Problems encountered during immunofluorescent diagnosis of fireblight

Roberts, Pauline

doi:10.1111/j.1365-3059.1980.tb01187.x

Cited by 23 publications

(10 citation statements)

References 2 publications

(1 reference statement)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Current approaches for the identification of fireblight use microbiological and molecular methods. Serological tests such as immunofluorescence and ELISA are time‐consuming and cause problems with sensitivity and specificity (Roberts, 1980). Polyclonal antibodies may cross‐react with other plant‐associated bacteria, resulting in false positive results.…”

Section: Introductionmentioning

confidence: 99%

Real‐time PCR for detection and quantification of Erwinia amylovora, the causal agent of fireblight

Salm

Geider

2004

Plant Pathology

View full text Add to dashboard Cite

Real-time PCR was used for quantitative detection of Erwinia amylovora , the causative agent of fireblight. Specific primers were created from a DNA fragment of the common plasmid pEA29, successfully used for standard PCR identification of the pathogen. The primers amplified DNA from various E. amylovora strains, but not from other plant-associated bacteria. DNA of E. amylovora was also amplified from field samples and from inoculated apple leaves or flowers. Neither the presence of other bacteria nor low amounts of tissue extracts from bark or leaves changed the signal threshold. Assays with SYBR Green I instead of the Taqman probe showed a similar sensitivity, detecting 50 cells per assay. Real-time PCR could be especially useful for mass screening of commercial products and for resistance studies of transgenic host plants, in breeding experiments and after treatments to control fireblight.

show abstract

Section: Introductionmentioning

confidence: 99%

Real‐time PCR for detection and quantification of Erwinia amylovora, the causal agent of fireblight

Salm

Geider

2004

Plant Pathology

View full text Add to dashboard Cite

show abstract

“…Long-distance as well as local spread of fire blight, caused by movement of E. amylovora-contaminated plant material, could be considerably minimized by monitoring such material for the pathogen by using sensitive detection techniques. Serological techniques (14), including the use of monoclonal antibodies (12) or semiselective media (13), can be applied in the case of massive contaminations, but latent infections with low concentrations of the pathogen (below 105 CFU/ml) require a more sensitive and reliable method to detect the bacterium in a field sample. Molecular biology has provided a powerful tool to identify E. amylovora.…”

Section: Discussionmentioning

confidence: 99%

“…However, plant material appears to be the most important source of fire blight epidemics, which may be confined by the monitoring of suspect plants lacking disease symptoms. Conventional diagnostic methods, including the use of semiselective media (13) or immunofluorescence techniques (14), cannot detect small quantities of E. amylovora, i.e., detection is correlated primarily with obvious disease symptoms. Therefore, we have developed DNA hybridization techniques with DNA from a 29-kb plasmid which is common to E. amylovora strains.…”

mentioning

confidence: 99%

Sensitive and species-specific detection of Erwinia amylovora by polymerase chain reaction analysis

Bereswill

Pahl

Bellemann

et al. 1992

Appl Environ Microbiol

188

View full text Add to dashboard Cite

Detection and identification of the fire blight pathogen, Erwinia amylovora, can be accurately done by polymerase chain reaction (PCR) analysis in less than 6 h. Two oligomers derived from a 29-kb plasmid which is common to all strains of E. amylovora were used to amplify a 0.9-kb fragment of the plasmid. By separation of the PCR products on agarose gel, this fragment was specifically detected when E. amylovora DNA was present in the amplification assay. It was not found when DNA from other plant-pathogenic bacteria was used for the assay. A visible band specific to the 0.9-kb fragment was produced with DNA from fewer than 100 E. amylovora cells. A signal of similar strength was also obtained from E. amylovora cell lysates in the presence of the mild detergent Tween 20. Signals were weaker when bacteria were added to the PCR mixture without the detergent. As with results obtained from hybridization experiments using pEA29 DNA, the PCR signal was obtained with E. amylovora isolates from various geographic regions. This technique could also be used for detection of the fire blight pathogen in extracts of tissue obtained from infected plant material.

show abstract

“…Specific direct observational methods are based on the use of antisera. For indirect immunofluorescence procedures (Roberts, 1980) a goat anti-rabbit fluorescein isocyanate-labelled conjugate is attached to rabbit serum which facilitates the recognition of bacterial cells by their yellow/green fluorescence under UV microscopy. Immunofluorescence has been used routinely for detection of fireblight on imported ornamentals (McCracken, 1988).…”

Section: Detection Of Bacterial Pathogens On Plant Surfacesmentioning

confidence: 99%

Pathogens in the foliar environment

Blakeman

1993

Plant Pathology

View full text Add to dashboard Cite

Problems encountered during immunofluorescent diagnosis of fireblight

Cited by 23 publications

References 2 publications

Real‐time PCR for detection and quantification of Erwinia amylovora, the causal agent of fireblight

Real‐time PCR for detection and quantification of Erwinia amylovora, the causal agent of fireblight

Sensitive and species-specific detection of Erwinia amylovora by polymerase chain reaction analysis

Pathogens in the foliar environment

Contact Info

Product

Resources

About