PREGNANCY DIAGNOSIS IN OKAPI (OKAPIA JOHNSTONI) USING BIOPRYN ENZYME-LINKED IMMUNOSORBENT ASSAY FOR DETECTION OF PREGNANCY-SPECIFIC PROTEIN-B (PSPB)

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Matthew E. Kinney ◽  
Josh R. Branen
Keyword(s):  
Immunosorbent Assay ◽  
Specific Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Pregnancy Diagnosis ◽  
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.

75 COMPARISON OF THE DG29TM ENZYME-LINKED IMMUNOSORBENT ASSAY KIT COMPARED WITH TRANSRECTAL ULTRASONOGRAPHY FOR EARLY PREGNANCY DIAGNOSIS FOLLOWING TRANSFER OF JAPANESE BLACK CATTLE EMBRYOS

2012 ◽  
Vol 24 (1) ◽  
pp. 150 ◽  
Author(s):  
Y. Nakamura ◽  
M. Urakawa ◽  
A. Ideta ◽  
A. Shirasawa ◽  
Y. Oono ◽  
...  
Keyword(s):  
Negative Predictive Value ◽  
Clinical Application ◽  
Immunosorbent Assay ◽  
Early Pregnancy ◽  
Predictive Value ◽  
Transrectal Ultrasonography ◽  
Enzyme Linked Immunosorbent Assay ◽  
Japanese Black Cattle ◽  
Elisa Kit ◽  
Pregnancy Diagnosis

In commercial embryo transfer industry, accurate early detection of pregnant and nonpregnant cows plays a key role in achieving an optimum calving-to-conception interval. The DG29TM enzyme-linked immunosorbent assay (ELISA) kit (Conception, Animal Reproduction Technologies, Beaumont, Canada) measures the level of pregnancy-related glycoproteins in blood that are linked to pregnancy. Here, we compared the DG29 kit with transrectal ultrasonography (TU) to evaluate the possibility of the clinical application of the ELISA kit for early pregnancy diagnosis. Embryos recovered from superovulated Japanese Black cows were transferred to 110 recipient Holstein heifers on Day 6 to 8 of the oestrous cycle (oestrus = Day 0). Pregnancy was diagnosed between Day 29 and Day 40 by TU with a 5.0/7.5-MHz linear array transducer (Tringa, Pie Medical Equipment B.V., Maastricht, the Netherlands). Blood samples were collected from the tail vein or artery into vacuum serum tubes after TU and serum was separated and stored at –30°C until the ELISA was performed. The ELISA results were interpreted as positive (pregnant, >1000 pg mL–1) or negative (nonpregnant, <300 pg mL–1). Ninety-nine of the 110 heifers were judged as pregnant or nonpregnant by TU. Seventy-six of the 99 heifers were judged as pregnant, in which fetuses were visualised clearly in the uterine horn. The following measures, sensitivity, specificity, predictive value and accuracy of pregnancy outcomes based on the ELISA results, were assessed by comparing with those based on the definite TU results. The values for sensitivity and specificity were 100% (76/76) and 91.3% (21/23), the positive and negative predictive value were 97.4% (76/78) and 100% (21/21), respectively and accuracy was 98.0% (97/99). On the other hand, chorioallantoic fluids in 11 of the 110 heifers were detected by TU around Day 30 of gestation, but fetuses were not identified or were unclearly visualised, which suggests that the embryos died during the peri-implantation period after transfer. Ten of the 11 heifers were classified as pregnant by the ELISA, but only 3 heifers were identified as pregnant with reexamination by later TU, which indicates that the pregnancy-related glycoproteins residue from embryo mortality was detected by the DG29 kit. However, the negative predictive value of the DG29 kit was 100% in this study. In conclusion, except for early embryonic death, the DG29 kit was highly accurate and suitable for clinical application in early pregnancy determination following transfer of Japanese Black cattle embryos.

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