Immunostick Colorimetric Assay for Detection of Pseudomonas spp. in Refrigerated Meat and Milk

1997 ◽  
Vol 60 (8) ◽  
pp. 908-911 ◽  
Author(s):  
ROSALBA GUTIÉRREZ ◽  
TERESA GARCÍA ◽  
ISABEL GONZÁLEZ ◽  
BERNABÉ SANZ ◽  
PABLO E. HERNÁNDEZ ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Pseudomonas Fluorescens ◽  
Colorimetric Assay ◽  
Detection Threshold ◽  
Live Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Pseudomonas Spp ◽  
Milk Samples ◽  
Elisa Format

Monoclonal antibodies generated against live cells of Pseudomonas fluorescens have been used in an immunostick enzyme-linked immunosorbent assay (ELISA) format for the detection of Pseudomonas spp. in refrigerated meat and milk. The detection threshold for the immunostick ELISA assay developed in this work is 104 CPU cm−2 for meat and 105 CPU ml−1 for milk samples.

Monoclonal Antibodies and an Indirect ELISA for Detection of Psychrotrophic Bacteria in Refrigerated Milk

1997 ◽  
Vol 60 (1) ◽  
pp. 23-27 ◽  
Author(s):  
ROSALBA GUTIÉRREZ ◽  
ISABEL GONZÁLEZ ◽  
TERESA GARCÍA ◽  
ESTER CARRERA ◽  
BERNABÉ SANZ ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Horseradish Peroxidase ◽  
Indirect Elisa ◽  
Detection Threshold ◽  
Microtiter Plate ◽  
Live Cells ◽  
Elisa Assay ◽  
Psychrotrophic Bacteria ◽  
Milk Samples ◽  
Elisa Format

Monoclonal antibodies generated against live cells of Pseudomonas fluorescens have been used in an indirect ELISA format for the detection of Pseudomonas spp. and related psychrotrophic bacteria in refrigerated milk. The immunorecognition of monoclonal antibodies adsorbed to bacteria bound to the wells of a microtiter plate was performed with rabbit anti-mouse immunoglobulins conjugated to horseradish peroxidase. Subsequent enzymic conversion of the substrate resulted in distinct absorbance differences when assaying milk samples containing psychrotrophic bacteria in the range 105 to 109 CFU ml−1. The detection threshold for the ELISA assay developed in this work is 105 CFU ml−1.

Demonstrating the Use of Bisphenol A-functionalised Gold Nanoparticles in Immunoassays

2013 ◽  
Vol 66 (6) ◽  
pp. 613 ◽  
Author(s):  
Joshua R. Peterson ◽  
Yang Lu ◽  
Erwann Luais ◽  
Nanju Alice Lee ◽  
J. Justin Gooding
Keyword(s):  
Gold Nanoparticles ◽  
Bisphenol A ◽  
Organic Molecule ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Aqueous Environments ◽  
Elisa Format ◽  
Hydrophobic Nature ◽  
Spherical Gold Nanoparticles ◽  
Antibody Interactions

Spherical gold nanoparticles (5-nm diameter) were modified with a small-molecule thiolated bisphenol A (BPA) ligand to achieve an estimated coverage of ~3.3 × 10–10 mol cm–2, or 180 ligands per particle. The modified particles were tested in an enzyme-linked immunosorbent assay (ELISA) format to measure functionality and were shown to bind specifically to anti-BPA antibody while resisting the non-specific adsorption of an antibody with no affinity for BPA. It was found that the use of 10 % ethanol as a co-solvent was required in the ELISA as aqueous buffers alone resulted in poor binding between anti-BPA antibody and the functionalised nanoparticles. This is likely due to the hydrophobic nature of the BPA ligand limiting its solubility, and therefore its availability for antibody interactions, in purely aqueous environments. To our knowledge, this is the first example of a nanoparticle modified with a small organic molecule being used in an ELISA assay.

scholarly journals Development and Application of a Novel Rapid and Throughput Method for Broad-Spectrum Anti-Foodborne Norovirus Antibody Testing

2021 ◽  
Vol 12 ◽  
Author(s):  
Yueting Zuo ◽  
Liang Xue ◽  
Junshan Gao ◽  
Yingyin Liao ◽  
Yueting Jiang ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Broad Spectrum ◽  
High Throughput Screening ◽  
Indirect Elisa ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Antibody Testing ◽  
Coefficients Of Variation ◽  
P Proteins

Foodbone norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Candidate vaccines are being developed, however, no licensed vaccines are currently available for managing NoV infections. Screening for stimulated antibodies with broad-spectrum binding activities can be performed for the development of NoV polyvalent vaccines. In this study, we aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA) for testing the broad spectrum of anti-NoV antibodies. Capsid P proteins from 28 representative NoV strains (GI.1–GI.9 and GII.1–GII.22 except GII.11, GII.18, and GII.19) were selected, prepared, and used as coating antigens on one microplate. Combined with incubation and the horseradish peroxidase chromogenic reaction, the entire process for testing the spectrum of unknown antibodies required 2 h for completion. The intra-assay and inter-assay coefficients of variation were less than 10%. The new method was successfully performed with monoclonal antibodies and polyclonal antibodies induced by multiple antigens. In conclusion, the indirect ELISA assay developed in this study had a good performance of reliability, convenience, and high-throughput screening for broad-spectrum antibodies.

Detection of Pseudomonas fluorescens and Related Psychrotrophic Bacteria in Refrigerated Meat by a Sandwich ELISA

1994 ◽  
Vol 57 (8) ◽  
pp. 710-714 ◽  
Author(s):  
I. GONZÁLEZ ◽  
R. MARTÍN ◽  
T. GARCÍA ◽  
P. MORALES ◽  
B. SANZ ◽  
...  
Keyword(s):  
Pseudomonas Fluorescens ◽  
Polyclonal Antibodies ◽  
Cell Envelope ◽  
Sandwich Elisa ◽  
Detection Threshold ◽  
Enzyme Linked Immunosorbent Assay ◽  
Psychrotrophic Bacteria ◽  
Specific Antigens ◽  
Protein F ◽  
Meat Samples

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for detection of Pseudomonas fluorescens and related psychrotrophic bacteria in refrigerated meat. Polyclonal antibodies were raised in rabbits against protein F from the cell envelope of P. fluorescens AH-70. The ELISA involved capturing antigens from the microorganisms present on meat samples with the anti-protein F antibodies immovilized on 96-well plates, and detecting bound antigens with the same anti-PF antibodies conjugated to biotin. Commercial ExtrAvidin-peroxidase conjugate was used to detect the biotinylated antibodies bound to their specific antigens. Subsequent enzymatic conversion of substrate gave distinct absorbance differences when assaying meat samples containing P. fluorescens strains of different origin as well as related psychrotrophic microorganisms. The detection threshold for the ELISA assay developed in this work is 105 CFU/cm2.

D-Dimer Determination to Exclude Pulmonary Embolism: a Two-Step Approach Using Latex Assay as a Screening Tool

1994 ◽  
Vol 72 (01) ◽  
pp. 089-091 ◽  
Author(s):  
P de Moerloose ◽  
Ph Minazio ◽  
G Reber ◽  
A Perrier ◽  
H Bounameaux
Keyword(s):  
Pulmonary Embolism ◽  
Screening Tool ◽  
Screening Tests ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Elisa Technique ◽  
Diagnostic Step ◽  
D Dimer ◽  
Rule Out ◽  
Latex Test

SummaryD-dimer (DD), when measured by a quantitative enzyme-linked immunosorbent assay (ELISA), is a valuable test to exclude venous thromboembolism (VTE). However, DD ELISA technique is not appropriate for emergency use and the available agglutination latex assays are not sensitive enough to be used as an alternative to rule out the diagnosis of VTE. Latex assays could still be used as screening tests. We tested this hypothesis by comparing DD levels measured by ELISA and latex assays in 334 patients suspected of pulmonary embolism. All but one patient with a positive (DD ≥500 ng/ml) latex assay had DD levels higher than 500 ng/ml with the ELISA assay. Accordingly, ELISA technique could be restricted to patients with a negative result in latex assay. This two-step approach would have spared about 50% of ELISA in our cohort. In conclusion, our data indicate that a latex test can be used as a first diagnostic step to rule out pulmonary embolism provided a negative result is confirmed by ELISA and the performance of the latex assay used has been assessed properly.

scholarly journals Epitope Mapping of Monoclonal Antibodies to Calreticulin Reveals That Charged Amino Acids Are Essential for Antibody Binding

Antibodies ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 31
Author(s):  
Ann Christina Bergmann ◽  
Cecilie Kyllesbech ◽  
Rimantas Slibinskas ◽  
Evaldas Ciplys ◽  
Peter Højrup ◽  
...  
Keyword(s):  
Amino Acids ◽  
Monoclonal Antibodies ◽  
Epitope Mapping ◽  
Biological Function ◽  
Specific Binding ◽  
Enzyme Linked Immunosorbent Assay ◽  
Potential Therapeutic Target ◽  
Charged Amino Acids ◽  
Chaperone Protein ◽  
Terminal Amino

Calreticulin is a chaperone protein, which is associated with myeloproliferative diseases. In this study, we used resin-bound peptides to characterize two monoclonal antibodies (mAbs) directed to calreticulin, mAb FMC 75 and mAb 16, which both have significantly contributed to understanding the biological function of calreticulin. The antigenicity of the resin-bound peptides was determined by modified enzyme-linked immunosorbent assay. Specific binding was determined to an 8-mer epitope located in the N-terminal (amino acids 34–41) and to a 12-mer peptide located in the C-terminal (amino acids 362–373). Using truncated peptides, the epitopes were identified as TSRWIESK and DEEQRLKEEED for mAb FMC 75 and mAb 16, respectively, where, especially the charged amino acids, were found to have a central role for a stable binding. Further studies indicated that the epitope of mAb FMC 75 is assessable in the oligomeric structure of calreticulin, making this epitope a potential therapeutic target.

scholarly journals A Survey of G6 and G10 Serotypes of Group a Bovine Rotaviruses from Diarrheic Beef and Dairy Calves using Monoclonal Antibodies in ELISA

1994 ◽  
Vol 6 (2) ◽  
pp. 175-181 ◽  
Author(s):  
A. Lucchelli ◽  
S. Y. Kang ◽  
M. K. Jayasekera ◽  
A. V. Parwani ◽  
D. H. Zeman ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
South Dakota ◽  
Dairy Herd ◽  
Dairy Calves ◽  
Enzyme Linked Immunosorbent Assay ◽  
Positive Stool ◽  
Field Samples ◽  
Group A Rotaviruses ◽  
Beef Herds ◽  
Group A

Group A bovine rotaviruses (BRV) have been identified worldwide as a major cause of diarrhea in the young of many species, including humans. Group A rotaviruses are classified into serotypes on the basis of the outer capsid proteins, VP7 (G types) and VP4 (P types). To date, there are 14 G types of group A rotaviruses, with G1, G6, G8, and G10 described for BRV isolates. In this study, G6- and G lo-specific monoclonal antibodies (MAbs) were used in an enzyme-linked immunosorbent assay (ELISA) for the G typing of BRV-positive stool samples from diarrheic beef and dairy calves from South Dakota, Ohio, Michigan, Nebraska, and Washington, USA, and Ontario, Canada. ELISA plates were coated using a broadly reactive VP7 MAb (Common 60) or with G6- or G10-specific MAbs. BRV-positive fecal samples were diluted and added to duplicate wells, followed by the addition of polyclonal guinea pig anti-group A rotavirus serum as the secondary antibody. Several reference G6 and G10 BRV strains as well as other G types previously reported in cattle (G1, G2, G3, G8) and BRV-negative samples were included as G type specificity and negative controls. From a total of 308 field samples analyzed, 79% (244/308) tested positive by the broadly reactive VP7 MAb; of these, 54% (131/244) were G6 positive, 14% (35/244) were G10 positive, 4% (9/244) were both G6 and G10 positive, and 28% (69/244) were G6 and G10 negative. The negative samples may represent additional or undefined serotypes. The 89 samples from South Dakota were further subdivided into samples from beef ( n = 43) or dairy ( n = 46) herds. G6 was more prevalent in beef herd samples (67%) than in dairy herd samples (47.5%). In addition, dairy herds had higher percentages of G10-positive samples (17.5%) G6-G10 double positives (10%), and untypable samples (25%) than did beef herds, in which the prevalence of G10 positive samples was 5.5%, G6-G10 double positives was 5.5%, and untypable samples was 22%. Application of the serotype ELISA for the analysis of additional BRV samples will provide further epidemiologic data on the distribution of BRV serotypes in beef or dairy cattle, an important consideration for the development of improved BRV vaccines.

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