Enzyme-linked immunosorbent assay for 17 alpha-hydroxyprogesterone in dried blood spotted on filter paper.

1987 ◽  
Vol 33 (6) ◽  
pp. 761-764 ◽  
Author(s):  
M Maeda ◽  
H Arakawa ◽  
A Tsuji ◽  
Y Yamagami ◽  
A Isozaki ◽  
...  
Keyword(s):  
Detection Limit ◽  
Immunosorbent Assay ◽  
Filter Paper ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Adrenal Hyperplasia ◽  
Bovine Serum Albumin Conjugate ◽  
Enzyme Conjugate ◽  
Sensitivity Specificity ◽  
Microwell Plate

Abstract In this rapid, cost-effective, enzyme-linked immunosorbent assay for 17 alpha-hydroxyprogesterone (17-OHP) eluted from dried blood spotted on filter paper, second antibody is coated onto the microwell plate and horseradish peroxidase (EC 1.11.1.7) is the label enzyme. Antiserum to 17-OHP was prepared by using 4-(2-carboxymethylthio)-17-OHP-bovine serum albumin conjugate as immunogen. Enzyme conjugate was prepared from 4-(2-carboxymethylthio)-17-OHP and peroxidase. The blood spots are assayed in the microwells without extraction or centrifugation steps. The detection limit of the assay is 1 microgram/L, equivalent to 3.5 pg (10.6 fmol) per disc. Intra- and interassay CVs at two steroid concentrations (7.38 and 22.79 micrograms/L) ranged from 3.74 to 11.90% (n = 5), and 9.49 and 9.83% (n = 5), respectively. Results correlated well (r = 0.91) with those of a fluorescence enzyme immunoassay. The sensitivity, specificity, and precision of this method make it potentially useful in the mass screening of neonates for congenital adrenal hyperplasia.

scholarly journals Easy Enzyme-Linked Immunosorbent Assay for Spectinomycin in Chicken Plasma

1996 ◽  
Vol 79 (2) ◽  
pp. 426-430 ◽  
Author(s):  
Touichi Tanaka ◽  
Hideharu Ikebuchi ◽  
Jun-Ichi Sawada ◽  
Mariko Okada ◽  
Yasumasa Kido
Keyword(s):  
Detection Limit ◽  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Immunosorbent Assay ◽  
Bovine Serum ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Carrier Protein ◽  
Bovine Serum Albumin Conjugate

Abstract An easy, sensitive, competitive indirect enzyme- linked immunosorbent assay (CI-ELISA) for specti nomycin in chicken plasma was developed. Preparation of a spectinomycin-bovine serum albumin conjugate in which the hapten is linked to the carrier protein through the C-4 position is described. Antibodies raised against antigens in rabbits had excellent specificity for spectinomycin, exhibiting a cross-reactivity of 44.0% with dihydrospectinomy-cin and 13.8% with tetrahydrospectinomycin. No cross-reactivity was observed with other antibiotics. The detection limit of the CI-ELISA was 2 ng/mL (equivalent into 40 ng/mL undiluted chicken plasma) spectinomycin. Known amounts (0.1-100 μg/mL) of spectinomycin were added to chicken plasma and then analyzed. Average recoveries were 97-110%. This procedure may be used without prior extraction of samples.

Enzyme-Linked Immunosorbent Assay for Streptomycin in Animal Feeds

2013 ◽  
Vol 651 ◽  
pp. 280-283
Author(s):  
Hui Ying Zhang ◽  
Jun Ping Wang
Keyword(s):  
Detection Limit ◽  
Bovine Serum Albumin ◽  
Immunosorbent Assay ◽  
Polyclonal Antibody ◽  
Bovine Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Animal Feeds ◽  
Inhibition Concentration ◽  
Coefficients Of Variation ◽  
Bovine Serum Albumin Conjugate

Polyclonal antibody against streptomycin was prepared by using a streptomycin–bovine serum albumin conjugate for the immunization of rabbits. Using this antibody, we developed quantitative assays for streptomycin by means of an indirect competitive enzyme-linked immunosorbent assay (icELISA). Fifty percent inhibition concentration (IC50) for the antibody was 3.6 ng/ml. The detection limit was 0.4 ng/ml. The average of recoveries for all samples was 86.14% and the coefficients of variation of intra- and inter-assays were below 18%. The detection limit using the kit was 15 ng/ml in animal feeds.

An enzyme-linked immunosorbent assay for the detection of Rathayibacter toxicus, the bacterium involved in annual ryegrass toxicity, in hay

2006 ◽  
Vol 57 (7) ◽  
pp. 731 ◽  
Author(s):  
A. M. Masters ◽  
A. R. Gregory ◽  
R. J. Evans ◽  
J. E. Speijers ◽  
S. S. Sutherland
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Specific Antigen ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Annual Ryegrass ◽  
Capture Assay ◽  
Large Numbers ◽  
Soluble Polysaccharide ◽  
Annual Ryegrass Toxicity

An enzyme-linked immunosorbent assay (ELISA) for Rathayibacter toxicus is described. The development of a monoclonal antibody for a specific antigen from R. toxicus and a polyclonal antibody raised against the same R. toxicus preparation enabled a capture assay format. The assay is specific for a soluble polysaccharide produced by the bacterium and was found to be sensitive enough to detect antigen equivalent to less than one gall per kilogram of hay. The applicability of the assay to samples of pasture or hay is demonstrated. Cost-effective testing of large numbers of samples for the presence of R. toxicus is possible with the ELISA. This will assist stockowners, hay producers, and hay exporters in the management of the risk of annual ryegrass toxicity.

Aqueous two-phase system antibody confinement enables cost-effective analysis of protein analytes by sandwich enzyme-linked immunosorbent assay with minimal optical crosstalk

The Analyst ◽  
2020 ◽  
Vol 145 (16) ◽  
pp. 5458-5465 ◽  
Author(s):  
Maia Kvas ◽  
Alyne G. Teixeira ◽  
Beatrice Chiang ◽  
John P. Frampton
Keyword(s):  
Immunosorbent Assay ◽  
Low Cost ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Phase System ◽  
Two Phase ◽  
Aqueous Two Phase System ◽  
Optical Crosstalk ◽  
Effective Analysis ◽  
Protein Analytes

An aqueous two-phase system was used to reduce reagent volumes and optical crosstalk for a low-cost single sandwich enzyme-linked immunoassay.

scholarly journals Neonatal screening for congenital adrenal hyperplasia

2004 ◽  
pp. U71-U75 ◽  
Author(s):  
HJ van der Kamp ◽  
JM Wit
Keyword(s):  
Birth Weight ◽  
Congenital Adrenal Hyperplasia ◽  
Gestational Age ◽  
Filter Paper ◽  
Early Recognition ◽  
Enzyme Linked Immunosorbent Assay ◽  
Adrenal Hyperplasia ◽  
Salt Wasting ◽  
Specificity And Sensitivity ◽  
The Usa

Congenital adrenal hyperplasia (CAH) is well suited for newborn screening, as it is a common and potentially fatal disease which can be easily diagnosed by a simple hormonal measurement in blood. Moreover, early recognition and treatment can prevent severe salt wasting, dehydration and death and shorten the time of male sex assignment in virilised females.In screening programmes, 17alpha-hydroxyprogesterone (17OHP) is measured in filter paper blood spots obtained by a heel puncture preferably between 2 and 4 days after birth. Three assay techniques are utilised for initial screening: radio-immunoassay (USA), enzyme-linked immunosorbent assay (Japan) and time-resolved fluoro-immunoassay (Europe). Preterm newborns have higher 17OHP concentrations in serum than babies born at term. Therefore, cut-off levels are based on gestational age (in Japan and Europe) or on birth weight (in the USA). There is a considerable variation in cut-off levels from one programme to another. This is most likely due to the different antibodies and reagents used, varying thickness and density of filter paper used for sample collection and, most significantly, the characteristics of the reference population (in terms of birth weight and gestational age).More than 30 million newborns have been screened. The prevalence of CAH in the USA and Europe is approximately 1:15 000-16 000, and slightly lower in Japan (1:19 000). In general, severe salt wasting can be prevented, but there is a remarkable variation in the number of false-positives and false-negatives among the various programmes. Ongoing refinement of cut-off levels is needed to improve specificity and sensitivity.

scholarly journals Comparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay (ELISA)

2017 ◽  
Vol 8 ◽  
pp. 244-253 ◽  
Author(s):  
Paula Ciaurriz ◽  
Fátima Fernández ◽  
Edurne Tellechea ◽  
Jose F Moran ◽  
Aaron C Asensio
Keyword(s):  
Gold Nanoparticles ◽  
Detection Limit ◽  
Immunosorbent Assay ◽  
Wheat Gluten ◽  
Diagnostic Technique ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rabbit Igg ◽  
Enzyme Molecules ◽  
Main Components ◽  
Limit Sensitivity

The enzyme-linked immunosorbent assay (ELISA) technique is based on the specific recognition ability of the molecular structure of an antigen (epitope) by an antibody and is likely the most important diagnostic technique used today in bioscience. With this methodology, it is possible to diagnose illness, allergies, alimentary fraud, and even to detect small molecules such as toxins, pesticides, heavy metals, etc. For this reason, any procedures that improve the detection limit, sensitivity or reduce the analysis time could have an important impact in several fields. In this respect, many methods have been developed for improving the technique, ranging from fluorescence substrates to methods for increasing the number of enzyme molecules involved in the detection such as the biotin–streptavidin method. In this context, nanotechnology has offered a significant number of proposed solutions, mainly based on the functionalization of nanoparticles from gold to carbon which could be used as antibody carriers as well as reporter enzymes like peroxidase. However, few works have focused on the study of best practices for nanoparticle functionalization for ELISA enhancement. In this work, we use 20 nm gold nanoparticles (AuNPs) as a vehicle for secondary antibodies and peroxidase (HRP). The design of experiments technique (DOE) and four different methods for biomolecule loading were compared using a rabbit IgG/goat anti-rabbit IgG ELISA model (adsorption, directional, covalent and a combination thereof). As a result, AuNP probes prepared by direct adsorption were the most effective method. AuNPs probes were then used to detect gliadin, one of the main components of wheat gluten, the protein composite that causes celiac disease. With this optimized approach, our data showed a sensitivity increase of at least five times and a lower detection limit with respect to a standard ELISA of at least three times. Additionally, the assay time was remarkably decreased.

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