• Korean Journal of Veterinary Service
• /
• v.19 no.1
• /
• pp.30-38
• /
• 1996

Enzyme-linked immunosorbent assay (ELISA) was developed to detect rotavirus antigen in fecal samples using VP6-specific monoclonal antibody(2B12). The ELISA for rotavirus antigen detection found to have specificity to all bovine and porcine rotaviruses tested but not to bovine viral diarrhea virus and bovine coronavirus. The ELISA appeared to have similar sensitivity and specificity compared to fluorescence antibody assay(FA) and electropherotyping (PAGE).

• Parasites, Hosts and Diseases
• /
• v.47 no.2
• /
• pp.153-157
• /
• 2009

Diagnosis of hydatidosis is based on immunodiagnostic methods along with radiological and ultrasound examinations. The objectives of the present study were to develop a specific and simple antigen-based ELISA method for diagnosis of hydatidosis and compare it with antibody detection method. The subjects in this study included 89 patients in the following groups: surgically confirmed hydatidosis patients (35 cases), control with other parasitic diseases (29 cases), and healthy controls (25 cases). Hyperimmune serum was raised against hydatid cyst fluid in rabbits. Anti-hydatid cyst IgG was purified by affinity chromatography using protein A column and labeled with horseradish peroxidase. Collected sera were assessed for hydatid cyst antigens and antibody by ELISA. Circulating hydatid antigen was found in 9 out of 35 patients with surgically confirmed hydatidosis. A sensitivity of 25.7% and a specificity of 98.0% were calculated for the antigen detection assay. Antibody detection by indirect ELISA, using antigen B, showed that 94.2% of patients (33 cases) have anti-hydatid cyst antibodies in their serum while cross reaction was noted in a few of non-hydatidosis patients. A sensitivity of 94.2% and specificity of 81.6% were found for the antibody detection assay. Findings of this study indicated that antibody detection assay is a sensitive approach for diagnosis of hydatid cyst while antigen detection assay might be a useful approach for assessment of the efficacy of treatment especially after removal of the cyst.

• International Journal of Industrial Entomology and Biomaterials
• /
• v.6 no.2
• /
• pp.179-181
• /
• 2003

$F(ab`)_2$-ELISA and direct antigen coating-ELISA (DAC-ELISA) were evaluated in the detection of purified Bombyx mori nuclear polyhedrosis virus (BmNPV) and nuclear polyhedrosis virus infection in silkworm larvae inoculated with BmNPV polyhedra. Although nanogram levels of BmNPV was detected in both DAC- and $F(ab`)_2$-ELISA, similar concentrations of antigen was detected in case of F(ab’)$_2$-ELISA even at higher dilution of antibody (up to 1 : 20 K). One hundred percent nuclear polyhedrosis infection was detected 6 hrs after inoculation in BmNPV infected silkworm larvae by $F(ab`)_2$-ELISA. On the other hand, detection of 100% infection was observed only three days after inoculation in DAC-ELISA. In this study, it was observed $F(ab`)_2$-ELISA was more sensitive than DAC-ELISA in the detection of purified BmNPV as well as nuclear polyhedrosis infection in silkworm larvae.

• Korean Journal of Veterinary Research
• /
• v.42 no.1
• /
• pp.73-80
• /
• 2002

An enzyme-linked immunosorbent assay (ELISA) using purified hemagglutinin of swine influenza virus (H1N1) as antigen was developed for detection of antibody to avian influenza virus (AIV). The sensitivity and specificity of a developed and commercial available ELISA kits were compared with those of agar gel precipitation (AGP) test and hemagglutination inhibition (HI) test using sera collected from chickens under condition of field exposure. The concentration of antigen, serum dilution and concentration of enzyme-conjugated secondary antibody in developed ELISA (S-ELISA) were 0.5ug/100ul, 1:200 and 0.03ug/100ul, respectively. The correlation coefficients between S-ELISA and commercial ELISA and HI titers were 0.419 and 0.533, respectively. A significant correlation (p < 0.01) was not found between HI and ELISA titers. The S-ELISA was found to be as more sensitive and specific than the AGP test, showing 86.8% sensitivity and 85.3% specificity. It is suggested that the ELISA using the SIV as antigen may be useful method as an investigating tool for AIV serological surveillance.

• Korean Journal of Veterinary Service
• /
• v.21 no.3
• /
• pp.313-323
• /
• 1998

In order to establish a sensitive and specific diagnostic method for detection of antibody to Salmonella pullorum, a enzyme-linked immunosorbent assay(ELISA) was designed and standardized. The diagnostic efficacy of the established ELISA was compared with that of the serum plate agglutination test and immunodiffusion test for pullorum disease. 1. The chicken hyperimmune sera to Salmonella pullorum, S gallinarum, S typhimurium and S typhi were shown the cross reaction to S pullorum antigen by serum plate agglutination test. 2. When compared the cross reaction titer of microplate agglutination test for chickens hyperimmune sera, it was found that the titer were 64 in S pullorum, 32 in S gallinarum, 4 in S typhimurium and 8 in S typhi, respectively. 3. When compared the specificity of various antigen(HA, EA, PA and SA) by the immunodiffusion test, the most suitable antigen was phenol-treated bactrium. 4. The optimal concentration of S pullorum antigen for ELISA was 1 : 160 dilution of bacterium. 5. The efficacy of the ELISA for detection of S pullorum antibody was compared with serum Plate agglutination test and immunodiffusion test in chickens infected with S pullorum. The antibody was first detected at 6 days after infection using three tests examined. The antibody was alldetected at 9 days by ELISA, at 12 days by serumplate agglutination test, at 15 days by immunodiffusion test.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.18 no.3
• /
• pp.874-879
• /
• 2004

Alakaline phosphatase (ALP)- or horseradish peroxidase (HRP)-antibody conjugate was used frequently on the immunological detection methods such as enzyme-linked immunosobent assay (ELISA), immunobolt, immunohistochemistry. The classical enzyme-antibody coupling method by one-step (direction) injection of glutaraldehyde bring into being disadvantage such as low sensitivity of antigen detection because of homopolymers. This study was modified with the dialysis glutaraldehyde method to provide simple coupling through E-amino residues present in most protein. The dialysis glutaraldehyde coupling effects were better than the classical one-step glutaraldehyde injection in antigen detection of ELISA and immunobolt. Optimal dose of the dialysis glutaraldehyde solution was 0.10-0.25 %. This results suggest that the dialysis glutaraldehyde coupling method can readily applied to antigen detection of in vitro and in vivo.

• Korean Journal of Veterinary Service
• /
• v.34 no.2
• /
• pp.159-166
• /
• 2011

This study was carried out to develope enzyme-linked immunosorbent assay (ELISA) for detection of canine brucellosis in dogs experimentally inoculated with Brucella abortus 1119-3 and B. canis RM666. Groups A, B and C of dogs (each group consisting of three dogs) were orally inoculated with approximately $5{\times}10^9$ colony-forming units of B. abortus and B. canis, and with sterile pyrogen-free PBS, respectively. The animals were monitored at regular intervals upto the 12th week post inoculation (PI) by standard tube agglutination test (STAT), plate agglutination test (PAT), Rose Bengal test (RBT), 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and ELISA. The induced antibody titers in group A dogs were detected from the first week PI to the eighth week PI in STAT, PAT and RBT using the inactivated whole cells of B. abortus 1119-3 as antigens, while no sera in groups B and C dogs reacted with the antigens. In 2ME-RSAT using whole cells of B. canis M-strain as antigens, the induced antibody titers in group B dogs were observed at the second week PI and persisted for the 12th week PI, while sera of groups A and C dogs did not react with the whole cells. In ELISA using cytoplasmic fractions antigen of B. abortus 1119-3, the mean optical density of antibodies in groups A and B was detected from the first and second weeks PI, respectively, and persisted for 12th week PI, while sera of group C did not cross-react with the fractions antigen. However, in ELISA using the hot saline extracts of B. canis M- as an antigen, the induced antibody titers in only group B dogs were detected from second week PI and persisted for until the end of this study. These results indicate that the ELISA using B. abortus 1119-3 cytoplasmic fractions as antigens can be a good candidate for detection of brucellosis by B. abortus as well as B. canis in dogs.

• Biomedical Science Letters
• /
• v.21 no.1
• /
• pp.9-14
• /
• 2015

Mouse hepatitis virus (MHV) is a major pathogen in laboratory mice that usually leads to fatal diseases, such as hepatitis, multiple sclerosis, encephalitis, and respiratory disease. MHV has a high infection rate, and it needs to be detected as soon as possible to prevent its spread to other facilities. However, MHV detection by enzyme-linked immunosorbent assay (ELISA) often gives false positives; thus, it is very important that the results are confirmed as true positives in the early infection stage or distinguished as false positives with more accurate, reliable methods. Under microbiological screening, MHV ELISA-positive mice were found in four GFP-tagging transgenic mice. To verify the detection of the MHV antigen directly, reverse transcription polymerase chain reaction (RT-PCR) was performed, and the mice were determined to be MHV negative. Additional serum antibody-based screening was conducted with three different ELISA kits, and multiplexed fluorometric immunoassay (MFIA) was performed to confirm their accuracy/sensitivity. In brief, the ELISA kit for A59 nucleocapsid protein (MHV-A59N) revealed MHV ELISA positivity, while other ELISA kits (MHV-S lysate and MHV-JHM lysate) demonstrated MHV negativity. In MFIA, only the test for the recombinant A59 nucleocapsid antigen was MHV positive, which was consistent with the ELISA results. These results suggest that the ELISA kit with the recombinant A59 nucleocapsid antigen might induce non-specific MHV ELISA positivity and that confirmation is therefore essential.

• Korean Journal of Veterinary Research
• /
• v.45 no.4
• /
• pp.569-574
• /
• 2005

Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.

• Korean Journal of Veterinary Service
• /
• v.23 no.3
• /
• pp.271-279
• /
• 2000

This study was carried out to provide the fundamental information for development of proper vaccination program against infectious bursal disease(IBD) to the local chicken farms. The antigen detection was peformed from 8 samples of bursa of Fabricius with agar gel precipitation(AGP) and indirect immunofluorescent assay(IFA), And also, the antibodies in serum samples were detected by the various serological methods such as commercial ELISA assay, AGP and virus neutralization(VN) test. 1. The antigen detection rates were 25% for AGP which is 2 out of 8 farms and 10 out of 40 bursas, and 25% which Is 2 out of 8 farms and 20% 8 out of 40 bursas for IFA, respectively. 2. The mean titer of maternal antibody (>3,000) existed until 10 days of the age with ELISA-GMT. 3. The antibody positive rates which are over 80% showed until 5 days of the age with ELISA and at 10 days of the age with AGP except one, but none of them showed from 1 day of the age. This report came to conclusions that both the protective maternal antibody titers and the antigen positive rates were significant until at the 10 days of the age.