Validation of the BioPRYN enzyme-linked immunosorbent assay for detection of pregnancy-specific protein-B (PSPB) and diagnosis of pregnancy in American bison (Bison bison )

2017 ◽  
Vol 52 (5) ◽  
pp. 791-797 ◽  
Author(s):  
DM Love ◽  
MT Mefford ◽  
JC Ramer
Keyword(s):  
Immunosorbent Assay ◽  
Specific Protein ◽  
Bison Bison ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein B

An Enzyme-Linked Immunosorbent Assay for the Rapid Detection of Staphylococcus aureus in Processed Foods

1994 ◽  
Vol 57 (3) ◽  
pp. 184-189 ◽  
Author(s):  
TSUNG C. CHANG ◽  
SU H. HUANG
Keyword(s):  
Staphylococcus Aureus ◽  
Immunosorbent Assay ◽  
Rapid Detection ◽  
Protein A ◽  
Large Degree ◽  
Microtiter Plate ◽  
Specific Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Processed Foods ◽  
Culture Methods

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the rapid detection of Staphylococcus aureus in foods. The assay was based on the detection of protein A which is a specific protein secreted by S. aureus. Following a 24-h incubation in a staphylococcal selective broth containing mannitol as the carbon source, the culture supematant was added to the microtiter plate coated with anti-protein A immunoglobulin G (IgG). After incubation, peroxidase-labeled anti-protein A IgG was used to produce the signal of antigen-antibody reaction. The sensitivity of the assay for protein A was 0.1 ng/ml. For 37 strains of S. aureus studied, all produced protein A, and the amount (13-1,100 ng/ml) of protein A secreted by different strains varied to a large degree. For another 57 strains (including 19 Staphylococcus spp.) of bacteria tested, two strains (5. capilis subsp. capitis CCRC 12161 and S. lentus CCRC 12926) produced very low amounts of protein A (0.6-1 ng/ml) after 24-h incubation. Staphylococcus aureus was detected by the ELISA in all of six samples of precooked foods naturally contaminated with the bacterium. Twenty-two processed foods artificially inoculated with S. aureus at levels of < 2 CFU/g and 10 to 20 CFU/g, respectively, were all positive by the ELISA. As compared to the conventional culture methods which take 5 to 6 days to complete, the ELISA can detect low numbers of S. aureus in processed foods with a total analytical time of only 28 h.

The Determination of IgG Subclass Concentrations in Serum by Enzyme-Linked Immunosorbent Assay: Establishment of Age-Related Reference Ranges for Cord Blood Samples, Children Aged 5–13 Years and Adults

1994 ◽  
Vol 31 (3) ◽  
pp. 245-248 ◽  
Author(s):  
Judith Miles ◽  
Pamela Riches
Keyword(s):  
Immunosorbent Assay ◽  
Specific Protein ◽  
Igg Subclasses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reference Ranges ◽  
Age Related ◽  
Normal Individuals ◽  
Coefficients Of Variation ◽  
Colour Development ◽  
Phosphate Buffered Saline

We describe a simple enzyme-linked immunosorbent assay for the measurement of immunoglobulin G (IgG) subclasses in serum. The microtitre plate is coated directly with diluted samples, standards or controls, blocked with phosphate-buffered saline containing 1% Tween to prevent non-specific protein binding to the plate, and each subclass assigned with specific mouse monoclonal antibody. The specific antibody is detected with alkaline-phosphatase linked anti-globulin and the colour development of the paranitrophenyl phosphate indicator reagent is measured at 405 nm until the highest concentration of standard gives an absorbance in excess of 1·4. Coefficients of variation measured at three concentration levels were less than 7% for within-assay variation and less than 10% for between-assay variation. Regression analysis of total IgG with the sum of the measured IgG subclasses gave a correlation coefficient of 0·932 ( P < 0·001). The assay has been used to establish age-related reference ranges for serum IgG subclasses; these are found to be slightly different from those established for radioimmunodiffusion methods. Our study does not confirm the previously reported absence of IgG4 in normal individuals.

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