Detection of American foulbrood disease of the honeybee, using a monoclonal antibody specific to Bacillus larvae in an enzyme-linked immunosorbent assay

1990 ◽  
Vol 36 (10) ◽  
pp. 732-735 ◽  
Author(s):  
Perry E. Olsen ◽  
Gordon A. Grant ◽  
Donald L. Nelson ◽  
Wendell A. Rice
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
The United States ◽  
Bacillus Species ◽  
Enzyme Linked Immunosorbent Assay ◽  
American Foulbrood ◽  
Field Samples ◽  
Elisa Technique ◽  
American Foulbrood Disease

American foulbrood, a virulent disease of honeybees, is caused by Bacillus larvae. A murine hybridoma which secretes an IgM monoclonal antibody reactive with B. larvae was derived and an indirect enzyme-linked immunosorbent assay (ELISA) using this monoclonal antibody was evaluated for use in detection of the pathogen. The ELISA detected B. larvae strains from Alberta. and British Columbia, Canada, and from the United States but did not detect six other Bacillus species. The ELISA also detected the presence of B. larvae in field samples collected from commercial apiaries. Results of testing against cultures of B. larvae and infected bee larvae from hives indicate that the monoclonal antibody has, in conjunction with the indirect ELISA technique, potential for use in the diagnosis and confirmation of American foulbrood in commercial apiaries and in honeybee pathology research. Key words: American foulbrood, Bacillus larvae, honeybee disease.

Use of a Monoclonal Antibody and Two Enzyme-Linked Immunosorbent Assay Formats for Detection and Quantification of the Substitution of Caprine Milk for Ovine Milk

1997 ◽  
Vol 60 (8) ◽  
pp. 973-977 ◽  
Author(s):  
ANA I. HAZA ◽  
PALOMA MORALES ◽  
ROSARIO MARTÍIN ◽  
TERESA GARCÍIA ◽  
GONZALO ANGUITA ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Goat’S Milk ◽  
Elisa Format ◽  
Ewe's Milk ◽  
Goat's Milk ◽  
Detection And Quantification

A stable hybridoma cell line (B2B) has been produced that secretes a monoclonal antibody (MAb) specific for goat's milk αS2-casein. The MAb B2B was used in two enzyme-linked immunosorbent assay (ELISA) formats for the detection and quantification of the presence of goat's milk in ewe's milk. In the indirect ELISA format the limit of detection was 0.5 to 15% (vol/vol) substitution of goat's milk for ewe's milk. Afterwards, a competitive indirect ELISA was successfully developed for the detection of 0.25 to 15% (vol/vol) of goat's milk in ewe's milk. This competitive indirect ELISA is a very sensitive assay; it can be performed in less than 5 h and is not influenced by the heat treatment of milk.

scholarly journals Validation of a Monoclonal Antibody-Based Capture Enzyme-Linked Immunosorbent Assay for Detection of Campylobacter fetus

2005 ◽  
Vol 12 (11) ◽  
pp. 1261-1268 ◽  
Author(s):  
J. Devenish ◽  
B. Brooks ◽  
K. Perry ◽  
D. Milnes ◽  
T. Burke ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Area Under The Curve ◽  
Stomach Contents ◽  
Enzyme Linked Immunosorbent Assay ◽  
Characteristic Analysis ◽  
Serotype A ◽  
Campylobacter Fetus ◽  
Field Samples ◽  
Serotype B

ABSTRACT A monoclonal antibody (MAb)-based antigen capture enzyme-linked immunosorbent assay (ELISA) was compared with the routine culture methodology for the detection of Campylobacter fetus subspecies from bovine and ovine field samples inoculated into Clark's transport enrichment medium (TEM). The work was a collaboration between two different diagnostic laboratories, one in Canada and the other in England. In both labs, TEM samples were incubated for 4 days at 35°C and then tested by culture and ELISA. The ELISA consisted of initial screening with MAb M1825 against C. fetus subspecies core lipopolysaccharide (LPS). All samples positive on ELISA screening were then retested by ELISA with MAb M1825 and MAbs M1177, M1183, and M1194, which recognize serotype A- and/or serotype B-specific C. fetus subspecies LPS epitopes. The Canadian samples consisted of 1,060 preputial washings from 529 bulls, of which 18 were positive by both culture and ELISA and 1,042 were negative by both methods. The English samples consisted of 321 tissue specimens, mostly stomach contents and placentas, from 190 aborted ovine and bovine fetuses. A total of 262 samples were negative by culture and ELISA, 52 samples were positive by culture and ELISA, and 7 samples were culture negative but ELISA positive. The results for all 70 culture-positive isolates were confirmed by conventional biochemical methods as C. fetus subsp. fetus, with 39 presumptively identified by the ELISA as serotype A and 30 presumptively identified as serotype B and with one sample containing isolates presumptively identified as serotype A and serotype B. A receiver operating characteristic analysis of the combined ELISA data from both countries resulted in an area under the curve of 0.997, with a sensitivity of 100% and a specificity of 99.5% relative to the results of culture. The data confirm that this ELISA method can be used as an excellent test for the screening of field samples in TEM for the presence of C. fetus subspecies.

scholarly journals Development of a triple antibody sandwich enzyme-linked immunosorbent assay for cassava mosaic disease detection using a monoclonal antibody to Sri Lankan cassava mosaic virus

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Saengsoon Charoenvilaisiri ◽  
Channarong Seepiban ◽  
Mallika Kumpoosiri ◽  
Sombat Rukpratanporn ◽  
Nuchnard Warin ◽  
...  
Keyword(s):  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Mosaic Disease ◽  
Cassava Mosaic Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sri Lankan ◽  
Stem Cuttings ◽  
Field Samples ◽  
Cassava Stem ◽  
Cassava Mosaic Virus

Abstract Background Cassava mosaic disease (CMD) is one of the most devastating viral diseases for cassava production in Africa and Asia. Accurate yet affordable diagnostics are one of the fundamental tools supporting successful CMD management, especially in developing countries. This study aimed to develop an antibody-based immunoassay for the detection of Sri Lankan cassava mosaic virus (SLCMV), the only cassava mosaic begomovirus currently causing CMD outbreaks in Southeast Asia (SEA). Methods Monoclonal antibodies (MAbs) against the recombinant coat protein of SLCMV were generated using hybridoma technology. MAbs were characterized and used to develop a triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) for SLCMV detection in cassava leaves and stems. Assay specificity, sensitivity and efficiency for SLCMV detection was investigated and compared to those of a commercial ELISA test kit and PCR, the gold standard. Results A TAS-ELISA for SLCMV detection was successfully developed using the newly established MAb 29B3 and an in-house polyclonal antibody (PAb) against begomoviruses, PAb PK. The assay was able to detect SLCMV in leaves, green bark from cassava stem tips, and young leaf sprouts from stem cuttings of SLCMV-infected cassava plants without cross-reactivity to those derived from healthy cassava controls. Sensitivity comparison using serial dilutions of SLCMV-infected cassava sap extracts revealed that the assay was 256-fold more sensitive than a commercial TAS-ELISA kit and 64-fold less sensitive than PCR using previously published SLCMV-specific primers. In terms of DNA content, our assay demonstrated a limit of detection of 2.21 to 4.08 × 106 virus copies as determined by quantitative real-time PCR (qPCR). When applied to field samples (n = 490), the TAS-ELISA showed high accuracy (99.6%), specificity (100%), and sensitivity (98.2%) relative to the results obtained by the reference PCR. SLCMV infecting chaya (Cnidoscolus aconitifolius) and coral plant (Jatropha multifida) was also reported for the first time in SEA. Conclusions Our findings suggest that the TAS-ELISA for SLCMV detection developed in this study can serve as an attractive tool for efficient, inexpensive and high-throughput detection of SLCMV and can be applied to CMD screening of cassava stem cuttings, large-scale surveillance, and screening for resistance.

scholarly journals Unreliable Measurement of Basophil Maximum Leukotriene Release with the Bühlmann CAST 2000 Enzyme-Linked Immunosorbent Assay Kit

2006 ◽  
Vol 13 (3) ◽  
pp. 420-422 ◽  
Author(s):  
S. E. Burastero ◽  
C. Paolucci ◽  
D. Breda ◽  
G. Monasterolo ◽  
R. E. Rossi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
False Positive ◽  
Positive Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Positive Results

ABSTRACT The Bühlmann CAST 2000 enzyme-linked immunosorbent assay is a potentially useful assay for measuring sulfidoleukotrienes released in vitro by allergen-challenged basophils. However, we observed that the positive-control reagent yielded positive signals in cell-free systems. These false-positive results depended on using a mouse anti-FcεRI monoclonal antibody and were prevented by degranulation-inducing reagents other than mouse monoclonal antibodies.

Use of a monoclonal antibody to estrone-3-glucuronide in an enzyme-linked immunosorbent assay (ELISA)

1990 ◽  
Vol 36 (5) ◽  
pp. 439-443 ◽  
Author(s):  
Peter A. Elder ◽  
Laurette Manley ◽  
John G. Lewis
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay

Development of an Enzyme-Linked Immunosorbent Assay and Immunoaffinity Column Chromatography for Saikosaponin d Using an Anti-Saikosaponin d Monoclonal Antibody

Planta Medica ◽  
2016 ◽  
Vol 82 (05) ◽  
pp. 432-439 ◽  
Author(s):  
Jiayang Sai ◽  
Yan Zhao ◽  
Wenchao Shan ◽  
Baoping Qu ◽  
Yue Zhang ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Column Chromatography ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunoaffinity Column

Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for the analysis of the new fungicide 2-allylphenol in strawberry fruits

2010 ◽  
Vol 120 (4) ◽  
pp. 1178-1184 ◽  
Author(s):  
Yuanyuan Xia ◽  
Qing X. Li ◽  
Shuangjun Gong ◽  
Yong Li ◽  
Yongsong Cao ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay

Evaluation of a monoclonal antibody-based enzyme-linked immunosorbent assay for detection of Campylobacter fetus in bovine preputial washing and vaginal mucus samples

2004 ◽  
Vol 103 (1-2) ◽  
pp. 77-84 ◽  
Author(s):  
B.W. Brooks ◽  
J. Devenish ◽  
C.L. Lutze-Wallace ◽  
D. Milnes ◽  
R.H. Robertson ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Campylobacter Fetus

scholarly journals Evaluation of a monoclonal antibody affinity purified antigen for zymodeme specific serological diagnosis of Trypanosoma cruzi infection

1987 ◽  
Vol 82 (1) ◽  
pp. 81-85 ◽  
Author(s):  
Mauro Schechter
Keyword(s):  
Monoclonal Antibody ◽  
Trypanosoma Cruzi ◽  
Affinity Chromatography ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Monoclonal Antibody ◽  
Antibody Affinity ◽  
Specific Diagnosis ◽  
Purified Antigen ◽  
Cruzi Infection

Theoretically, serological assays with affinity purified marker antigens can allow strain-specific diagnosis even when parasites cannot be retrieved from and infected host. A Trypanosoma cruzi antigen was purified by affinity chromatography using a zymodeme (Z) 2 specific monoclonal antibody (2E2C11). An indirect enzyme-linked immunosorbent assay (ELISA) based on the purified antigen could discriminate between sera from rabbits immunized with T. cruzi zymodeme clones but could not discriminate between sera from mice infected with different zymodemes.

Sign in / Sign up

Export Citation Format

Share Document