A comprehensive comparison of assays for detection and identification of <i>Ralstonia solanacearum</i>
 race 3 biovar 2

Li, X.; Nie, J.; Hammill, Desmond L; Smith, Donna S.; Xu, Huimin; Boer, S. H. De

doi:10.1111/jam.12585

Cited by 16 publications

(9 citation statements)

References 20 publications

(58 reference statements)

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“…Gutarra et al (2017) describe their research on the diversity of the species and key information for the species/strains/biovar/phylotype/sequevar identification. Li et al (2014) provide details on sensitivity and specificity of the methods used for comparisons of the detection and identification assays in the laboratory for testing the potato samples for the presence of the causal agent of potato brown rot.…”

Section: Laboratory Testing and Pest Identificationmentioning

confidence: 99%

Pest survey card on potato brown rot, Ralstonia solanacearum

Gaag¹,

Camilleri²,

Diakaki³

et al. 2019

EFS3

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This pest survey card was prepared in the context of the mandate on plant pest surveillance (EFSA-Q-2017-00831), upon request by the European Commission. The purpose of this document is to assist the Member States in planning annual survey activities of quarantine organisms using a statistically sound and risk-based pest survey approach, in line with the current international standards. The data requirements for such activity include the pest distribution, its host range, its biology, risk factors as well as available detection and identification methods. This document is part of a toolkit that consists of pest-specific documents, such as the pest survey cards and generic documents relevant for all pests to be surveyed, including, the general survey guidelines and statistical software such as RiBESS+.

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Section: Laboratory Testing and Pest Identificationmentioning

confidence: 99%

Pest survey card on potato brown rot, Ralstonia solanacearum

Gaag¹,

Camilleri²,

Diakaki³

et al. 2019

EFS3

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“…The detection limit for R. solanacearum with immunofluorescence, for example, is about 10 4 to 10 5 cfu mL À1 in pellet extract (Janse, 1988;Elphinstone et al, 1996;Vreeburg et al, 2016;van Vaerenbergh et al, 2017), while the detection limit of a pathogenicity test of R. solanacearum (referred to as race 3, biovar 2 in articles) in tomato plants is between 2 9 10 2 and 10 5 cfu mL À1 (Janse, 1988;Elphinstone et al, 1996;Li et al, 2014). The detection limit for R. solanacearum with immunofluorescence, for example, is about 10 4 to 10 5 cfu mL À1 in pellet extract (Janse, 1988;Elphinstone et al, 1996;Vreeburg et al, 2016;van Vaerenbergh et al, 2017), while the detection limit of a pathogenicity test of R. solanacearum (referred to as race 3, biovar 2 in articles) in tomato plants is between 2 9 10 2 and 10 5 cfu mL À1 (Janse, 1988;Elphinstone et al, 1996;Li et al, 2014).…”

Section: Potato Lot Ct Valuementioning

confidence: 99%

“…An infection with potato brown rot or potato ring rot can be verified by immunofluorescence and should be confirmed with a pathogenicity test. The detection limit for R. solanacearum with immunofluorescence, for example, is about 10 4 to 10 5 cfu mL À1 in pellet extract (Janse, 1988;Elphinstone et al, 1996;Vreeburg et al, 2016;van Vaerenbergh et al, 2017), while the detection limit of a pathogenicity test of R. solanacearum (referred to as race 3, biovar 2 in articles) in tomato plants is between 2 9 10 2 and 10 5 cfu mL À1 (Janse, 1988;Elphinstone et al, 1996;Li et al, 2014). For C. michiganensis subsp.…”

Section: Potato Lot Ct Valuementioning

confidence: 99%

Validation of four real‐time TaqMan PCRs for the detection of Ralstonia solanacearum and/or Ralstonia pseudosolanacearum and/or Clavibacter michiganensis subsp. sepedonicus in potato tubers using a statistical regression approach

Vreeburg

Zendman²,

Pol³

et al. 2018

EPPO Bulletin

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A new DNA extraction method and a new multiplex real‐time TaqMan PCR test for detection of Ralstonia solanacearum, Ralstonia pseudosolanacearum and Clavibacter michiganensis subsp. sepedonicus in asymptomatic potato tubers are presented. This new multiplex PCR and three published TaqMan PCRs for detection of R. solanacearum and/or R. pseudosolanacearum and/or R. syzygii spp. and/or C. michiganensis subsp. sepedonicus were validated using linear regression analysis for estimating the Ct values and its variation at 5 × 103 bacteria mL−1. The three published PCRs that have been validated are Massart et al. (2014, detecting R. solanacearum and C. michiganensis subsp. sepedonicus), Weller et al. (1999, detecting R. solanacearum, R. pseudosolanacearum and R. syzygii spp.) and Gudmestad et al. (2009, detecting C. michiganensis subsp. sepedonicus). All tested PCRs were fit for purpose for their target organisms. The PCR tests have different target genes, allowing one of the sets to be used as first screening test and another as second screening test for the detection of R. solanacearum and/or R. pseudosolanacearum and/or C. michiganensis subsp. sepedonicus in asymptomatic potato tubers.

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“…General protocols that rely on the evolution of PCR techniques are also available, such as qPCR (Weller et al, 2000; Ozakman and Schaad, 2003; Smith and De Boer, 2009; Inoue and Nakaho, 2014) or LAMP PCR (Lenarcic et al, 2014). Specific protocols have been designed to target a particular group of strains or ecotypes of the Rssc, such as brown rot strains (Fegan et al, 1998; Weller et al, 2000; Ozakman and Schaad, 2003; Smith and De Boer, 2009; Kubota et al, 2011; Ha et al, 2012; Li et al, 2014; Kubota and Jenkins, 2015; Stulberg et al, 2015); Moko strains (Prior and Fegan, 2005a; Cellier et al, 2015); and Blood Disease Bacterium (BDB) strains (Kubota et al, 2011). These diagnostic methods are primarily limited to the detection of strains associated with the brown rot ecotype (Li et al, 2014); however, the Rssc presents a wide diversity of strains.…”

Section: Introductionmentioning

confidence: 99%

Tube-Wise Diagnostic Microarray for the Multiplex Characterization of the Complex Plant Pathogen Ralstonia solanacearum

Cellier¹,

Arribat

Chiroleu

et al. 2017

Front. Plant Sci.

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Ralstonia solanacearum is a well-known agricultural and ecological threat worldwide. The complexity of the R. solanacearum species complex (Rssc) represents a challenge for the accurate characterization of epidemiological strains by official services and research laboratories. The majority of protocols only focus on a narrow range of strains; however, this species complex includes strains that represent major constraints and are under strict regulation. The main drawback associated with the current methods of detecting and characterizing Rssc strains is their reliance on combining different protocols to properly characterize the strains at the ecotype level, which require time and money. Therefore, we used microarray technology (ArrayTube) to develop a standard protocol, which characterizes 17 major groups of interest in the Rssc, in a single multiplex reaction. These 17 majors groups are linked with a phylogenetic assignation (phylotypes, sequevars), but also with an ecotype assignation associated with a range of hosts (e.g., brown rot, Moko). Probes were designed with a 50-mer length constraint and thoroughly evaluated for any flaws or secondary structures. The strains are characterized based on a DNA extraction from pure culture. Validation data showed strong intra-repeatability, inter-repeatability, and reproducibility as well as good specificity. A hierarchical analysis of the probe groups is suitable for an accurate characterization. Compared with single marker detection tests, the method described in this paper addresses efficiently the issue of combining several tests by testing a large number of phylogenetic markers in a single reaction assay. This custom microarray (RsscAT) represents a significant improvement in the epidemiological monitoring of Rssc strains worldwide, and it has the potential to provide insights for phylogenetic incongruence of Rssc strains based on the host of isolation and may be used to indicate potentially emergent strains.

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A comprehensive comparison of assays for detection and identification of Ralstonia solanacearum race 3 biovar 2

Cited by 16 publications

References 20 publications

Pest survey card on potato brown rot, Ralstonia solanacearum

Pest survey card on potato brown rot, Ralstonia solanacearum

Validation of four real‐time TaqMan PCRs for the detection of Ralstonia solanacearum and/or Ralstonia pseudosolanacearum and/or Clavibacter michiganensis subsp. sepedonicus in potato tubers using a statistical regression approach

Tube-Wise Diagnostic Microarray for the Multiplex Characterization of the Complex Plant Pathogen Ralstonia solanacearum

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