Journal of immunological methods.
Publisher:
North-Holand,. Amsterdam : Elsevier
Frequency: Twenty four no. a year
Country: Netherlands
Language: English
Author(s):
Association of Medical Laboratory Immunologists.
Start Year:1971 -
ISSN:
0022-1759 (Print)
1872-7905 (Electronic)
0022-1759 (Linking)
1872-7905 (Electronic)
0022-1759 (Linking)
Impact Factor
2.2
2022
| NLM ID: | 1305440 |
| (DNLM): | J23720000(s) |
| (OCoLC): | 01783876 |
| Coden: | JIMMBG |
| LCCN: | 72617482 |
| Classification: | W1 JO676K |
Novel bridge multi-species ELISA for detection of SARS-CoV-2 antibodies. Considering the course of the current SARS-CoV-2 pandemic, it is important to have serological tests for monitoring humoral immune response against SARS-CoV-2 infection and vaccination. Herein we describe a novel bridge enzyme-linked immunosorbent assay (b-ELISA) for SARS-CoV-2 antibodies detection in human and other species, employing recombinant Spike protein as a unique antigen, which is produced at high scale in insect larvae. Eighty two human control sera/plasmas and 169 COVID-19 patients' sera/plasmas, confirmed by rRT-PCR, were analyzed by the b-ELISA assay. In addition, a total of 27 a...
Evaluation of refractometry methods for estimating passive immunity status in neonatal foals. The objective of this cross-sectional study was to determine the accuracy of the digital Brix and serum total protein (TP) refractometers for estimating different passive immunity status in neonatal foals. In total, 18- to 40-h old purebred Arabian foals (n = 185) were used. Serum TP concentrations, total solid percentages and IgG concentrations were measured with a digital serum TP refractometer, digital Brix refractometer and the gold standard radial immunodiffusion (RID) assay, respectively. Correlation coefficients were calculated between the refractometer and RID assay results. A receiv...
Development and evaluation of recombinant antigen and monoclonal antibody based competition ELISA for the sero- surveillance of surra in animals. Trypanosoma evansi, a haemoflagellated protozoan parasite, is responsible for chronic as well as the acute debilitating disease called surra in a wide range of herbivores and carnivores including domestic and wild animals. Since the parasite is having wide host range, there is a need for diagnostic test which can detect the T. evansi specific antibody in different species of animals for generating sero-surveillance data. In the present study we developed and evaluated competitive enzyme immunoassay using monoclonal antibodies (MAbs) raised against recombinant variable surface glycoprotein (rVS...
Optimizing selection of large animals for antibody production by screening immune response to standard vaccines. Antibodies made in large animals are integral to many biomedical research endeavors. Domesticated herd animals like goats, sheep, donkeys, horses and camelids all offer distinct advantages in antibody production. However, their cost of use is often prohibitive, especially where poor antigen response is commonplace; choosing a non-responsive animal can set a research program back or even prevent experiments from moving forward entirely. Over the course of production of antibodies from llamas, we found that some animals consistently produced a higher humoral antibody response than others, even t...
Validation of quantitative polymerase chain reaction assays for measuring cytokine expression in equine macrophages. The study of the equine immune system and inflammatory responses, by measuring cytokine expression, can provide important insight into disease pathogenesis in the horse. A set of quantitative real-time polymerase chain reaction (QPCR) assays for the equine cytokines IL-1alpha, IL-1beta, IL-6, IL-8 and TNF-alpha were validated using QPCR primers and probes which were generated for the equine IL-1alpha, IL-1beta, IL-6, IL-8, TNF-alpha and 18S genes. Amplification efficiency, intra-assay and inter-assay variation were determined using 10-fold dilutions of plasmid for each gene. Under these condit...
Area under the curve calculations as a tool to compare the efficacy of equine influenza vaccines–a retrospective analysis of three independent field trials. Using the area under the curve (AUC) concept as is commonly used in pharmaceutical bioequivalence studies, the bioequivalence of three equine influenza vaccines was demonstrated. A retrospective analysis was performed using this technique on data generated in three trials in which each of the three vaccines had been used. In total, data from 63 pony and horse foals were used. The AUC of the single radial hemolysis (SRH) titres against Influenza A/equi-1/Prague/56 (Pr/56), A/equi-2/Newmarket-1/93, and A/equi-2/Suffolk/89 (Suf/89) were calculated for each horse. It was concluded that calculation...
Quantitative measurement of equine cytokine mRNA expression by polymerase chain reaction using target-specific standard curves. Quantification of cytokine mRNA using reverse transcription coupled with the polymerase chain reaction (RT-PCR) has become a corner stone of the study of cytokine regulation. Quantitative competitive RT-PCR (QCRT-PCR) is commonly accepted as a reliable method for quantifying differences in mRNA levels but is both labor- and reagent-intensive. A noncompetitive polymerase chain reaction method that utilizes cytokine-specific, plasmid-derived, standard curves was developed for the quantification of equine cytokine mRNA. The assay can be performed on minute samples of cellular material, utilizes s...
General method for the detection and in vitro expansion of equine cytolytic T lymphocytes. Equine immunological research is hindered by the lack of a simple yet reliable general protocol by which to assay CTL activity specific for viral or parasitic antigens. We present here the first comprehensive analysis of the parameters necessary to reliably culture equine T cells and to analyze the antigen specific cytolytic activity of T lymphocytes utilizing the equine infectious anemia virus (EIAV) infection of outbred ponies as a source for in vivo primed T lymphocytes. Effective long-term in vitro culture of equine T cells was determined to require minimally 200 U/ml of recombinant human ...
Development of an ELISA using a universal method of enzyme-labelling drug-specific antibodies. Part I: Detection of dexamethasone in equine urine. The development, validation, and application of an ELISA for dexamethasone in equine urine is described. The drug-protein conjugate was immobilised in microtitre plate wells and antiserum raised against the same drug-protein conjugate was allowed to compete with sample or standard drug and the immobilised drug-protein conjugate. The proportion of antiserum binding to the immobilised drug-protein conjugate was detected using a biotinylated protein G/extravidin-alkaline phosphatase complex in situ and measurement of the substrate product. The method was used to detect the presence of drug-derive...
A versatile synthetic peptide-based ELISA for identifying antibody epitopes. A simple, versatile and very inexpensive procedure for cross-linking synthetic peptides to the polystyrene surfaces of micro-well assay plates for use in ELISA was developed. The method is based on the use of poly-L-lysine (PLL) as the anchor protein for synthetic peptides which were then easily and covalently linked to the PLL using glutaraldehyde. The synthetic peptides used for the study were based on the amino acid sequence of the equine infectious anemia virus (EIAV) envelope sequence and evaluated as antigens in an ELISA designed to detect antibodies in serum of EIAV-infected horses and ...
Biotin-labeled antigen sandwich enzyme-linked immunosorbent assay (BLA-S-ELISA) for the detection of Japanese encephalitis antibody in human and a variety of animal sera. A biotin-labeled antigen (BLA) was adapted to a sandwich enzyme-linked immunosorbent assay (S-ELISA) for detection of Japanese encephalitis (JE) antibody in a variety of animal sera. JE antigen was fixed on the wells of a microplate and became bound to the specific antibody which could react with a peroxidase-labeled avidin conjugate and azino-di-(3-ethylbenzthiazolin sulfonic acid) (ABTS) as a substrate. The BLA-S-ELISA could simultaneously detect JE antibody in all hemagglutination inhibition (HI) positive sera from man, swine, monkey, horse, cattle, rabbit, rat, mouse and pigeon by using th...
Simultaneous preparation of mononuclear and polymorphonuclear leucocytes from horse blood on Ficoll-Hypaque medium. Results presented show that highly purified populations of mononuclear (MN) and polymorphonuclear (PMN) leucocytes can be obtained from horse blood by a procedure similar to that previously described for the separation of these leucocytes from human blood. This involved centrifugation of horse blood on a Ficoll-Hypaque medium with a density of 1.095 g/ml. The procedure required approximately 1 h for completion and resulted in the simultaneous preparation of MN (greater than 98% purity) and PMN (greater than 96% purity) leucocytes. Cell viability exceeded 95% and cells retained immunological fu...
An enzyme immunoassay (EIA) for progesterone in horse plasma. A simple enzyme immunoassay (EIA) for the measurement of progesterone is described. Antibody against 11-OH-hemisuccinate-BSA is bound to polystyrene tubes. 11-OH-hemisuccinyl-beta-D-galactosidase is used as enzyme-coupled antigen and methylumbelliferyl-beta-D-galactoside as substrate. Concentrations down to 0.156 ng/ml plasm or amounts of 93 pg/tube are detectable. Probit analysis gave a linear relationship between log concentration and percentage of binding. A comparison of EIA and radioimmunoassay gave a correlation coefficient of 0.81. The assay is sufficiently sensitive to estimate progest...