• Korean Journal of Veterinary Service
• /
• v.26 no.3
• /
• pp.215-219
• /
• 2003

This study was conducted to investigate the similarity between hemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay(ELISA) titers and sample to positive ratio (S/P ratio) of Newcastle disease(ND) virus. To perform this study, the 372 sera of broiler chicks and 120 sera of layers and breed chicks were collected from slaughter house and farms, respectively. As a result of HI test out of different chicks, the positive percentage of ND antibody titer of broiler, layer and breeder, when a standard positive HI titer were '2', was 84.4%, 100% and 100%, respectively. The positive percentage of ND antibody titer by ELISA was shown 38.4%, 100% and 100% and S/P ratio were also shown 81.5%, 98.2% and 99.2%, respectively. The results of comparative survey with same sera by two experimental methods were as follows; In low HI titer, ELISA titer was not similar to HI titer, but S/P ratio was similar to it. In high HI titer, ELISA titer was not similar to HI titer. Therefore, HI titer was more similar to S/P ratio than ELISA titer.

• BMB Reports
• /
• v.30 no.5
• /
• pp.326-331
• /
• 1997

To construct a competitive ELISA standard curve for the detection of glucose-6-phosphate debydrogenase (G6PD), we used highly purified native G6PD (nG6PD) as both immobilized and soluble antigens and anti-G6PD serum raised against nG6PD as antibody. The polystyrene cuvettes coated with nG6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of nG6PD as competitors followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA did not exhibit the typical sigmoidal dose-response curve characteristic of competition immunoassays under the optimal concentrations of antigen and antibody. The soluble nG6PD used as competitor failed to effectively inhibit the binding of antibodies to the immobilized nG6PD. The addition of NADP, a cofactor of G6PD enzyme, to coating buffer used for immobilizing nG6PD to the cuvettes and PBS-Tween-BSA buffer for diluting competitors did not improve the inhibition of antibody binding to immobilized nG6PD by soluble n/G6PD. The addition of BSA to coating buffer did not increase inhibition, either. Surprisingly, when partially active G6PD (paG6PD), obtained by repeated freeze-thawing, was used as competitor, the antibody binding to either immobilized nG6PD or immobilized paG6PD was inhibited 49-58%. We conclude that an effective competitive ELISA system with nG6PD enzyme and anti-G6PD serum for the detection of G6PD may not be established due to the poor inhibition of antibody binding to immobilized nG6PD by soluble nG6PD under the present assay conditions and that the inhibition may be improved by using an inactivated enzyme as competitor regardless of the type of immobilized antigen used. These results imply that the immobilized nG6PD may undergo denaturation upon binding to the polystyrene cuvettes and that our anti-G6PD serum may recognize denatured enzyme better than active enzyme.

• Parasites, Hosts and Diseases
• /
• v.29 no.3
• /
• pp.293-310
• /
• 1991

Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.

• Journal of fish pathology
• /
• v.9 no.2
• /
• pp.169-176
• /
• 1996

An indirect double antibody enzyme-linked immunosorbent assay (ELISA) was developed for rapid detection of a new virus isolated from abnormally swimming salmonid fish, RVS (Retrovirus of salmonid). Results using brain tissue homogenates, and infected cell cultures are described. The sensitivity of the methods is $10^{2.6}$ $TCID_{50}/100{\mu}l$ of the examined cell culture fluid. The specificity was confirmed by the ELISA inhibition test and virological examinations. Viral antigen could be detected in artificially infected fish tissue homogenates. The assay will allow the diagnosis of RVS-infected fish within a day.

• Korean Journal of Veterinary Service
• /
• v.24 no.1
• /
• pp.21-29
• /
• 2001

To compare of serum antibody titers using ELISA and HI, serum samples were collected from 100 breeders and their progeny 550 broilers. The breeders and broilers were vaccinated with infectious bronchitis(IB)- and Newcastle disease(ND)-viruses according to general vaccination program. The antibodies in serum samples against IB and ND viruses were detected by enzyme-linked immunosorbent assay(ELISA) using commercial ELISA kit and hemagglutination inhibition(HI) test. Geometric mean titer(GMT) of ELISA and In titers were monitored from 1-day-old to 35-day-old broilers and compared to those of breeder chickens. The antibody titers of breeders vaccinated with ]B virus showed 47,800, ELISA and 7.2, HI, respectively. Progeny chicks, 1-day-old, vaccinated with IBV showed high antibody titers than those of breed chickens. Those chicks were maintained protective antibody levels until 11-day-old. From 14-day-old, the antibody level decreased below protective levels. In ND, breeders serum antibody titers ELISA and Eiu were 30,200 GMT and 8.7 HI titer, respectively. On 1-day-old chicks, antibody levels was decreased to half in ELISA(16,270) compared with those of breeders, but In titers was 7.4. Progeny broilers, protective antibody level was maintained until 14- day-old by ELISA, but at 11-day-old by HI titers. After then, ND antibody titer was continuously decreased underdefense level. These result indicated that the ELISA method be more sensitive than HI titration to detect serum antibody level for IBV and NDV.

• Korean Journal of Veterinary Research
• /
• v.46 no.3
• /
• pp.185-196
• /
• 2006

Newcastle disease (ND) is a highly contagious disease of poultry that can cause severe economic losses throughout the world. Vaccination has been used for a long time and proved as one of the most effective method to reduce the economic loss due to ND virus infection, The measurement of antibody titer such as hamagglutination-inhibition (Hl) test with sera has been used as a useful method to evaluate the immunity leve of host. However, Hl test is gradually being replaced by the enzyme linked-immunosorbent assay (ELISA), To evaluate the efficacy of ELISA in the chickens vaccinated with different procedure, present study has been performed. After SPF chicks and commercial broilers were vaccinated with different kinds of live vaccines such as V4, VG/GA and/or Bl at various time, the antibody level has been measured using both HI test and ELISA. Challenge test with velogenic viscerotropic NDV was also performed to measure the protective level of antibody. In the SPF chickens, the mean ELISA titer after vaccination and survival rate after challenge was increased and correlated with days post inoculation. More than 80% of chickens with higher than 1,000 ELISA titer after vaccination were survived after challenge with velogenic ND virus and had good correlation between survival rate and antibody titier. In commercial broiler chickens, most of them at market age had low level of ELISA titer regardless of the number of vaccination, and had a low correlation between survival rate and ELISA titer. However, the ELISA titer of remaining birds after challenge was increased. This result indicated that ELISA titer had good response against velogenic NOV infection compared to Hl titer.

• Journal of Food Hygiene and Safety
• /
• v.10 no.4
• /
• pp.213-217
• /
• 1995

A screening method has been developed for detecting sulfamethazine(SMZ) contamination of meat or feeds by using horseradish peroxidase (HRP) labeled protein A (Prot AHRP)and an indirect competitve enzyme-linked immunosorbent assay(ELISA). The assay is based on competitve binding of guinea pig anti-SMZ with SMZ in smaple and SMZ-gelatin conjugate(SMZ.GEL). Percent binding (B.Bo$\times$100) was calculated from the absorbance in the absence (B0) and presence (B) of SMZ. By the sandard curve prepared by plotting log(SMZ) vs percent binding of each known reference solution, the detection limit was 1.0ppb or less. Cross reacton with sulfadimethoxine, sulfaguaniding, sulfamerazine, sulfamthoxpyridazine, sulfanilamide, sulfisomidine and sufisoxazole were not observed. But sulfamerazine crossreacted in the test. The EC-50 value (concentration causing 50% inhibition of color development compared with blank) of sulfamerazine was 2.0 ppm. Further quality control will make the ELISA system ideal for the detection of SMZ in meat or feeds.

• The Journal of the Korean Society for Microbiology
• /
• v.21 no.4
• /
• pp.473-480
• /
• 1986

Mumps is an extremly common infectious disease affecting predominantly young children hut it is not a severe disease in terms of mortality. One hundred and two sera from infants of 3 different groups which are vaccinated, unvaccinated and unknown were detected to mumps antibody. The tests used were Complement Fixation(CF) test, Single Radial Hemolysis(SRH) test, Hemagglutination Inhibition(HI) test, Enzyme Linked Immunosorbent Immunoglobulin G(ELISA IgG) test, Enzyme Linked Immunosorbent Immunoglobulin M(ELISA IgM) test. 1. The rate of positivity for mumps antibody in 102 sera wera 89.16%(74/83) by Hl test, 68.83%(53/77) by ELISA IgG test, 64.58%(62/96) by SRH test, 63.24%(43/68) by ELISA IgM test and 50.00%(49/98) by CF test. 2. The rate of positivity by 5 tests for 55 sera turned out to be very similar with above results respectively. 3. The correlation coefficients(r) between ELISA IgG test ant H1 test, ELISA IgG test and ELISA IgM test were 0.34(P<0.0l) and 0.31(P<0.02), respectively. 4. The percentage of apparently natural infection of mumps seemed to be 65.15%(43/66) in infants. 5. Seroconversion rate of mumps by vaccination were 90.91%(10/11). 6. Among the 53 infants who were tested with ELISA IgG 15 were below 15 months age of(28.30%) and this percentage may be taken as a suggestion that mumps vaccination should be given earlier than present practice. 7. ELISA IgG test was found very sensitive and recommendable method for large scale screening for the presence of antibody to mumps.

• Microbiology and Biotechnology Letters
• /
• v.46 no.3
• /
• pp.269-276
• /
• 2018

Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

• YAKHAK HOEJI
• /
• v.41 no.1
• /
• pp.74-80
• /
• 1997

A polygonal antibody against recombinant hirudin was raised for the development of a ELISA in biological fluids. Recombinant hirudin was conjugated to maleimide activated carrie r protein, KLH and injected to a rabbit. The third booster collection of antiserum was used as primary antibody for the ELISA. The titer for the detection antibody was determined. The direct ELISA could determine the concentration of hirudin in the range of ~10ng/ml. Affinity pulified IgG was obtained and conjugated to horseradish peroxidase. Purified IgG and IgG-HRP could be used as capture and detection antibody, respectively. Although sandwich ELISA would not give the satisfactory results. it could apply for the detection of hirudin level in the range of ~20 ${\mu}$g/ml.