An enzyme-linked immunosorbent assay for the detection of Rathayibacter toxicus, the bacterium involved in annual ryegrass toxicity, in hay

2006 ◽  
Vol 57 (7) ◽  
pp. 731 ◽  
Author(s):  
A. M. Masters ◽  
A. R. Gregory ◽  
R. J. Evans ◽  
J. E. Speijers ◽  
S. S. Sutherland
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Specific Antigen ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Annual Ryegrass ◽  
Capture Assay ◽  
Large Numbers ◽  
Soluble Polysaccharide ◽  
Annual Ryegrass Toxicity

An enzyme-linked immunosorbent assay (ELISA) for Rathayibacter toxicus is described. The development of a monoclonal antibody for a specific antigen from R. toxicus and a polyclonal antibody raised against the same R. toxicus preparation enabled a capture assay format. The assay is specific for a soluble polysaccharide produced by the bacterium and was found to be sensitive enough to detect antigen equivalent to less than one gall per kilogram of hay. The applicability of the assay to samples of pasture or hay is demonstrated. Cost-effective testing of large numbers of samples for the presence of R. toxicus is possible with the ELISA. This will assist stockowners, hay producers, and hay exporters in the management of the risk of annual ryegrass toxicity.

Improvements to the immunoassay for detection of Rathayibacter toxicus in hay

2011 ◽  
Vol 62 (6) ◽  
pp. 523 ◽  
Author(s):  
A. M. Masters ◽  
B. Samarasinghe ◽  
M. J. Kalkhoven ◽  
G. L. den Hollander ◽  
D. G. Palmer
Keyword(s):  
Monoclonal Antibody ◽  
Horseradish Peroxidase ◽  
Immunosorbent Assay ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Export Industry ◽  
Large Numbers ◽  
Assay Protocol ◽  
Very High ◽  
Peroxidase Conjugate

An improved protocol for the previously described enzyme-linked immunosorbent assay for Rathayibacter toxicus in hay is described. The improvements were driven mainly by the export hay industry requirement of same-day turnaround for testing of hay extracts. The preparation of hay extracts was shortened by 8 h. The time for adding samples to the enzyme-linked immunosorbent assay plates was shortened by the use of sample tubes with penetrable stoppers combined with specially designed racks. The monoclonal antibody used in the original protocol was purified and conjugated to horseradish peroxidase. This eliminated the need for a secondary step with an anti-mouse horseradish peroxidase conjugate and thereby shortened the assay by over 1 h. Results with the improved assay protocol showed a very high correlation with results obtained with the original protocol (r = 0.98). The assay is still sensitive enough to detect antigen equivalent to less than 1 average gall per kg of hay. These cost-effective changes have streamlined the testing of large numbers of samples for the presence of R. toxicus, in support of the hay export industry.

Development and application of polymerase chain reaction-based assays for Rathayibacter toxicus and a bacteriophage associated with annual ryegrass (Lolium rigidum) toxicity

2007 ◽  
Vol 47 (2) ◽  
pp. 177 ◽  
Author(s):  
M. C. Kowalski ◽  
D. Cahill ◽  
T. J. Doran ◽  
S. M. Colegate
Keyword(s):  
Polymerase Chain Reaction ◽  
Immunosorbent Assay ◽  
Potential Application ◽  
South Australia ◽  
Enzyme Linked Immunosorbent Assay ◽  
Annual Ryegrass ◽  
Lolium Rigidum ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Annual Ryegrass Toxicity

Annual ryegrass toxicity (ARGT) is responsible for significant stock losses in South Australia and Western Australia. The toxicity is caused by corynetoxins produced by the bacterium Rathayibacter toxicus (with the possible involvement of a bacteriophage), which infects annual ryegrass (Lolium rigidum). Polymerase chain reaction (PCR)-based assays, compatible with an existing enzyme-linked immunosorbent assay for the corynetoxins, have been developed and used to screen L. rigidum for both the presence of R. toxicus and for the bacteriophage isolate NCPPB 3778. The results from analysing bacterially infected galls from toxic grain screenings showed a positive correlation between the presence of the bacterium and corynetoxins but not with the bacteriophage. Analysis of pasture-derived samples of annual ryegrass showed about a 50% correlation of corynetoxins with bacterial presence and about a 5% correlation of phage with the presence of the bacterium. These observations support the potential application of the PCR-based assays in providing a useful, complementary tool in the assessment of the likelihood of pasture and feed to cause ARGT and to enable a better understanding of the complex aetiology of ARGT.

A semi-quantitative enzyme-linked immunosorbent assay for Rathayibacter toxicus, the bacterium involved in annual ryegrass toxicity, to assist in risk assessment of fodder for domestic use

2014 ◽  
Vol 65 (12) ◽  
pp. 1329 ◽  
Author(s):  
A. M. Masters ◽  
B. Samarasinghe ◽  
M. Kalkhoven ◽  
L. den Hollander ◽  
D. G. Palmer
Keyword(s):  
Immunosorbent Assay ◽  
Animal Health ◽  
Clinical Signs ◽  
Risk Level ◽  
Enzyme Linked Immunosorbent Assay ◽  
Annual Ryegrass ◽  
Department Of Agriculture ◽  
Domestic Livestock ◽  
Annual Ryegrass Toxicity ◽  
Risk Categories

A semi-quantitative enzyme-linked immunosorbent assay (ELISA) for the detection of Rathayibacter toxicus in hay or pasture was used to estimate the degree of R. toxicus bacterial gall contamination in hay or pasture that was unsuitable for export but that may be suitable for feeding to domestic livestock. Based on experience of testing of fodder samples from pastures where livestock showed clinical signs, or outbreaks of annual ryegrass toxicity (ARGT), four relevant levels of bacterial gall contamination were selected. Several 1-kg samples of hay with no contamination by R. toxicus were spiked with these four different amounts of bacterial galls to provide five different risk categories considered relevant to the occurrence of ARGT. Extracts of the spiked samples were assayed to determine the range of ELISA results to be expected in each category. To validate these risk categories, all cases of ARGT diagnosed between 2000 and 2012 by the Animal Health Laboratories, Department of Agriculture and Food Western Australia, associated with the submission of suspected toxic fodder were allocated to the risk categories on the basis of the recorded results for fodder. In ~15% and 79% of all cases, the fodder associated with outbreaks of ARGT fell into the ‘moderate risk’ and ‘high risk’ categories, respectively. The selected categories were considered to provide realistic estimations of the risk that fodder within them might cause ARGT if fed to livestock. This risk-level reporting has been adopted to enable informed decision making as to whether feeding a particular batch of hay or fodder to animals represents an acceptable risk.

scholarly journals Preparation of Monoclonal Antibody and Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay and Fluorescence-Linked Immunosorbent Assay for Detecting 3-Amino-5-methylmorpholino-2-oxazolidinone (AMOZ) in Edible Animal Tissue

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4243
Author(s):  
Yong Xie ◽  
Yarong Wang ◽  
Xueling Yan ◽  
Lu Gan ◽  
Tao Le
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Correlation Coefficients ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Monitoring Methods ◽  
Animal Tissues ◽  
Coefficients Of Variation ◽  
Highly Sensitive ◽  
Ic50 Values

To monitor the illegal used of furaltadone, a highly sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and fluorescence-linked immunosorbent assay (FLISA) based on a monoclonal antibody (mAb) were developed for the detection of 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ), the major metabolite of furaltadone in animal tissues. The highly specific mAb, which was very sensitive to a nitrophenyl derivative of AMOZ (2-NP-AMOZ) with IC50 values of 0.11 and 0.09 ng/mL for ic-ELISA and FLISA, respectively, was selected for the development of immunoassays. For both the ic-ELISA and FLISA for AMOZ-spiked experiments, acceptable recovery rates of 81.1–105.3% and coefficients of variation of 4.7–9.8% were obtained. In addition, results from both ic-ELISA and FLISA methods for spiked samples’ data showed excellent correlation coefficients ranging from 0.9652 to 0.9927. Meanwhile, the proposed ic-ELISA and FLISA for thirty spiked samples were confirmed by standard LC-MS/MS with high correlation coefficients of 0.9911 and 0.9921, respectively. These results suggest that the developed ic-ELISA and FLISA are valid and cost-effective tools for high-throughput monitoring methods for AMOZ residues in animal tissues.

Evaluation of a human pancreas-specific antigen by enzyme-linked immunosorbent assay

1981 ◽  
Vol 117 (3) ◽  
pp. 251-258 ◽  
Author(s):  
Rueyming Loor ◽  
Manabu Kuriyama ◽  
Mary Lou Manzo ◽  
Hideo Inaji ◽  
Harold O. Douglass ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Specific Antigen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Pancreas

scholarly journals Unreliable Measurement of Basophil Maximum Leukotriene Release with the Bühlmann CAST 2000 Enzyme-Linked Immunosorbent Assay Kit

2006 ◽  
Vol 13 (3) ◽  
pp. 420-422 ◽  
Author(s):  
S. E. Burastero ◽  
C. Paolucci ◽  
D. Breda ◽  
G. Monasterolo ◽  
R. E. Rossi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
False Positive ◽  
Positive Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Positive Results

ABSTRACT The Bühlmann CAST 2000 enzyme-linked immunosorbent assay is a potentially useful assay for measuring sulfidoleukotrienes released in vitro by allergen-challenged basophils. However, we observed that the positive-control reagent yielded positive signals in cell-free systems. These false-positive results depended on using a mouse anti-FcεRI monoclonal antibody and were prevented by degranulation-inducing reagents other than mouse monoclonal antibodies.

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