Monoclonal Antibodies and an Indirect ELISA for Detection of Psychrotrophic Bacteria in Refrigerated Milk

1997 ◽  
Vol 60 (1) ◽  
pp. 23-27 ◽  
Author(s):  
ROSALBA GUTIÉRREZ ◽  
ISABEL GONZÁLEZ ◽  
TERESA GARCÍA ◽  
ESTER CARRERA ◽  
BERNABÉ SANZ ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Horseradish Peroxidase ◽  
Indirect Elisa ◽  
Detection Threshold ◽  
Microtiter Plate ◽  
Live Cells ◽  
Elisa Assay ◽  
Psychrotrophic Bacteria ◽  
Milk Samples ◽  
Elisa Format

Monoclonal antibodies generated against live cells of Pseudomonas fluorescens have been used in an indirect ELISA format for the detection of Pseudomonas spp. and related psychrotrophic bacteria in refrigerated milk. The immunorecognition of monoclonal antibodies adsorbed to bacteria bound to the wells of a microtiter plate was performed with rabbit anti-mouse immunoglobulins conjugated to horseradish peroxidase. Subsequent enzymic conversion of the substrate resulted in distinct absorbance differences when assaying milk samples containing psychrotrophic bacteria in the range 105 to 109 CFU ml−1. The detection threshold for the ELISA assay developed in this work is 105 CFU ml−1.

Immunostick Colorimetric Assay for Detection of Pseudomonas spp. in Refrigerated Meat and Milk

1997 ◽  
Vol 60 (8) ◽  
pp. 908-911 ◽  
Author(s):  
ROSALBA GUTIÉRREZ ◽  
TERESA GARCÍA ◽  
ISABEL GONZÁLEZ ◽  
BERNABÉ SANZ ◽  
PABLO E. HERNÁNDEZ ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Pseudomonas Fluorescens ◽  
Colorimetric Assay ◽  
Detection Threshold ◽  
Live Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Pseudomonas Spp ◽  
Milk Samples ◽  
Elisa Format

Monoclonal antibodies generated against live cells of Pseudomonas fluorescens have been used in an immunostick enzyme-linked immunosorbent assay (ELISA) format for the detection of Pseudomonas spp. in refrigerated meat and milk. The detection threshold for the immunostick ELISA assay developed in this work is 104 CPU cm−2 for meat and 105 CPU ml−1 for milk samples.

Monoclonal Antibody Detection of Porcine Meat

1994 ◽  
Vol 57 (2) ◽  
pp. 146-149 ◽  
Author(s):  
P. MORALES ◽  
T. GARCÍA ◽  
I. GONZÁLEZ ◽  
R. MARTÍN ◽  
B. SANZ ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Bovine Serum Albumin ◽  
Horseradish Peroxidase ◽  
Indirect Elisa ◽  
Cross Reactivity ◽  
Antibody Detection ◽  
Muscle Proteins ◽  
Elisa Format ◽  
Soya Proteins

A stable hybridoma cell line (DD9) has been produced secreting a monoclonal antibody specific for porcine muscle proteins. The DD9 monoclonal antibody (mAb) failed to show a significant cross-reactivity when tested against beef, horse, chicken, and soya proteins, as well as bovine caseins, gelatin, and bovine serum albumin. The DD9 mAb was further used in an indirect ELISA format for detection of defined amounts of porcine meat (1–100%) in beef and chicken meat mixtures immobilized on 96-well plates. Immunorecognition of monoclonal antibodies adsorbed to porcine meat was made with rabbit anti-mouse immunoglobulins conjugated to the enzyme horseradish peroxidase.

scholarly journals Development and Application of a Novel Rapid and Throughput Method for Broad-Spectrum Anti-Foodborne Norovirus Antibody Testing

2021 ◽  
Vol 12 ◽  
Author(s):  
Yueting Zuo ◽  
Liang Xue ◽  
Junshan Gao ◽  
Yingyin Liao ◽  
Yueting Jiang ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Broad Spectrum ◽  
High Throughput Screening ◽  
Indirect Elisa ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Antibody Testing ◽  
Coefficients Of Variation ◽  
P Proteins

Foodbone norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Candidate vaccines are being developed, however, no licensed vaccines are currently available for managing NoV infections. Screening for stimulated antibodies with broad-spectrum binding activities can be performed for the development of NoV polyvalent vaccines. In this study, we aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA) for testing the broad spectrum of anti-NoV antibodies. Capsid P proteins from 28 representative NoV strains (GI.1–GI.9 and GII.1–GII.22 except GII.11, GII.18, and GII.19) were selected, prepared, and used as coating antigens on one microplate. Combined with incubation and the horseradish peroxidase chromogenic reaction, the entire process for testing the spectrum of unknown antibodies required 2 h for completion. The intra-assay and inter-assay coefficients of variation were less than 10%. The new method was successfully performed with monoclonal antibodies and polyclonal antibodies induced by multiple antigens. In conclusion, the indirect ELISA assay developed in this study had a good performance of reliability, convenience, and high-throughput screening for broad-spectrum antibodies.

Development and evaluation of an immunoenzymometric assay for the chicken progesterone receptor

1991 ◽  
Vol 129 (2) ◽  
pp. 189-196 ◽  
Author(s):  
M. K. Bläuer ◽  
P. J. Tuohimaa ◽  
P. J. Vilja
Keyword(s):  
Monoclonal Antibodies ◽  
Small Intestine ◽  
Horseradish Peroxidase ◽  
Progesterone Receptor ◽  
Ligand Binding ◽  
Binding Assay ◽  
Ligand Binding Assay ◽  
Coefficients Of Variation ◽  
First Time ◽  
Determination Range

ABSTRACT A specific and sensitive immunoenzymometric assay (IEMA) was developed for measuring the quantity of chicken progesterone receptor (PR) in tissue cytosol. The assay uses two monoclonal antibodies to the PR. One is used to capture the PR. The second (labelled with biotin) reacts first with the captured receptor and subsequently with avidin-labelled horseradish peroxidase to provide an enzymatic end-point. The method has a determination range from 0·3 to 60 pmol/l. Intra- and interassay coefficients of variation were 3·7% and 9·0% respectively. The assay can be performed with equal results as a rapid (3 h) or an overnight procedure. The IEMA is convenient, especially for signal measurement and the calculation of results. No ultracentrifugation of samples is needed, since the IEMA can be performed on low-speed cytosol samples. Assay results correlated well (r = 0·927) with those obtained by the conventional ligand-binding assay used in our laboratory. Similar results were obtained with the IEMA and the ligand-binding assay after exposure of cytosol samples to increased temperatures: at 20 °C the PR remained stable for the 4-h period examined, whereas at 37 °C almost complete degradation of the PR was observed in 30 min. Being more than 100 times as sensitive as the ligand-binding assay, the IEMA enabled the quantification of PR for the first time in such tissues as the bursa and small intestine even of immature animals. Journal of Endocrinology (1991) 129, 189–196

Development of an indirect ELISA assay for detecting antibodies against mammalian reovirus in pigs

2018 ◽  
Vol 262 ◽  
pp. 61-64 ◽  
Author(s):  
Zhijie Li ◽  
Xiaozhan Zhang ◽  
Xiaoliang Hu ◽  
Jin Tian ◽  
Hongtao Kang ◽  
...  
Keyword(s):  
Indirect Elisa ◽  
Elisa Assay

scholarly journals New chromogens for alkaline phosphatase histochemistry: salmon and magenta phosphate are useful for single- and double-label immunohistochemistry.

1994 ◽  
Vol 42 (4) ◽  
pp. 551-554 ◽  
Author(s):  
C Avivi ◽  
O Rosen ◽  
R S Goldstein
Keyword(s):  
Alkaline Phosphatase ◽  
Monoclonal Antibodies ◽  
Molecular Biology ◽  
Horseradish Peroxidase ◽  
Reaction Products ◽  
Immunocytochemical Detection ◽  
Hematoxylin Staining ◽  
Double Label ◽  
Biology Research

Two new substrate chromogens for alkaline phosphatase (ALP) detection have been recently synthesized for use in molecular biology research, salmon and magenta phosphate. We show here that these two chromogens have advantageous characteristics for immunocytochemistry as well. Their relatively delicate pink- and magenta-colored products do not mask the colors produced by other staining procedures. In addition, the reaction products of these substrates are insoluble in water, ethanol, and xylene, permitting the use of regressive hematoxylin staining procedures and coverslipping in permanent resin-based media. Most importantly, when these ALP substrates are used in double-label immunocytochemistry in combination with horseradish peroxidase-diaminobenzidine (HRP-DAB) and counterstained with hematoxylin, all three colors can be easily distinguished. An application using these substrates for simultaneous immunocytochemical detection of two monoclonal antibodies of different classes, in combination with hematoxylin staining, is illustrated.

Heat-Resistant Psychrotrophic Bacteria Isolated from Pasteurized Milk1

1977 ◽  
Vol 40 (2) ◽  
pp. 101-108 ◽  
Author(s):  
C. J. WASHAM ◽  
H. C. OLSON ◽  
E. R. VEDAMUTHU
Keyword(s):  
Thermal Resistance ◽  
Shelf Life ◽  
Detailed Examination ◽  
Genus Bacillus ◽  
Pasteurized Milk ◽  
Psychrotrophic Bacteria ◽  
Milk Samples ◽  
Heat Resistant ◽  
Different Types

Psychrotrophic bacteria were isolated from 227 pasteurized milk samples which had a shelf life in excess of 20 days at 7.2 C. Of 700 cultures isolated, 135 were resistant to heating at 72 C for 16 sec and were able to re-establish growth at 7.2 C. Thirty-five cultures, representing 15 different types were subjected to detailed examination to determine their actions on refrigerated milk, growth temperatures, thermal resistance at various temperatures, and their identities. The spore-forming genus Bacillus occured most frequently. The non-sporing types were assigned to the genera Arthrobacter, Microbacterium, Streptococcus, and Corynebacterium.

scholarly journals Determination of Mycoplasma bovis specific antibodies in blood sera of asymptomatic carriers-calves in three farms in the Republic of Serbia by using indirect ELISA assay

2017 ◽  
Vol 65 (2) ◽  
pp. 79
Author(s):  
D. VOJINOVIĆ ◽  
A. VASIĆ ◽  
J. ŽUTIĆ ◽  
B. DURIČIĆ ◽  
Z. ILIĆ ◽  
...  
Keyword(s):  
Indirect Elisa ◽  
Mycoplasma Bovis ◽  
Elisa Assay ◽  
Serum Samples ◽  
Asymptomatic Carriers ◽  
Elisa Kit ◽  
Cattle Blood ◽  
The Republic ◽  
The Individual

Blood serum samples of asymptomatic carriers-calves were collected from three farms in the territory of the Republic of Serbia during 2011 and 2012. Commercial Mycoplasma bovis ELISA kit (Bio-X Diagnostics, Belgique) for serological diagnosis from cattle blood sera and milk was used in this research. Calves’ blood sera were tested using immunoenzymatic indirect ELISA assay as described by manufacturer’s instructions. From 5603 blood sera of asymptomatic carriers-calves 144 (2,57%) samples were tested positive for the presence of specific Mycoplasma bovis antibodies. In three different farms proportions of seropositive samples varied from 0,32% to 10,6% in regard to total number of tested samples from the individual farms. In this paper we present the results of Mycoplasma bovis prevalence in asymptomatic carriers-calves.

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