Rapid detection of potato viruses by dot-ELISA

Weidemann, H. L.

doi:10.1007/bf02357886

Cited by 16 publications

(4 citation statements)

References 16 publications

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“…For production of virus-free propagation material originating from in vitro cultures, detection of very low concentrations of virus particlesor viral RNA moleculesis required. The limit of virus detection by ELISA is within the range of 20-100 ng (Weidemann, 1988). The sensitivity of the dot-spot hybridisation method for the detection of plant viruses is in a range similar to that for the ELISA method (Rosner & Bar-Joseph, 1984).…”

Section: Introductionmentioning

confidence: 89%

The use of the polymerase chain reaction (PCR) for the detection of bean yellow mosaic virus in gladiolus

Vunsh¹,

Rosner²,

Stein³

1990

Annals of Applied Biology

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A method is described for the improved detection of bean yellow mosaic virus (BYMV) in gladioli leaves. Specific sequences of BYMV RNA, present in total RNA extracts of infected plants were detected following amplification by the polymerase chain reaction (PCR). The viral RNA was initially reverse-transcribed into cDNA, then specific sequences were amplified by PCR using specific oligonucleotides as primers. Detectable amounts of virus RNA in BYMV-infected plant tissue by PCR were approximately three to four orders of magnitude lower as compared with those detectable by ELISA and molecular hybridisation. Combining PCR with molecular hybridisation (using a 32P-labelled transcript of viral sequences as a probe), further increased the sensitivity of this method to a gain of four to five orders of magnitude as compared with direct molecular hybridisation, and enabled the detection of up to single picogram quantities of the virus.

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Section: Introductionmentioning

confidence: 89%

The use of the polymerase chain reaction (PCR) for the detection of bean yellow mosaic virus in gladiolus

Vunsh¹,

Rosner²,

Stein³

1990

Annals of Applied Biology

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“…The experiments confirmed that it is possible to evaluate a dot immunobinding assay for the detection of Phytophthora species with a polyclonal antiserum (Pab) in naturally dark-rooted older plants. Although in experiments with tomato roots and Phytophthora capsici (Kimishima & Kobayashi, 1990), cabbage roots and Plusmodiophoru brussicae (Lange et al, 1989) or other plant tissues and other pathogens (Bode, Beutin & Kohler, 1984;Hibi & Saito, 1985;Graddon & Randles, 1986;Powell, 1987;Smith & Banttari, 1987;Weidemann, 1988) non-specific reactions of the plant material could be successfully reduced with modifications of different steps; in the present study it was not possible to differentiate between infected and non-infected root samples by the occurrence or non-occurrence of a coloured spot. The ever present coloured spot, no matter whether the roots were healthy or not and no matter which modification of the different steps in the procedure were tested, suggest nonspecific background reactions of the ingredients in naturally dark roots.…”

Section: Discussionmentioning

confidence: 65%

Development of a dot immunobinding assay to detect Phytophthora spp. in naturally dark‐rooted woody plants

Hahn

Werres²

1997

Annals of Applied Biology

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An indirect dot immunobinding assay (DIBA) with a PAb raised against an isolate of P. cinnamomi was evaluated to detect Phytophthora spp. in naturally dark-rooted woody plants. Screening of different modifications of the procedure were done with root samples of Chamaecyparis lawsoniana non-infected and infected with P. cinnamomi. With the optimised DIBA procedure, root infection could be diagnosed by a clearly defined halo around the spot and by a colour change in the spot using Fast Red or Fast Blue substrate. Detection of different Phytophthora species in Chamaecyparis plants from commercial nurseries was successful with the optimised procedure.

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“…The time taken per assay can be reduced by incubating enzyme conjugated virus-specific antibodies and sample together in the wells of the plate (Van Vuurde and Maat, 1985;Van den Heuvel and Peters, 1989). Squash or dot blot tests can be done on individual leaf samples (taken in the field) by pressing leaf sap onto a support membrane (nitrocellulose, nylon or even notepaper) (Heide and Lange, 1988;Weidemann, 1988;Mitchell et al, 1990).…”

Section: Potato Virus Testingmentioning

confidence: 99%

Developments in methodology of plant virus detection

Torrance¹

1992

Netherlands Journal of Plant Pathology

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As a general rule some form of polyclonal-antibody based enzyme immunoassay is still preferred for routine plant virus detection, but modifications may be necessary when increased sensitivity or specificity is required. In recent years new developments in antibody-based detection methods have mostly involved the provision of specific reagents, such as monoclonal antibodies or affinity-purified second antibody-enzyme conjugates which have helped to improve the sensitivity and specificity of the assays. Other kinds of tests (such as nucleic acid hybridisation) must be used when tests for virus coat protein are not appropriate. Recently, tests based on the polymerase chain reaction show great promise. There is no single universal test: assays must be devised and optimised for each pathogen. The challenge with all types of test is to make them quicker, less labour intensive, andif possible -cheaper.

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Rapid detection of potato viruses by dot-ELISA

Cited by 16 publications

References 16 publications

The use of the polymerase chain reaction (PCR) for the detection of bean yellow mosaic virus in gladiolus

The use of the polymerase chain reaction (PCR) for the detection of bean yellow mosaic virus in gladiolus

Development of a dot immunobinding assay to detect Phytophthora spp. in naturally dark‐rooted woody plants

Developments in methodology of plant virus detection

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