant Pathogen Forensics: Capabilities, Needs, and Recommendations

Fletcher

Bender

Budowle

et al. 2006

Microbiol Mol Biol Rev

161

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“…The Verso 1‐Step RT‐PCR was used as described earlier in separate assays with PDV‐ (Parakh et al. 1995) or PNRSV‐specific primers (Marbot et al. 2003) to test nine nucleic acid extracts of pollen collected from peach trees co‐infected with PDV and PNRSV.…”

Section: Methodsmentioning

confidence: 99%

Sensitive Detection of Viruses in Pollen Using Conventional and Real‐time Reverse Transcription‐polymerase Chain Reaction

Shiller¹,

Lebas²,

Horner

et al. 2010

Journal of Phytopathology

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Pollen is traded internationally as a source of germplasm and for pollination. Thirty-nine viruses and five viroids are known to be pollen transmitted. We investigated whether reverse transcription-polymerase chain reaction (RT-PCR) could be used to detect viruses reliably in pollen. Four extraction methods yielded nucleic acid in appropriate quantity and quality from Tobacco ringspot virus (TRSV)-infected pollen for RT-PCR amplification. One method, the RNeasy Ò Plant Mini Kit was used subsequently to extract nucleic acid of amplifiable quality from nine plant species, and pollen infected with three ilarvirus and two nepovirus species. A real-time TaqManÔ RT-PCR for the detection of TRSV was reliable and specific using 167 extracts of pollen from plants of Nicotiana glutinosa. The assay was highly sensitive, with extracts testing positive to a 10 )6 dilution, equivalent to a single pollen grain. This demonstrated that RT-PCR methods can detect virusinfected pollen reliably, sensitively and specifically. The possible application of these RT-PCR methods to replace current quarantine procedures without compromising biosecurity is discussed.

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“…However, such techniques are cumbersome and frequently lack sensitivity; therefore, other detection assays are generally used. Molecular detection assays based on molecular hybridisation or on reverse transcription polymerase chain reaction are also available (Rowhani et al, 1995;Hammond et al, 1999;Marbot et al, 2003;Sánchez-Navarro et al, 2005;reviewed in Hammond, 2011) and permit reliable detection of the virus. Molecular detection assays based on molecular hybridisation or on reverse transcription polymerase chain reaction are also available (Rowhani et al, 1995;Hammond et al, 1999;Marbot et al, 2003;Sánchez-Navarro et al, 2005;reviewed in Hammond, 2011) and permit reliable detection of the virus.…”

Section: Detection and Identification Of Prunus Necrotic Ringspot Virusmentioning

confidence: 99%

Scientific Opinion on the pest categorisation ofPrunus necrotic ringspot virus

Rossi¹

2014

EFSA Journal

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The Panel on Plant Health performed a pest categorisation of Prunus necrotic ringspot virus (PNRSV) for the European Union (EU) territory. PNRSV is a well-defined virus species of the genus Ilarvirus for which the entire genome sequence and molecular detection assays are available. It is transmitted by vegetative multiplication of infected hosts and also via seeds and pollen (both horizontally and vertically) in some of its hosts. PNRSV has a somewhat restricted natural host range, which contains Prunus spp., hops, roses and Rubus ellipticus (yellow Himalayan raspberry). It is listed on plants of Rubus for planting in Annex IIAI of Directive 2000/29EC, probably as a result of confusion with the closely related Apple mosaic virus. PNRSV is widely present in the EU, but there are no records on its regulated hosts. It is not expected to be affected by ecoclimatic conditions wherever its hosts are present, and it has the potential to establish in large parts of the EU territory. PNRSV can spread through efficient seed-and pollen-mediated transmission mechanisms and through the movement of vegetatively propagated plants for planting. However, the existence of efficient and widely adopted certification systems for Prunus spp. constitutes a limitation to PNRSV spread. Although the virus alone or when in mixed infection can cause significant diseases in some hosts, the actual impact of PNRSV appears to be limited.

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Development of Real-Time RT-PCR Assay for Detection of Prunus necrotic ringspot virus in Fruit Trees

Cited by 28 publications

References 25 publications

Plant Pathogen Forensics: Capabilities, Needs, and Recommendations

Plant Pathogen Forensics: Capabilities, Needs, and Recommendations

Sensitive Detection of Viruses in Pollen Using Conventional and Real‐time Reverse Transcription‐polymerase Chain Reaction

Scientific Opinion on the pest categorisation ofPrunus necrotic ringspot virus

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