Development of a double monoclonal antibody sandwich ELISA test forVerticillium dahliaeKleb.

Canine atopic dermatitis (CAD) is a pruritic allergic skin disease associated with IgE-mediated hypersensitivity. IgE is detected using Serum Allergen-Specific IgE test (SAT) in order to identify allergens. The present study aims to identify the environmental allergens in atopic dogs living in Northern Italy using SAT. The screening SAT (sSAT), using a monoclonal antibody cocktail-based ELISA to identify indoor and outdoor allergens, was performed. In all positive samples, an anti-IgE monoclonal antibody ELISA test was performed to extend panel of allergens. Out of 117 selected dogs, 69 were included in the study; 71% were positive and 29% were negative to sSAT. Among the 49 positive sSAT, 53% were positive for both indoor and outdoor, 38.8% only for indoor, and 8.2% only for outdoor allergens. This is the first study on the frequency of allergens involved in CAD in Italy using SAT. IgE hypersensitivity in atopic dogs of Northern Italy is usually associated with indoor allergens, primarily house dust mites. Among the outdoor allergens, an important role was played by Rumex acetosa. Polysensitization also commonly occurs. Therefore, since the numerous factors affect the IgE positivity in CAD, specific panels for geographical areas should be considered and re-evaluated at time intervals.

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Lactate Dehydrogenase Monoclonal Antibody Sandwich ELISA To Determine Cooking Temperature of Ground Beef

Journal of Agricultural and Food Chemistry ◽

10.1021/jf960327c ◽

1996 ◽

Vol 44 (12) ◽

pp. 4048-4051 ◽

Cited By ~ 7

Author(s):

A. Orta-Ramirez ◽

C. H. Wang ◽

M. M. Abouzied ◽

G. J. Veeramuthu ◽

J. F. Price ◽

...

Keyword(s):

Monoclonal Antibody ◽

Lactate Dehydrogenase ◽

Sandwich Elisa ◽

Ground Beef ◽

Cooking Temperature

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Detection of Mycoplasma agalactiae Antigen in Sheep and Goats by Monoclonal Antibody-Based Sandwich ELISA

Acta Veterinaria Brno ◽

10.2754/avb200473040461 ◽

2004 ◽

Vol 73 (4) ◽

pp. 461-464 ◽

Cited By ~ 1

Author(s):

D. Zendulková ◽

H. J. Ball ◽

A. Madanat ◽

P. Lány ◽

Z. Pospíšil

Keyword(s):

Monoclonal Antibody ◽

Sandwich Elisa ◽

Mycoplasma Agalactiae ◽

Sheep And Goats

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Development of a Sandwich Enzyme-Linked Immunosorbent Assay for Detection and Quantification of Clam Residues in Food Products

BioMed Research International ◽

10.1155/2021/6685575 ◽

2021 ◽

Vol 2021 ◽

pp. 1-9

Author(s):

Stef J. Koppelman ◽

Ashley L. Lardizabal ◽

Lynn Niemann ◽

Joe L. Baumert ◽

Steve L. Taylor

Keyword(s):

Sandwich Elisa ◽

Food Product ◽

Food Products ◽

Elisa Test ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Allergic Reactions ◽

Arctica Islandica ◽

Limit Of Quantification ◽

Detection And Quantification

Seafood is a frequent cause of allergic reactions to food globally. The presence of undeclared trace amounts of clam can cause allergic reactions in sensitive individuals. Limited tools are available to test food products for the presence of traces of clam. We report on the development of a sandwich ELISA that can detect and quantify clam protein in food. Antisera against a mix of two commercially important clam species, Atlantic Surf (Spisula solidissima) and ocean quahog (Arctica islandica), were raised in rabbit and sheep. A sandwich ELISA was constructed with this antisera, and sensitivity and specificity were evaluated. Also, model food products spiked with clam protein were analyzed to assess the performance of the ELISA. Comparison was made with a commercially available ELISA for crustacea. The lower limit of quantification of the sandwich ELISA is 2.5 ppm clam protein in food samples, allowing the detection of low amounts of clam that may trigger a reaction in clam allergic patients. The sandwich ELISA was highly specific with cross-reactivity only noted for other molluscan shellfish (mussel and scallop). Clam protein in tomato juice and potato cream soup was detected well with recoveries ranging from 65 to 74% and from 74 to 113%, respectively. However when potato cream soup was retorted, the recover fell to 20%, imposing the risk of underestimating the clam content of a food product. A commercially available crustacean ELISA test was not suitable to detect clam protein. The sandwich ELISA described here is suitable for detection and quantification of clam protein in food products. Care should be taken with food products that have been retorted as the results may be underestimated.

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Development of a monoclonal antibody against the left wing of ciguatoxin CTX1B: Thiol strategy and detection using a sandwich ELISA

Toxicon ◽

10.1016/j.toxicon.2012.04.347 ◽

2012 ◽

Vol 60 (3) ◽

pp. 348-357 ◽

Cited By ~ 21

Author(s):

Takeshi Tsumuraya ◽

Katsutoshi Takeuchi ◽

Shuji Yamashita ◽

Ikuo Fujii ◽

Masahiro Hirama

Keyword(s):

Monoclonal Antibody ◽

Sandwich Elisa ◽

Left Wing

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Comparative Diagnosis of Serum IgG1 and Coproantigen ELISA for Fasciolosis Detection of Goats in Mexico

BioMed Research International ◽

10.1155/2016/3860928 ◽

2016 ◽

Vol 2016 ◽

pp. 1-7 ◽

Cited By ~ 1

Author(s):

Abel Villa-Mancera ◽

Pedro Molina-Mendoza ◽

Karina Hernández-Guzmán ◽

Jaime Olivares-Pérez ◽

Jorge Sarracent-Pérez ◽

...

Keyword(s):

Serological Test ◽

Sandwich Elisa ◽

Correlation Coefficients ◽

Economic Loss ◽

Elisa Test ◽

Current Status ◽

Antigen Elisa ◽

Faecal Samples ◽

Diagnostic Sensitivity And Specificity ◽

Serum Elisa

The objective of present study was to determine the prevalence of natural caprine fasciolosis in the Mixteca region of Mexico using coproantigen and serum IgG1 ELISA tests for comparative purposes. A total of 1070 serum and faecal samples were analyzed for IgG1 antibodies and coproantigens, using ELISA with E/S products as antigen and a monoclonal antibody-based sandwich ELISA. Prevalence of 73.46% was found using the serological ELISA and a percentage of 77.20 was found for coproantigen ELISA. The diagnostic sensitivity and specificity for serum ELISA were 86.7% and 96.4%, and for the coproantigen ELISA they were 93.1% and 97.8%, respectively. The seropositive samples were further categorized as low, medium, or high positivity. Results show a great proportion of low and medium positive goats when the serum ELISA test was used. Correlation coefficients between coproantigens and seropositivity were statistically significant (P<0.01) for low seropositivity (r=0.93) and medium seropositivity (r=0.84). The accuracy of faecal antigen ELISA was higher compared to indirect ELISA serological test. Two ELISAs were shown to be useful for demonstrating the current status ofF. hepaticainfection in the endemic areas and can be employed in studies on epidemiology as well as anthelmintics treatment for preventing economic loss and the risk of transmission to humans.

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PRRSV Vaccine Strain-Induced Secretion of Extracellular ISG15 Stimulates Porcine Alveolar Macrophage Antiviral Response against PRRSV

Viruses ◽

10.3390/v12091009 ◽

2020 ◽

Vol 12 (9) ◽

pp. 1009

Author(s):

Hongbin Liu ◽

Bingjun Shi ◽

Zhigang Zhang ◽

Bao Zhao ◽

Guangming Zhao ◽

...

Keyword(s):

Monoclonal Antibody ◽

Vaccine Strain ◽

Sandwich Elisa ◽

Respiratory Syndrome Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Porcine Alveolar Macrophage ◽

Type I ◽

Swine Industry ◽

Host Interactions ◽

Isg15 Expression

Porcine reproductive and respiratory syndrome virus (PRRSV) has disrupted the global swine industry since the 1980s. PRRSV-host interactions are largely still unknown but may involve host ISG15 protein. In this study, we developed a monoclonal antibody (Mab-3D5E6) specific for swine ISG15 (sISG15) by immunizing mice with recombinant sISG15. A sandwich enzyme-linked immunosorbent assay (ELISA) incorporating this sISG15-specific Mab was developed to detect sISG15 and provided a lower limit of sISG15 detection of 200 pg/mL. ELISA results demonstrated that infection of porcine alveolar macrophages (PAMs) with low-virulence or attenuated PRRSV vaccine strains induced intracellular ISG15 expression that was independent of type I IFN production, while PAMs infection with a PRRSV vaccine strain promoted extracellular ISG15 secretion from infected PAMs. Conversely, the addition of recombinant sISG15 to PAMs mimicked natural extracellular ISG15 effects whereby sISG15 functioned as a cytokine by activating PAMs. Once activated, PAMs could inhibit PRRSV replication and resist infection with PRRSV vaccine strain TJM. In summary, a sandwich ELISA incorporating homemade anti-ISG15 Mab detected ISG15 secretion induced by PAMs infection with a PRRSV vaccine strain. Recombinant ISG15 added to cells exhibited cytokine-like activity that stimulated PAMs to assume an anti-viral state that enabled them to inhibit PRRSV replication and resist viral infection.

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Monoclonal antibodies specific for prothrombin fragment 1.2 and their use in a quantitative enzyme-linked immunosorbent assay

Clinical Chemistry ◽

10.1093/clinchem/39.4.583 ◽

1993 ◽

Vol 39 (4) ◽

pp. 583-591 ◽

Cited By ~ 13

Author(s):

M J Hursting ◽

B T Butman ◽

J P Steiner ◽

B M Moore ◽

M C Plank ◽

...

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Human Plasma ◽

Sandwich Elisa ◽

Peptide Sequence ◽

Carboxyl Terminus ◽

Enzyme Linked Immunosorbent Assay ◽

Thrombotic Risk ◽

Prothrombin Fragment ◽

Amino Terminal

Abstract Prothrombin fragment 1.2 (F1.2) is an activation peptide generated during a critical event of blood coagulation, the conversion of prothrombin to thrombin. As a marker of thrombin generation, F1.2 has clinical potential in assessing thrombotic risk and monitoring anticoagulant therapy. In developing a highly specific, monoclonal antibody-based immunoassay of human plasma F1.2, we generated six murine anti-F1.2 monoclonal antibodies, using as immunogen a synthetic peptide (sequence: CGSD-RAIEGR) similar to the unique carboxyl terminus of F1.2. Each antibody bound F1.2 but not prothrombin. Epitope mapping studies with one antibody (5-3B) showed that optimum binding required six to eight amino acids plus a terminal arginine to emulate the F1.2 carboxyl terminus. A quantitative sandwich ELISA for human plasma F1.2 was configured with monoclonal antibody 5-3B as the capture antibody and peroxidase-labeled polyclonal antibodies to the F1.2 amino-terminal region as detector antibodies. Calibrators were prepared by adding purified F1.2, 0-10 nmol/L, to F1.2-depleted plasma. Assay characteristics included the following: mean (+/- SD) analytical recovery of 98% +/- 13%; no interference from lipemia, hemolysis, icterus, or thrombolytic agents; 0.08 nmol/L sensitivity; and mean intra- and interassay imprecision (three lots) < 12% at both low and high concentrations of F1.2.

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