Application of an Enzyme-Linked Immunosorbent Assay (ELISA) involving Monoclonal Antibody for Detection of BLV Antibodies in Individual or Pooled Bovine Milk Samples

2010 ◽  
Vol 32 (1-10) ◽  
pp. 526-533 ◽  
Author(s):  
M. Mammerickx ◽  
D. Portetelle ◽  
A. Burny
1990 ◽  
Vol 53 (7) ◽  
pp. 577-580 ◽  
Author(s):  
JUAN I. AZCONA ◽  
MOHAMED M. ABOUZIED ◽  
JAMES J. PESTKA

A hybridoma-based method utilizing tandem affinity chromatography and enzyme-linked immunosorbent assay (ELISA) was devised to detect zearalenone. A zearalenone specific monoclonal antibody was attached to Sepharose for initial sample clean-up. Zearalenone was eluted with methanol and then quantified by competitive direct ELISA. Average ELISA recoveries from the column for water spiked with zearalenone at levels of 1, 5, 10, 25, and 50 ng/ml were 107, 86, 95, 95, and 92%, respectively, with a mean recovery of 95%. Mean interwell and interassay coefficients of variation were 9.7 and 8.9%, respectively. Average recovery by the method from milk spiked with zearalenone at levels of 1, 5, 10, 25, and 50 ng/ml was 187, 113, 107, 110, and 112%, respectively, with a mean recovery of 126%. Mean interwell and interassay coefficients of variation were 14.5 and 9.1%, respectively. Zearalenone was not detectable in 12 commercial milk samples assayed by the tandem method.


1997 ◽  
Vol 60 (1) ◽  
pp. 64-66 ◽  
Author(s):  
G. ANGUITA ◽  
R. MARTÍN ◽  
T. GARCÍA ◽  
P. MORALES ◽  
A. I. HAZA ◽  
...  

A competitive ELISA (enzyme-linked immunosorbent assay) was performed to detect and quantify bovine milk in ovine and caprine milk and cheese using a monoclonal antibody (AH4 MAb) against bovine β-casein. Ovine or caprine milk and cheese containing bovine milk were added simultaneously with the AH4 MAb to the wells of a microtiter plate that had been previously sensitized with commercial bovine β-casein. The bovine caseins in milk or cheese samples compete with the bovine β-casein bound to the plate for the AH4 MAb binding sites. Further immunorecognition of AH4 MAb bound to the bovine β-casein immobilized onto the plate was attained with rabbit anti-mouse immunoglobulin conjugated to peroxidase. Subsequent enzymic conversion of the substrate showed clear differences in absorbance values during assay of mixtures of ovine and caprine milk and cheese containing various amounts of bovine milk. The competitive ELISA developed in this work allows the quantitative detection of bovine milk in ovine and caprine milk and cheese samples in the range of 0.5 to 25% of substitution.


1992 ◽  
Vol 59 (2) ◽  
pp. 123-133 ◽  
Author(s):  
Catherine A. O'Sullivan ◽  
Patrick J. Joyce ◽  
Teresa Sloan ◽  
Alan G. Shattock

SummaryA direct capture enzyme-linked immunosorbent assay (ELISA) was developed to measure elevated polymorphonuclear granulocyte (PMN) antigens using horseradish peroxidase (EC 1.11.1.7) conjugated rabbit polyclonal anti-PMN antisera and a monoclonal antibody specific for PMN cells. Optical densities obtained in the ELISA were used to predict the cell counts of milk samples. Predicted counts were not significantly different from actual somatic cell counts (SCC). In a total of 156 bovine milk samples the correlation coefficient between somatic cell counting, taking > 500000 cells/ml as being indicative of mastitis, and the assay was 0·94, yielding an assay sensitivity of 95·2% and a specificity of 97·3%. In further trials the ELISA could detect elevated PMN antigens in milk with SCC as low as 100000 cells/ml. The results indicate that the monoclonal antibody based direct ELISA has excellent potential in the detection and determination of bovine mastitis.


2006 ◽  
Vol 13 (3) ◽  
pp. 420-422 ◽  
Author(s):  
S. E. Burastero ◽  
C. Paolucci ◽  
D. Breda ◽  
G. Monasterolo ◽  
R. E. Rossi ◽  
...  

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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