Improved real-time PCR assay for detection of the quarantine potato pathogen, Synchytrium endobioticum, in zonal centrifuge extracts from soil and in plants

Gent-Pelzer, M.P.E. van; Krijger, M.C.; Bonants, P.J.M.

doi:10.1007/s10658-009-9522-3

Cited by 38 publications

(28 citation statements)

References 13 publications

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“…; Van Gent‐Pelzer et al. ). Alternatively, sieving–centrifugation procedures before extraction can be utilized to concentrate the pathogen from larger samples and increase sampling representativeness (Pavón et al.…”

Section: Detection Of Soilborne Fungi and Oomycetesmentioning

confidence: 99%

Use of QuantitativePCRDetection Methods to Study Biocontrol Agents and Phytopathogenic Fungi and Oomycetes in Environmental Samples

Sanzani

Nicosia

Faedda³

et al. 2013

Journal of Phytopathology

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Quantitative polymerase chain reaction (qPCR) is a versatile technique for the accurate, sensitive, reliable and high‐throughput detection and quantification of target DNA in various environmental samples, and in recent years, it has greatly contributed to the advancement of knowledge in the plant pathology field. Indeed, this technique is ideal to evaluate inoculum threshold levels and to study the epidemiology, biology and ecology of phytopathogenic fungi and oomycetes, thus opening up new research opportunities to investigate host–pathogen interactions and to address tasks related to quarantine, eradication and biosecurity. Moreover, it can be a useful tool in breeding programs. The present review analyses the most relevant applications of qPCR for the detection and quantification of filamentous fungi and oomycetes within host tissues and in soil, air and water, along with brief paragraphs focusing on new application fields such as the detection and quantification of mycotoxigenic fungi and biocontrol agents. The high potentiality of qPCR for present and future applications is highlighted together with a critical analysis of major drawbacks that need to be corrected to definitively confirm it as a preferential routine quantitative detection method.

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Section: Detection Of Soilborne Fungi and Oomycetesmentioning

confidence: 99%

Use of QuantitativePCRDetection Methods to Study Biocontrol Agents and Phytopathogenic Fungi and Oomycetes in Environmental Samples

Sanzani

Nicosia

Faedda³

et al. 2013

Journal of Phytopathology

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“…The identity of summer sporangia or resting spores observed under the microscope can be confirmed by conventional PCR (Lévesque et al ., ; van den Boogert et al ., ), and real‐time PCR (van Gent‐Pelzer et al ., ; Smith et al ., ). An international test performance study (TPS) was organized under the Euphresco SENDO project to generate validation data for three molecular S. endobioticum detection and identification tests (van den Boogert et al ., ; van Gent‐Pelzer et al ., ; Bonants et al ., ). Two TPS rounds were organized focusing on different test matrices: wart material and resting spore suspensions.…”

Section: Identificationmentioning

confidence: 97%

“…The real‐time PCR of van Gent‐Pelzer et al . () is based on ITS2. The real‐time PCR of Smith et al .…”

Section: Identificationmentioning

confidence: 99%

PM 7/28 (2) Synchytrium endobioticum

2017

EPPO Bulletin

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“…Because inhibition of PCR inhibitions often occurs in organic rich soil media before extracting total DNA, a method for the prior assessment of DNA quality might be important, despite recent improvements in DNA extraction methods. Alternatively, another set of primers and probe of the internal amplification control might be added to each PCR reaction to ascertain any inhibition of amplification, and false negative results should be reassessed when the experimental budget allows such expenditures (Klerks et al 2004;van Gent-Pelzer et al 2010).…”

Section: Development Of Real-time Pcr Assay Using Taqman Probementioning

confidence: 99%

Development of real-time PCR assay using TaqMan probe for detection and quantification of Rosellinia necatrix in plant and soil

2012

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We developed real-time PCR assays using TaqMan probes to detect and quantify Rosellinia necatrix, the causal agent of white root rot in many plant species. Two sets of PCR primers and TaqMan probe indicated that their detection limits could be as low as 1 fg of template DNA. Using the real-time PCR assays with the TaqMan probes, we were able to quantify R. necatrix DNA in naturally diseased roots of Japanese pear and in artificially infested soil samples. Although the new assays were inadequate for use with naturally infested soil samples, nested PCR procedures improved the detectability of the new assays.

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Improved real-time PCR assay for detection of the quarantine potato pathogen, Synchytrium endobioticum, in zonal centrifuge extracts from soil and in plants

Abstract: Real-time PCR was used for quantitative detection of the potato pathogen, Synchytrium endobioticum, in different substrates: zonal centrifuge extracts, warts and different plant parts of potato.

Cited by 38 publications

References 13 publications

Use of QuantitativePCRDetection Methods to Study Biocontrol Agents and Phytopathogenic Fungi and Oomycetes in Environmental Samples

Use of QuantitativePCRDetection Methods to Study Biocontrol Agents and Phytopathogenic Fungi and Oomycetes in Environmental Samples

PM 7/28 (2) Synchytrium endobioticum

Development of real-time PCR assay using TaqMan probe for detection and quantification of Rosellinia necatrix in plant and soil

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