Evaluation of Monoclonal Antibody (IgG2bMAb) for Detection of Coproantigen from Experimentally Infected Rats with Strongyloides ratti

Background and Aim: Highly sensitive and specific diagnostic assay for the detection of Strongyloides is needed due to the intermittent and low concentration of eggs, larvae and adult worms that can be found in a faecal specimen. In some cases, repeated sampling of the faecal specimen is required to obtain satisfactory and reliable results. The aim of the study is to develop and evaluates monoclonal antibody-based Sandwich ELISA for the detection of coproantigen associated with Strongyloides infection using S. ratti as a model. Place and Duration of Study: Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, University Malaysia, Between September 2018 and March 2019. Methodology: The monoclonal antibody was raised against a soluble antigen of the infective filariform larvae (iL3) of S. ratti. The monoclonal antibody produced (IgG2bMAb) was evaluated for cross-reactivity against homologous and heterologous helminth antigens such as excretory-secretory (ES), infective larvae (iL3) and coproantigen of S. ratti, adult worms of A. caninum, A. suum, T. canis and T. cati. Results: An IgG2bMAb was observed to react with 30 kDa proteins associated with all homologous antigen from iL3, ES and coproantigen of S. ratti and cross-reacted with one heterologous antigen from adult worm of A. caninum at the same molecular weight. There was no cross-reaction observed with other heterologous antigens from adult worms of T. canis, T. cati and A. suum. The sensitivity of IgG2bMAb for the detection of S. ratti was 85% in Sandwich ELISA. Cross- reaction was observed with hookworm antigen that caused by A. caninum in Western immunoblotting. Conclusion: The results indicated that IgG2b have an immunodiagnostic property as IgG2bMAb and was able to detect antigens from coproantigen related to S. ratti with 85% sensitivity based on Sandwich ELISA) even though cross-reaction was observed with A. caninum. These findings will be very useful to tackle many cases of multiple worms’ infections such as both strongyloidiasis and hookworm. Therefore, we recommend that further evaluation and study in the human area where multiple infections can be common should be carried out.

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Characterization of G12 Sandwich ELISA, a Next-Generation Immunoassay for Gluten Toxicity

Journal of AOAC International ◽

10.5740/jaoacint.sge_halbmayr-jech ◽

2012 ◽

Vol 95 (2) ◽

pp. 372-376 ◽

Cited By ~ 17

Author(s):

Elisabeth Halbmayr-Jech ◽

Elisabeth Hammer ◽

Richard Fielder ◽

Jacqueline Coutts ◽

Adrian Rogers ◽

...

Keyword(s):

Monoclonal Antibody ◽

Celiac Disease ◽

Working Group ◽

Sandwich Elisa ◽

Extraction Methods ◽

Cross Reactivity ◽

Next Generation ◽

Preliminary Results ◽

Sample Extraction

Abstract In this work, a monoclonal antibody called G12, raised against the most immunotoxic peptide to celiac disease patients, was used to develop a sandwich ELISA. Preliminary results on cross-reactivities, recoveries, and extraction methods of the new assay are presented. The assay calibration was performed using material from the Prolamin Working Group. The antibody's specificity was determined by cross-reactivity studies on different grains, nuts, oils, and starches. Recovery of the assay was determined by spiking experiments on common food matrixes, as well as on problematic matrixes. Furthermore, sample extraction methods using ethanol, cocktail solution, and a proprietary buffer have been compared.

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Cross-reaction between mouse and rat immunoglobulin G: does it matter in sandwich ELISA?

Journal of Genetic Engineering and Biotechnology ◽

10.1186/s43141-021-00222-2 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Rola Nadeem ◽

Ahmed B. Barakat ◽

Mahmoud M. Bahgat

Keyword(s):

Sandwich Elisa ◽

Cross Reactivity ◽

Antigen Detection ◽

Cross Reaction ◽

Laboratory Animals ◽

Elisa Assay ◽

Swiss Albino Mice ◽

Specific Antibodies ◽

Specific Reaction ◽

Albino Mice

Abstract Background Sandwich ELISA is an ideal antigen detection and quantification assay. Recently, it was used as the basic concept for high technology diagnostics. The specificity of the assay depends on the exclusion of detection cross-reactivity which arises from using two antibodies developed in different species. Since mice and rats are the common laboratory animals used to develop antigen specific antibodies. Therefore, the questions we addressed here were (1) can one use antigen-specific antibodies raised in mice and rats in the same assay to specifically detect/quantify antigens? and (2) which antibodies of the two rodents should be placed for capturing and for detection in the antigen-detection sandwich? Results Direct ELISA assay was used to assess for the specific reaction of the HRP-conjugated antibody to the target serum. First reaction was to compare between either conjugate anti-rat IgG (homologous) or anti-mouse IgG (heterologous) for the detection of rat sera IgG. Following the dilution factor optimization, the O.D. were 0.744±0.051 and 0.604±0.05, respectively (p= .004). The difference in mean O.D. of 0.14 reflected an unaccepted non-specific reaction. The second reaction was to compare between either conjugate anti-mouse IgG (homologous) and anti-rat IgG (heterologous) for the detection of mouse sera IgG. The recorded O.D. were 0.9414±0.14 and 0.317 ±0.141, respectively (p= .0002). The improved difference in mean O.D. of 0.624 reflecting a minimized cross-reaction. Conclusions Our results suggest that it is possible to use both Swiss albino mice and albino rats in a single sandwich ELISA, given that the captured antibody species to be from the Swiss albino mice and the detection antibody to be from the albino rat. The described working details are limited to the source of the antibodies used in the study. However, the approach stresses on the importance of such optimization steps before making any interpretations based on the antigen detection. To our knowledge, this study is the first to cover the optimal order of the capturing and the detection antibodies in a sandwich ELISA assay. In addition to addressing the possible interfering cross-reactivity that result from using mouse and rat serum antibodies in a single assay. Graphical abstract

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Serological responses in sheep injected with plasmids encoding bovine herpesvirus 1 (BHV-1) gD glycoprotein

Arquivo Brasileiro de Medicina Veterinária e Zootecnia ◽

10.1590/s0102-09352003000300002 ◽

2003 ◽

Vol 55 (3) ◽

pp. 256-261

Author(s):

A.L. Cândido ◽

M. Resende ◽

L.R.G. Bessa ◽

R.C. Leite

Keyword(s):

Bovine Herpesvirus ◽

Cross Reactivity ◽

Glycoprotein D ◽

Serum Antibodies ◽

Bovine Herpesvirus 1 ◽

Heterologous Antigen ◽

Specific Antibodies ◽

Genetic Vaccine ◽

Homologous Antigen ◽

Herpesvirus 1

A genetic vaccine consisting of the bovine herpesvirus-1.2a (BHV-1.2a) glycoprotein D (gD) gene under the control of the cytomegalovirus immediate-early promoter/enhancer was generated and administered to sheep intramuscularly in the neck. All animals developed serum antibodies which recognized the homologous antigen (BHV-1.2a strain BH-83) and also exhibited cross-reactivity against the heterologous antigen (BHV-5 strain EVI-190). Three intramuscularly injections were given but serological responses were not improved after the second inoculation. Specific antibodies were detected against BHV-1.2a until at least 12 months after the first inoculation. However, the capacity to induce antibodies against BHV-5 was lower and of shorter duration than to BHV-1.2a.

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Production of a monoclonal antibody specific for ovine immunoglobulin E and its application to monitor serum IgE responses to Haemonchus contortus infection

Parasitology ◽

10.1017/s0031182096008633 ◽

1997 ◽

Vol 114 (4) ◽

pp. 395-406 ◽

Cited By ~ 56

Author(s):

F. N. J. KOOYMAN ◽

P. J. S. VAN KOOTEN ◽

J. F. HUNTLEY ◽

A. MacKELLAR ◽

A. W. C. A. CORNELISSEN ◽

...

Keyword(s):

Monoclonal Antibody ◽

Haemonchus Contortus ◽

Immunoglobulin E ◽

Polyclonal Antibodies ◽

Sandwich Elisa ◽

Specific Ige ◽

Cross Reactivity ◽

Total Serum ◽

Western Blots ◽

Serum Ige

Part of the Cε3–Cε4 region of the ovine immunoglobulin E (IgE) gene (nucleotides 1111–1575) was amplified by PCR. The recombinant protein (recIgE1-2) was expressed in E. coli and both monoclonal and polyclonal antibodies were produced. These antibodies recognized recIgE1-2 and native IgE on Western blots and in ELISA. The polyclonal serum showed cross-reactivity with other sheep immunoglobulin classes. The monoclonal antibody was specific for ovine IgE and goat IgE. Infection of sheep with the abomasal nematode Haemonchus contortus resulted in elevated IgE levels in serum 2–4 weeks after infection, as measured by sandwich ELISA using the rabbit polyclonal as capture antibody and the monoclonal antibody against ovine IgE as second antibody. A negative correlation between worm counts and total serum IgE levels at the end of the experiment was found in repeatedly infected sheep. Significant increased levels of excretory–secretory antigens specific IgE levels were found after H. contortus infection. In contrast, no significant changes in 3rd-stage larvae (L3) antigen-specific IgE titre in sera could be detected after infection.

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Evidence of Cross-Reactivity of ‘Carcinoma-Specific’ KC4 Monoclonal Antibody with Activated Mesothelial Cells and Phytohemagglutinin-Stimulated Lymphocytes

Oncology ◽

10.1159/000226646 ◽

1988 ◽

Vol 45 (5) ◽

pp. 380-383

Author(s):

A. Neubauer ◽

R. Musch ◽

U. Thalmann ◽

H. Grosser ◽

J. Laser ◽

...

Keyword(s):

Monoclonal Antibody ◽

Cross Reactivity ◽

Mesothelial Cells

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Do intestinal parasites interfere with the seroepidemiologic surveillance of Schistosoma mansoni infection?

Epidemiology and Infection ◽

10.1017/s095026880005264x ◽

1996 ◽

Vol 116 (3) ◽

pp. 323-329 ◽

Cited By ~ 14

Author(s):

B. Alarcón De Noya ◽

C. Colmenares ◽

S. Losada ◽

Z. Fermin ◽

G. Masroua ◽

...

Keyword(s):

Schistosoma Mansoni ◽

False Positive ◽

Total Population ◽

Intestinal Parasites ◽

Field Work ◽

Cross Reactivity ◽

Cross Reaction ◽

Protein Fractions ◽

Egg Antigen ◽

Immunoenzymatic Assay

SUMMARYIn view of the known cross-reactivity of sera from patients with intestinal parasites to some Schistosoma mansoni antigens, field work was conducted in an area of Venezuela non-endemic for schistosomiasis using the routine immunoenzymatic assay (ELISA) with soluble egg antigen (SEA). False positive reactions represented 15·3% of the total population as determined by SEA–ELISA. SEA-immunoblotting of the false positive sera indicated that protein fractions of 91 and 80 kDa appear to be responsible for cross-reactivity. Sera from hookworm infected individuals produced a higher frequency and intensity of cross-reaction than other sera. SEA-fractions of 105, 54, 46, 42, 32, 25 and 15 kDa were the most specific.

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Trypanosoma cruzi: identification of specific epimastigote antigens by human immune sera

Memórias do Instituto Oswaldo Cruz ◽

10.1590/s0074-02761989000300004 ◽

1989 ◽

Vol 84 (3) ◽

pp. 309-314 ◽

Cited By ~ 5

Author(s):

M. G. Morgado ◽

J. Ivo-dos-Santos ◽

R. T. Pinho ◽

E. Argüelles ◽

J. M. Rezende ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Trypanosoma Cruzi ◽

Western Blot ◽

Chagas Disease ◽

Cross Reactivity ◽

Cross Reaction ◽

The Other ◽

Molecular Weights ◽

Human Visceral Leishmaniasis ◽

Human Immune

Soluble antigens from epimastigotes of Trypanosoma cruzi were analyzed by western blot in terms of their reactivity with sera from patients with Chagas' disease. In addition, sera from patients with visceral (AVL) and tegumentar leishmaniasis (ATL) were also tested in order to identify cross-reactivities with Trypanosoma cruzy antigens. Twenty eight polypeptides with molecular weights ranging from 14 kDa to 113 kDa were identified with sera from Chagas' disease patients. An extensive cross-reactivity was observed when sera from human visceral leishmaniasis were used, while only a slight cross-reaction was observed with sera from tegumentar leishmaniasis. On the other hand, 10 polypeptidesspecifically reacting with sera from Chagas' disease patients were identified. Among them, the antigens with molecular weights of 46 kDa and 25 kDa reacted with all sera teste and may be good candidates for specific immunodiagnosis of Chagas' disease.

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Lactate Dehydrogenase Monoclonal Antibody Sandwich ELISA To Determine Cooking Temperature of Ground Beef

Journal of Agricultural and Food Chemistry ◽

10.1021/jf960327c ◽

1996 ◽

Vol 44 (12) ◽

pp. 4048-4051 ◽

Cited By ~ 7

Author(s):

A. Orta-Ramirez ◽

C. H. Wang ◽

M. M. Abouzied ◽

G. J. Veeramuthu ◽

J. F. Price ◽

...

Keyword(s):

Monoclonal Antibody ◽

Lactate Dehydrogenase ◽

Sandwich Elisa ◽

Ground Beef ◽

Cooking Temperature

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EVALUATION OF ELEVEN IMMUNOCHROMATOGRAPHIC ASSAYS FOR SARS-CoV-2 DETECTION: INVESTIGATING DENGUE CROSS-REACTION

10.1101/2020.10.09.20210039 ◽

2020 ◽

Author(s):

Beatriz Araujo Oliveira ◽

Lea Campos de Oliveira ◽

Franciane Mendes de Oliveira ◽

Geovana Maria Pereira ◽

Regina Maia de Souza ◽

...

Keyword(s):

Sensitivity And Specificity ◽

High Sensitivity ◽

Diagnostic Techniques ◽

Cross Reactivity ◽

Cross Reaction ◽

List Type ◽

Rt Pcr ◽

Serum Samples ◽

Igm Antibodies ◽

Good Agreement

AbstractBackgroundCOVID-19 disease (Coronavirus disease 2019) caused by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2) is widespread worldwide, affecting more than 11 million people globally (July 6th, 2020). Diagnostic techniques have been studied in order to contain the pandemic. Immunochromatographic (IC) assays are feasible and low cost alternative for monitoring the spread of COVID-19 in the population.MethodsHere we evaluate the sensitivity and specificity of eleven different immunochromatographic tests in 98 serum samples from confirmed cases of COVID-19 through RT-PCR and 100 negative serum samples from blood donors collected in February 2019. Considering the endemic situation of Dengue in Brazil, we also evaluated the cross-reactivity with Dengue using 20 serum samples from patients with confirmed diagnosis for Dengue collected in early 2019 through four different tests.ResultsOur results demonstrated agreement between immunochromatographic assays and RT-PCR, especially after 10 days since the onset of symptoms. The evaluation of IgG and IgM antibodies combined demonstrated a strong level of agreement (0.85) of IC assays and RT-PCR. It was observed cross-reactivity between Dengue and COVID-19 using four different IC assays for COVID-19 diagnosis. The specificity of IC assays to detected COVID-19 IgM antibodies using Dengue serum samples varied from 80% to 85%; the specificity of IgG detection was 100% and total antibody was 95%.ConclusionsWe found high sensitivity, specificity and good agreement of IC assays, especially after 10 days onset of symptoms. However, we detected cross-reactivity between Dengue and COVID-19 mainly with IgM antibodies demonstrating the need for better studies about diagnostic techniques for these diseases.HighlightsImmunochromatographic assays demonstrated high sensitivity and specificity and good agreement with the gold-standard RT-PCR;Increase in sensitivity and specificity of assays using samples collected after the 10th day of symptoms;Cross-reaction with Dengue serology in evaluation of IgM.

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