A combination of baiting and different PCR formats, including measurement of real-time quantitative fluorescence, for the detection of Phytophthora fragariae in strawberry plants

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

    Research output: Contribution to journalArticleAcademicpeer-review

    30 Citations (Scopus)

    Abstract

    Phytophthora fragariae, the cause of strawberry red stele disease, is a quarantine pathogen in Europe. Detecting low levels of infection requires sensitive and specific methods. In the past, Dutch and English inspection services have used bait plants to test strawberry propagation stocks destined for export. Increasingly though, PCR is being incorporated into these testing procedures in an effort to increase sensitivity and speed. Various combinations of baiting and PCR assays were compared with existing testing procedures. Water and root samples from the bait test were screened by nested PCR and the PCR amplicon was detected by several methods, including fluorescent labelled probes (TaqMan and Molecular Beacon). PCR amplification was monitored in real-time and semi-quantitative detection was possible. Because PCR reactions are sensitive to inhibitors present in extracted DNA samples, an internal control containing the primer sequences specific for P. fragariae was developed to avoid false negatives
    Original languageEnglish
    Pages (from-to)689-702
    JournalEuropean Journal of Plant Pathology
    Volume110
    Issue number7
    DOIs
    Publication statusPublished - 2004

    Fingerprint

    Phytophthora fragariae
    baiting
    strawberries
    fluorescence
    baits
    testing
    stele
    quarantine
    sampling
    pathogens
    DNA
    assays
    methodology
    infection
    water

    Keywords

    • polymerase-chain-reaction
    • molecular beacons
    • dna
    • hybridization
    • infestans
    • identification
    • amplification
    • rna
    • quantification
    • organization

    Cite this

    @article{c3ad59e4d2da42b2b5482fd82958cd01,
    title = "A combination of baiting and different PCR formats, including measurement of real-time quantitative fluorescence, for the detection of Phytophthora fragariae in strawberry plants",
    abstract = "Phytophthora fragariae, the cause of strawberry red stele disease, is a quarantine pathogen in Europe. Detecting low levels of infection requires sensitive and specific methods. In the past, Dutch and English inspection services have used bait plants to test strawberry propagation stocks destined for export. Increasingly though, PCR is being incorporated into these testing procedures in an effort to increase sensitivity and speed. Various combinations of baiting and PCR assays were compared with existing testing procedures. Water and root samples from the bait test were screened by nested PCR and the PCR amplicon was detected by several methods, including fluorescent labelled probes (TaqMan and Molecular Beacon). PCR amplification was monitored in real-time and semi-quantitative detection was possible. Because PCR reactions are sensitive to inhibitors present in extracted DNA samples, an internal control containing the primer sequences specific for P. fragariae was developed to avoid false negatives",
    keywords = "polymerase-chain-reaction, molecular beacons, dna, hybridization, infestans, identification, amplification, rna, quantification, organization",
    author = "P.J.M. Bonants and {van Gent-Pelzer}, M.P.E. and R. Hooftman and D.E.L. Cooke and D.C. Guy and J.M. Duncan",
    year = "2004",
    doi = "10.1023/B:EJPP.0000041551.26970.0e",
    language = "English",
    volume = "110",
    pages = "689--702",
    journal = "European Journal of Plant Pathology",
    issn = "0929-1873",
    publisher = "Springer Verlag",
    number = "7",

    }

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

    In: European Journal of Plant Pathology, Vol. 110, No. 7, 2004, p. 689-702.

    Research output: Contribution to journalArticleAcademicpeer-review

    TY - JOUR

    T1 - A combination of baiting and different PCR formats, including measurement of real-time quantitative fluorescence, for the detection of Phytophthora fragariae in strawberry plants

    AU - Bonants, P.J.M.

    AU - van Gent-Pelzer, M.P.E.

    AU - Hooftman, R.

    AU - Cooke, D.E.L.

    AU - Guy, D.C.

    AU - Duncan, J.M.

    PY - 2004

    Y1 - 2004

    N2 - Phytophthora fragariae, the cause of strawberry red stele disease, is a quarantine pathogen in Europe. Detecting low levels of infection requires sensitive and specific methods. In the past, Dutch and English inspection services have used bait plants to test strawberry propagation stocks destined for export. Increasingly though, PCR is being incorporated into these testing procedures in an effort to increase sensitivity and speed. Various combinations of baiting and PCR assays were compared with existing testing procedures. Water and root samples from the bait test were screened by nested PCR and the PCR amplicon was detected by several methods, including fluorescent labelled probes (TaqMan and Molecular Beacon). PCR amplification was monitored in real-time and semi-quantitative detection was possible. Because PCR reactions are sensitive to inhibitors present in extracted DNA samples, an internal control containing the primer sequences specific for P. fragariae was developed to avoid false negatives

    AB - Phytophthora fragariae, the cause of strawberry red stele disease, is a quarantine pathogen in Europe. Detecting low levels of infection requires sensitive and specific methods. In the past, Dutch and English inspection services have used bait plants to test strawberry propagation stocks destined for export. Increasingly though, PCR is being incorporated into these testing procedures in an effort to increase sensitivity and speed. Various combinations of baiting and PCR assays were compared with existing testing procedures. Water and root samples from the bait test were screened by nested PCR and the PCR amplicon was detected by several methods, including fluorescent labelled probes (TaqMan and Molecular Beacon). PCR amplification was monitored in real-time and semi-quantitative detection was possible. Because PCR reactions are sensitive to inhibitors present in extracted DNA samples, an internal control containing the primer sequences specific for P. fragariae was developed to avoid false negatives

    KW - polymerase-chain-reaction

    KW - molecular beacons

    KW - dna

    KW - hybridization

    KW - infestans

    KW - identification

    KW - amplification

    KW - rna

    KW - quantification

    KW - organization

    U2 - 10.1023/B:EJPP.0000041551.26970.0e

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    M3 - Article

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    SN - 0929-1873

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