Immunological techniques used with fungal plant pathogens: aspects of antigens, antibodies and assays for diagnosis

Werres, Sabine; Steffens, Claudia

doi:10.1111/j.1744-7348.1994.tb04998.x

Cited by 32 publications

(24 citation statements)

References 135 publications

(85 reference statements)

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“…Antibody based technology is frequently employed in plant pathogen immunodiagnostic systems (WERRES & STEFFENS 1994;DEWEY & THORNTON 1995). Using these techniques to detect and quantify target airborne spora rapidly will prove useful both as an immediate measure of spatial distribution of plant pathogenic fungi and as a research tool in quantatitive epidemiology.…”

Section: Discussionmentioning

confidence: 99%

New methods for detecting and enumerating fungal spores of plant pathogens

Kennedy¹,

Wakeham²

2002

Plant Prot. Sci.

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Information on the presence or absence of airborne spores or other particles would be useful in an increasing number of areas including agriculture. Traditional methods used for detecting and enumerating of airborne spores of fungal plant pathogens are time consuming and require specialist knowledge. Some spore types (e.g. ascospores) are difficult to differentiate using these methods. To facilitate this, new methods, which can be used to accurately differentiate fungal spore types, are required. A Burkard 7-day volumetric spore trap used in combination with an immunofluorescence test has been developed to detect and quantify field-trapped ascosporic inoculum of Mycosphaerella brassicicola (the ringspot pathogen of brassicas). This test has also been found useful in the validation of more rapid user-friendly immunoassay based trapping procedures. A microtiter immunospore trapping device, which uses a suction system to directly trap airparticulates by impaction into microtiter wells, has been used successfully for the rapid detection and quantification of ascosporic inoculum of M. brassicicola. The system shows potential for the rapid field-detection of airborne ascosporic inoculum of the ringspot pathogen.

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Section: Discussionmentioning

confidence: 99%

New methods for detecting and enumerating fungal spores of plant pathogens

Kennedy¹,

Wakeham²

2002

Plant Prot. Sci.

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“…With the wide range of fungal species tested, it was interesting to note such a low level of crossreactivity. Problems of specificity with polyclonal antibodies in fungal diagnostic assays have been extensively documented (Werres & Steffens, 1994;Dewey & Thornton, 1995). However, the cross-reactivity noted to the conidial wall of P. brassicae and M. pinodes, and to the ascospore wall of a number of fungal spp.…”

Section: R Kennedy Et Al 304mentioning

confidence: 99%

“…However, such methods require considerable amounts of time and expertise if accurate counts are to be obtained and are not practical or accurate where several related species of fungi that have morphologically similar spore types (such as ascosporic fungi) occur together. Technological advances in fungal diagnostics, using either antibody or nucleic acid probes (Werres & Steffens, 1994;Beck et al, 1996), may be appropriate for the specific and quantitative detection of airborne ascospores of M. brassicicola. Immunoassays are increasingly exploited because of their speed, relatively low cost, simplicity and the ability to perform on-site quantitative assays.…”

Section: Introductionmentioning

confidence: 99%

Production and immunodetection of ascospores of Mycosphaerella brassicicola: ringspot of vegetable crucifers

Kennedy¹,

Wakeham²,

Cullington³

1999

Plant Pathology

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Pseudothecia containing abundant ascospores of Mycosphaerella brassicicola were produced in vitro on Brussels sprout decoction agar at 15ЊC under a 16-hour photoperiod of different light regimes. Spermogonia containing spermatia were also produced on the decoction agar. Ascospores were released when cultures were misted with SDW and placed under continuous light. Germination of ascospores was highest between 20ЊC and 25ЊC and spores remained viable at relative humidities above 93·5%. Exposure of ascospores to 55% relative humidity for 24 h reduced their germination to 75%. A polyclonal antiserum raised against whole ascospores was used to detect, by immunofluorescence, the ascospore and mycelial wall of M. brassicicola, following reaction with anti-rabbit IgG FITC conjugate. Autofluorescence of spore and mycelial components of other fungal species could be eliminated using the counterstains Evan's blue and eriochrome black at 0·2% and 0·5%, respectively, in phosphate buffered saline (pH 7·2). A procedure was developed to detect, by immunofluorescence, ascospores of M. brassicicola on artificially inoculated Melinex spore tape. Coating of the spore tape with bovine serum albumin provided a suitable support medium and blocking agent for detection of ascospores in the field. The potential use of the system for selective detection of ascospores of M. brassicicola in infected crops of vegetable brassicas in the presence of other ascosporic fungi is discussed.

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“…The generic biosensor setup described in this paper could feasibly be used for detection of any fungal spore, the only requirement being the availability of a suitable antibody specifically recognizing only corresponding target antigens. Various specific antibodies, including polyclonals, monoclonals and single-chain Fv antibody fragments (scFv), have been developed against multiple important fungal and fungal-like spore species [26][27][28]. By immobilising a suitable capture molecule on the sensor chip, such as an anti-isotype antibody, antiFab antibody or protein A/G affinity purified antibody, all available as pathogen specific antibodies can potentially be used for label-free detection using the approach described in this paper.…”

Section: Cross-reactivity Studiesmentioning

confidence: 99%

Development of Surface Plasmon Resonance (SPR) Based Immuno-Sensing System for Detection of Fungal Teliospores of Karnal Bunt (Tilletia indica), a Quarantined Disease of Wheat

Singh¹

2012

J Biosens Bioelectron

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Immunological techniques used with fungal plant pathogens: aspects of antigens, antibodies and assays for diagnosis

Cited by 32 publications

References 135 publications

New methods for detecting and enumerating fungal spores of plant pathogens

New methods for detecting and enumerating fungal spores of plant pathogens

Production and immunodetection of ascospores of Mycosphaerella brassicicola: ringspot of vegetable crucifers

Development of Surface Plasmon Resonance (SPR) Based Immuno-Sensing System for Detection of Fungal Teliospores of Karnal Bunt (Tilletia indica), a Quarantined Disease of Wheat

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