Implementation of an artificial reaction control in a TaqMan method for PCR detection of Ralstonia solanacearum race 3 biovar 2

Smith, Donna S.; Boer, S. H. De

doi:10.1007/s10658-008-9427-6

Cited by 11 publications

(16 citation statements)

References 18 publications

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“…The modified TaqMan assay procedure, developed in this investigation, successfully detected Rs race 3 bv. 2 in infected asymptomatic tomato stems and leaves of tomato, as well as in potato tubers and stems (Smith and De Boer 2009).…”

Section: Real-time Polymerase Chain Reactionmentioning

confidence: 96%

Detection of Bacterial and Phytoplasmal Pathogens

Narayanasamy¹

2010

Microbial Plant Pathogens-Detection and Disease Diagnosis:

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Bacterial pathogens classified as prokaryotes are much simpler in structure and smaller in size and they have less complex structural features compared to fungal plant pathogens. As the morphological characteristics of bacterial cells are less variable, biological, biochemical, physiological, immunological and genomic characteristics have to be determined for reliable identification and meaningful classification of bacterial pathogens. Detection and identification of bacterial plant pathogens present in whole plants and in propagative plant materials have been possible by employing isolation on cultural media and metabolic fingerprinting methods. But they are labor-intensive and require long time. Results often are inconclusive. Isozyme analysis, direct colony thin layer chromatography and gel electrophoresis techniques have been successfully applied for the detection of some bacterial pathogens. Immunoassays and nucleic acid-based assays have become widely accepted techniques, providing more sensitive and specific detection and quantification of bacterial pathogens affecting a wide range of host plant species. However, the major limitation of the molecular techniques is their inability to discriminate living cells from the dead ones. Attempts have been made to overcome this limiting factor by using DNA binding dyes and estimating pathogen RNAs as an indicator of cell viability. The diagnostic methods have both merits and demerits and hence, appropriate method has to be selected based on cost-effectiveness and possibility of obtaining reliable results rapidly to suit the requirements of the investigation concerned.Phytoplasmas are cell wall-less, nonculturable bacteria belonging to Mollicutes. Lack of cultural characteristics has made obligatory to study the morphological characteristics of phytoplasmas present in the phloem cells of infected plant hosts by observing under electron microscopes. As most of the phytoplasmas look alike in the ultrathin sections, the morphological characters have no diagnostic value. Histochemical methods using DNA binding dyes have been employed to localize the phytoplasmas in the host cells. Application of immunoassays and nucleic acidbased techniques has been effective in providing information for the identification and differentiation of phytoplasams. Polyclonal and monoclonal antibodies have been generated for detection of phytoplasmas infecting several plant species. Universal and species-specific primers and probes have been designed based on

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Section: Real-time Polymerase Chain Reactionmentioning

confidence: 96%

Detection of Bacterial and Phytoplasmal Pathogens

Narayanasamy¹

2010

Microbial Plant Pathogens-Detection and Disease Diagnosis:

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“…The real-time TaqMan PCR assay of Smith and De Boer (2009) with internal control was used to compare with a real-time PCR assay originally described by Ozakman and Schaad (2003). Both assays are used for detecting R. solanacearum R3bv2 in asymptomatic potato tubers.…”

Section: Detection By Real-time Taqman Pcr Assaysmentioning

confidence: 99%

“…However, the two assays differed in sensitivity (Table 3). The assay based on TaqMan chemistries with internal control (Pastrik et al 2000;Smith and De Boer 2009) showed the highest sensitivity (Tables 3 and 4, gel images see supporting materials), detecting spiked concentration of 68, 86 and 120 CFU ml À1 for R3bv2 strains NCPPB339, B0167 and NCPPB909, respectively. The detection limit for 15 other R3bv2 strains was at a similar range.…”

Section: Specific Assays For Detection Of Ralstonia Solanacearum R3bvmentioning

confidence: 99%

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

Nie

Hammill

et al. 2014

J Appl Microbiol

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Aims: To determine the reliable combination of protocols for specific detection and identification of R. solanacearum race 3 biovar 2 (R3bv2) through a comprehensive comparison among currently available techniques. Methods and Results: Sensitivity and specificity of the conventional isolation, bioassay, serological assays, conventional and real-time PCR and multiplex PCR were assessed for the detection of 25 strains of R. solanacearum biovars 1, 2 and 3 (Phylotypes I, II, III and IV) in spiked potato saps. Results indicated that all assays evaluated varied in complexity and sensitivity and should be applied strategically in indexing schemes to maximize efficiency of testing without compromising accuracy of the results. Conclusions: The TaqMan PCR assay, with an internal reaction control and confirmation by melting curve and electrophoretic analysis, achieved best sensitivity at 10 2 -10 3 CFU ml À1 for all eighteen strains of R. solanacearumR3bv2. Selective enrichment on mSMSA medium plates enhanced the detection sensitivity up to 10-100 CFU ml À1 for the conventional PCR-based assays.Significance and Impact of the Study: This is the first time nine different assays were compared side by side for their sensitivity and specificity in detection and identification of R. solanacearum R3bv2. The data accumulated here will provide basis for regulatory applications for low level detection and rapid identification of latently infections caused by R. solanacearum R3bv2.

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“…Grover et al . () used MDA‐PCR to detect ultra‐low populations; Smith and de Boer () developed an improved TaqMan method for PCR; Poussier et al . (), Alvarez Restrepo et al .…”

Section: Specific Groups Of Exotic Pathogensmentioning

confidence: 99%

A review of pest surveillance techniques for detecting quarantine pests in Europe

Augustin¹,

Boonham

Kogel³

et al. 2012

EPPO Bulletin

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This paper provides reviews of the most commonly used methods to detect plant pests belonging to groups of invasive organisms with high economic relevance, including Coleoptera (bark beetles, flathead borers, leaf beetles, longhorn beetles, weevils), Diptera (cone and seed flies, fruit flies), Homoptera (aphids, leafhoppers and psyllids, whiteflies), Lepidoptera (moths and butterflies), Thysanoptera (thrips), bacteria (potato brown rot Ralstonia solanacearum) and fungi (pitch canker disease Gibberella circinata, brown rot disease Monilinia fructicola). Future perspectives in detection methods are discussed, with particular reference to the considerable increase in the volume, commodity type and origins of trade in plant material from third countries, the introduction of new crops, the continuous expansion of the EU with new border countries being added, and the impact of climate change affecting the geographical boundaries of pests and their vectors. (Résumé d'auteur

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Implementation of an artificial reaction control in a TaqMan method for PCR detection of Ralstonia solanacearum race 3 biovar 2

Cited by 11 publications

References 18 publications

Detection of Bacterial and Phytoplasmal Pathogens

Detection of Bacterial and Phytoplasmal Pathogens

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

A review of pest surveillance techniques for detecting quarantine pests in Europe

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