• Journal of Veterinary Science
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• v.21 no.4
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• pp.63.1-63.9
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• 2020

Background: Canine adenovirus type 2 (CAV-2) induces infectious laryngotracheitis in members of the family Canidae, including dogs. To date, no ELISA kits specific for CAV-2 antibody have been commercialized for dogs in Korea. Objectives: We aimed to develop new indirect enzyme-linked immunosorbent assay (I-ELISA) to perform rapid, accurate serological surveys of CAV-2 in dog serum samples. Methods: In total, 165 serum samples were collected from dogs residing in Chungbuk and Gyeongbuk provinces between 2016 and 2018. The Korean CAV-2, named the APQA1701-40P strain, was propagated in Madin-Darby canine kidney cells and purified in an anion-exchange chromatography column for use as an antigen for I-ELISA. The virus-neutralizing antibody titers of CAV-2 in the dog sera were measured by virus neutralization (VN) test. Results: We compared the results obtained between the VN and new I-ELISA tests. The sensitivity, specificity, and accuracy of new I-ELISA were 98.6%, 86.4% and 97.0% compared with VN test, respectively. New I-ELISA was significantly correlated with VN (r = 0.91). Conclusions: These results indicate that new I-ELISA is useful for sero-surveillance of CAV-2 in dog serum.

• Journal of Microbiology
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• v.41 no.2
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• pp.148-151
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• 2003

Waterborne parasite infections are considered a reemerging threat. Most studies on the epidemiology of human cryptosporidiosis, giardiasis, and amebiasis have been carried out in developed countries, and there is little data on the occurrence of these infections in other areas. The objective of this study was to investigate the presence of waterborne parasites such as Cryptosporidium parvum, Giardia lamblia and Entamoeba histolytica in various water samples in Ankara, turkey. A total of 85 samples were examined, 43 from the municipal water supply, 34 from wells, 6 from the Ankara River, and 2 from two untreated dams; by conventional microscopy, immunologically and by polymerase chain reaction (PCR). Oocysts of C. parvum and cysts of G. lamblia were detected by using an indirect fluorescence (antigen) assay, whereas an enzyme linked immunosorbent assay was used to detect the cysts of E. histolytica and E. dispar. In addition, PCR was used for E. histolytica, E. dispar, C. parvum and G. lamblia detection. G. lamblia was found in 2 of the 34 well water samples, and parasites were found in 3 of the 6 Ankara River samples. The 1$\^$st/ contained E. histolytica cysts and Strongyloides stercoralis larvae. the 2$\^$nd/ E. histolytica cysts, and Trichuris trichiura eggs, and the 3$\^$rd/ C. parvum oocysts only. No parasite was observed in the municipal water samples and untreated dam water samples. These results extend our knowledge on waterborne parasites, such occurrence information on waterborne pathogens assists the management and treatment of municipal water.

• Journal of Veterinary Science
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• v.23 no.5
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• pp.32.1-32.11
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• 2022

Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases. The stability and sensitivity were evaluated using an accelerated thermostability test. Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. Conclusions: The spbELISA has practical applications in assessing the vaccination status of large pig herds.

• The Korean Journal of Pesticide Science
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• v.5 no.2
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• pp.1-12
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• 2001

A competitive indirect enzyme-linked immunosorbent assay (ci ELISA) for the organophosphorus insecticide acephate, O,S-dimethyl acetylphosphoramidothioate, was developed using a polyclonal antibody. Three different haptens mimicking the analyze and containing hexanoic acid moiety as a linker were synthesized, and then conjugated with the carrier proteins bovine serum albumin and keyhole limpet hemocyanin by the N-hydroxysuccinimide active ester method. Polyclonal antibodies raised against hapten-KLH conjugates in rabbits and the hapten-BSA conjugates as coating antigens were screened and selected for the assay in the homologous and/or heterologous ELISA system. The effects of various assay conditions, including blocking reagents, detergent content, organic solvents, pH, and preincubation of tile mixture of the polyclonal antibody and the analyze on the sensitivity were evaluated. The $IC_{50}$ value of acephate of 110 ng/mL was obtained in an optimized heterologous system using hapten-3-BSA as a coating antigen and a polyclonal antibody 8377, showing the detection range of 10-1000 ng/mL and the lowest detection limit of 4 ng/mL. The cross-reactivities of the structurally related insecticides, including methamidophos were less than 0.02%. These results indicate that the ELISA could be a convenient and alternative tool for monitoring acephate residues in agricultural products and environmental samples.

• Food Science of Animal Resources
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• v.34 no.5
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• pp.570-575
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• 2014

This study was conducted to compare the effects of two forms of radiation (electron and X-ray; generated by an electron beam accelerator) on the conformation and antigenic properties of hen's egg albumin, ovalbumin (OVA), which was used as a model protein. OVA solutions (2.0 mg/mL) were individually irradiated by electron beam or X-ray at the absorbed doses of 0 (control), 2, 4, 6, 8, and 10 kGy. No differences between the two forms of radiation on the structural properties of OVA were shown by spectrometric and electrophoretic analyses. The turbidity of OVA solution increased and the main OVA bands on polyacrylamide gels disappeared after irradiation, regardless of the radiation source. In competitive indirect enzyme-linked immunosorbent assay, OVA samples irradiated by electron beam or X-ray showed different immunological responses in reactions with monoclonal and polyclonal antibodies (immunoglobulin G) produced against non-irradiated OVA. The results indicate that electron beam irradiation and X-ray irradiation produced different patterns of structural changes to the OVA molecule.

• Korean Journal of Veterinary Research
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• v.51 no.4
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• pp.277-280
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• 2011

Brucellosis is the most important bacterial disease of livestock in Bangladesh. The present study was conducted to determine the seroprevalence of brucellosis in goat and sheep in Mymensingh and Bogra districts of Bangladesh using slow agglutination test and Rose Bengal test as screening test and indirect enzyme linked immunosorbent assay as confirmatory test. Questionnaire based data on age, gender, area, client's complaint, number of animals in herds, disease history, reproductive problems such as abnormal uterine discharge, abortion or previous abortion in sheep and goat and their reproductive diseases were recorded. A total of 200 sera samples were collected from 80 sheep and 120 goats. The prevalence of brucellosis in goat was 2.50% and 1.25% in sheep. Positive reactors were only detected in female of both goat and sheep. In this study, there existed a significant association among abortion and the prevalence of brucellosis (p < 0.01). The prevalence of brucellosis in sheep and goat in Bangladesh is not negligible, and it is therefore worth considering the adoption of preventive measures.

• Food Science and Biotechnology
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• v.17 no.5
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• pp.919-924
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• 2008

The aim of this study was to observe the changes in allergenicity of saeujeot (salted and fermented shrimp) using a competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA). The fermentation conditions tested for saeujeot consisted of various temperatures (25, 15, and $5^{\circ}C$) and salt concentrations (25, 15, and 10%). When saeujeot was fermented at a low salt concentration and high temperature, the binding ability of mAb and shrimp-allergic patient serum to allergen was significantly decreased. In particular, the binding ability of mAb to allergen in saeujeot fermented with 10% salt at $25^{\circ}C$ for 5 days decreased to 5%. Also, the binding ability of shrimp-allergic patient serum to allergen in saeujeot fermented for 5 days with 10% salt at $25^{\circ}C$ was 8%. In conclusion, the binding of mAb and shrimp-allergic patient serum to tropomyosin in saeujeot decreased with longer fermentation periods, lower salt concentrations (10%), and higher temperatures ($25^{\circ}C$).

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.7
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• pp.1169-1174
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• 2004

This study was carried out to assess the effects of electron beam (EB) radiation on the conformational changes of ovalbumin (OVA), based on the early works using gamma irradiation. The applied doses of OVA used were 3,5,7, and 10 kGy, respectively. The conformational alterations were measured with SDS-PAGE, GPC-HPLC, and competitive indirect enzyme-linked immunosorbent assay (Ci-ELISA) using monoclonal anti-OVA IgG antibody. Irradiation caused a degradation and/or an aggregation of OVA molecule. Immunochemical structures of irradiated OVA were altered by irradiation. Effects of gamma and electron beam radiation were similar at the same absorbed doses. These results may be used for inhibition of food allergy and development of immunogen with EB radiation.

• Biomedical Science Letters
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• v.1 no.1
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• pp.1-8
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• 1995

The present study was undertaken to determine the dissociation constant (Kd)of monoclonal antibody to human apolipoprotein A-I (apo A-I) using enzyme-linked immunosorbent assay (ELISA). First the monoclonal antibody was incubated in solution with the antigen until the equilibrium was reached; then the free antibody which remains unsaturated at equilibrium was captured by binding to antigen on the microtiter plate and be measured by a classical indirect ELISA. The value of Kd determined from Scatchard plot was 0.625$\times$10^{-9}$ for purified antibody and 0.720$\times$10$^{-9}$ for unpurified antibody. This method was valuable for the measurement of true dissociation constant and found to be simple, reproducible, and accurate.

• Journal of Korean society of Dental Hygiene
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• v.16 no.3
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• pp.463-469
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• 2016

Objectives: The purpose of the study is to investigate the effect of chewing gum containing Prunus mume extract(PME) on the change of saliva ingredients. On the basis of the biological background of molecules and diagnostic indices in the use of saliva, the mastication effect of chewing gum containing PME was demonstrated in terms of secretory IgA concentration and total protein concentration in stimulated saliva. Methods: This study is an experimental research on the use of a research design before and after applying a randomized control group. Participants were distributed randomly to the experimental group and the control group, respectively. The experiment group was instructed to masticate the chewing gum containing PME for 10 minutes for one month after each meal within 30 minutes. Salivary secretion was collected by the participants between 8 and 10 a.m in the morning in the research office. For the measurement of secretory IgA and total protein concentrations in the saliva, indirect enzyme-linked immunosorbent assay(ELISA) was used. Results: The salivation stimulation rate was significantly increased after four weeks of masticating chewing gum containing PME after each meal(p<0.001). Mastication of chewing gum containing PME for four weeks decreased the concentration of secretory IgA much more significantly than that after mastication for one week(p=0.003). The concentration of total protein in the saliva was decreased after four weeks in the experimental and control groups. Conclusions: Mastication of chewing gum containing PME stimulated salivary secretion and led to oral disease prevention in patients with xerostomia. Furthermore, it seems to be urgent to seek measures that can be utilized in intervention for patients with xerostomia.