A Combination of Baiting and Different PCR Formats, Including Measurement of Real-Time Quantitative Fluorescence, For the Detection of Phytophthora fragariae in Strawberry Plants

Bonants, P.J.M.; Gent-Pelzer, M.P.E. van; Hooftman, R.; Cooke, D. E. L.; Guy, D. C.; Duncan, J. M.

doi:10.1023/b:ejpp.0000041551.26970.0e

Cited by 41 publications

(30 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Detection of quarantine fungi requires highly sensitive and specific methods. Real‐time TaqMan PCR assays meet these requirements (Bonants et al., 2004; Schena et al., 2004; van den Boogert et al., 2005). Additional advantages are a closed tube system in which contamination is minimized and no gel electrophoresis, which saves time and provides the possibility for high throughput screening.…”

Section: Introductionmentioning

confidence: 99%

A TaqMan PCR Method for Routine Diagnosis of the Quarantine Fungus Guignardia citricarpa on Citrus Fruit

Gent-Pelzer¹,

Brouwershaven²,

Kox³

et al. 2007

Journal of Phytopathology

View full text Add to dashboard Cite

With respect to disease risk for the quarantine fungus Guignardia citricarpa on citrus fruit an accurate diagnosis for routine analysis is required. Also, when inspections have to be performed on imported citrus fruits, a fast detection method is urgently needed. A fast automated DNA extraction method based on magnetic beads combined with a real-time PCR assay was optimized to improve and advance the routine diagnosis of citrus black spot disease. Real-time PCR was used for detection of the pathogen G. citricarpa in planta. A specific primer/TaqMan probe combination that discriminates between G. citricarpa and the harmless citrus endophyte Guignardia mangiferae, was designed based on the internal transcribed spacer region of the multi-copy rDNA gene. Co-amplification of target DNA along with an internal competitor DNA fragment made the diagnostic assay more reliable to check for false negatives. The real-time PCR was specific, since no cross reaction was observed with a series of citrus pathogens and related species. The diagnostic assay was performed on lesions dissected from imported diseased oranges. Comparison between the conventional PCR and the real-time PCR methods showed that the TaqMan method was more sensitive.

show abstract

Section: Introductionmentioning

confidence: 99%

A TaqMan PCR Method for Routine Diagnosis of the Quarantine Fungus Guignardia citricarpa on Citrus Fruit

Gent-Pelzer¹,

Brouwershaven²,

Kox³

et al. 2007

Journal of Phytopathology

View full text Add to dashboard Cite

show abstract

“…In particular, real-time PCR methods have the advantages of speed, accuracy, and sensitivity over other methods (7,28,30,43) and can be closedtube systems involving no postamplification steps. TaqMan (11,16) is the most widely used real-time PCR system, and assays using this detection chemistry have been described for a range of plant pathogens (3,4,14,39,38,40), including assays for the detection of Phytophthora ramorum both in the laboratory (8,12,34) and in the field (33). A single round of real-time PCR has been found to be sufficiently sensitive to achieve the detection of 10 to 100 fg P. ramorum DNA in plant material (8,33).…”

mentioning

confidence: 99%

Faster, Simpler, More-Specific Methods for Improved Molecular Detection of Phytophthora ramorum in the Field

Tomlinson¹,

Barker²,

Boonham³

2007

Appl Environ Microbiol

138

View full text Add to dashboard Cite

Phytophthora ramorum is the causal agent of sudden oak death. The pathogen also affects a wide range of tree, shrub, and herbaceous species in natural and landscaped environments as well as plants in the nursery industry. A TaqMan real-time PCR method for the detection of this pathogen in the field has been described previously; this paper describes the development of a number of assays based on this method which have various advantages for use in the field. A scorpion real-time PCR assay that is twice as fast as TaqMan was developed, allowing the detection of P. ramorum in less than 30 min. Also designed was a loop-mediated isothermal amplification (LAMP) assay, which allowed sensitive and specific detection of P. ramorum in 45 min using only a heated block. A positive reaction was identified by the detection of the LAMP product by color change visible to the naked eye.

show abstract

“…With Molecular beacon the pathogen was detected in a quantitative order similarly to TaqMan probe, which were able to detect levels as low as 1 fg DNA of the target pathogen present in plant tissues. In this study, the sensitivity of Molecular beacon and TaqMan probes against a dilution series of P. fragariae genomic DNA was equivalent (Bonants et al 2004). Also, Bilodeau et al (2007) compared three different chemistries including SYBR Green, TaqMan, and Molecular beacons using sequences of β-tubulin, ITS and elicitin gene regions for detection of P. ramorum causing sudden oak death.…”

Section: Detection Of Plant Pathogensmentioning

confidence: 96%

Real-time PCR applied to study on plant pathogens: potential applications in diagnosis - a review

Mirmajlessi¹,

Loit²,

Mänd³

et al. 2015

Plant Prot. Sci.

View full text Add to dashboard Cite

Mirmajlessi S.M., Loit E., Mänd M., Mansouripour S.M. (2015): Real-time PCR applied to study on plant pathogens: potential applications in diagnosis -a review. Plant Protect Sci., 51: 177-190.Quantitative real-time PCR (qPCR) technique incorporates traditional polymerase chain reaction (PCR) efficiency with the production of a specific fluorescent signal, measuring the kinetics of the reaction in the early PCR phases and providing quantification of specific targets in various environmental samples. There are an increasing number of chemistries to detect PCR products, which are widely used in plant pathology as they cluster into the amplicon sequence non-specific and sequence-specific techniques. In this review, we illustrate a general description of major chemistries and discuss some considerations for assay development as it applies for a wide range of applications in epidemiological studies. The technique has become the gold standard for early detection of pathogens and a fundamental tool in the research laboratory.

show abstract

A Combination of Baiting and Different PCR Formats, Including Measurement of Real-Time Quantitative Fluorescence, For the Detection of Phytophthora fragariae in Strawberry Plants

Cited by 41 publications

References 34 publications

A TaqMan PCR Method for Routine Diagnosis of the Quarantine Fungus Guignardia citricarpa on Citrus Fruit

A TaqMan PCR Method for Routine Diagnosis of the Quarantine Fungus Guignardia citricarpa on Citrus Fruit

Faster, Simpler, More-Specific Methods for Improved Molecular Detection of Phytophthora ramorum in the Field

Real-time PCR applied to study on plant pathogens: potential applications in diagnosis - a review

Contact Info

Product

Resources

About