• Reproductive and Developmental Biology
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• v.34 no.3
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• pp.161-167
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• 2010

Early diagnoses of pregnancy for animal such as swine and bovine is extremely important to increase income of a farmhouse and for the management of farm. For the development of immunoasaay system of pregnancy in swine, we report a competitive heterologous enzyme linked immunosorbent assay (ELISA) for the direct measurement of oestrone sulfate (E1S) in diluted urine using anti-E1G (glucuronide) monoclonal antibody which cross react with ElS. The principle of assay was based on the typical solid-phase competitive ELISA methods using E1G-HRP (horseradish peroxidase) as a tracer and E1S for standard. The method had a reasonable sensitivity for the detection of E1S with 0.15 ng/ml as a detection limit. The intra-assay and inter-assay precisions were raging coefficient of from 8.50~9.67% and 8.50~9.87%, respectively, which were quite acceptable. In a field trial with a group 37 sows (18 non-pregnancy and 19 pregnancy sows) after day 29~30 post service, the concentration of E1S were determined to be below 30 ng/ml in all non-pregnancy group and over 48 ng/ml in pregnancy group except one sample. The method described here, heterologous ELISA for the measurement of E1S in urine is good enough for monitoring the early pregnancy test of swine.

• Korean Journal of Environmental Agriculture
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• v.17 no.1
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• pp.22-25
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• 1998

These studies were conducted to develope analysis method of herbicide quizalofop-ethyl by Gas Liquid Chromatography(GLC) and Enzyme-Linked Immunosoment Assay(ELISA) in soil and plant. Quizalofop produced by hydrolysis of quizalofop-ethyl was conjugated with bovine serum albumin(BSA). Quizalofop antibody was developed in rabbits by using BSA conjugation. Antibody titer, incubation temperature, and incubation time was 32,000, $37^{\circ}C$ and 4hours respectively. Minimum detection limit of quizalofop-ethyl by ELISA was 5ppb. Quizalofop-ethyl recovery from soil by ELISA was more than 95percent. Minimum detection limit of quizalofop-ethyl by GLC was 5ppb. Quizalofop-ethyl recovery from soil by GLC was from 89 percent to 100 percent. Minimun detection limit of quizalofop-ethyl by HPLC was 100ppb. Quizalofop-ethyl recovery from soil by HPLC was 89.6 percent.

• Korean Journal of Food Science and Technology
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• v.46 no.5
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• pp.538-543
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• 2014

A sandwich enzyme-linked immunosorbent assay method (sELISA) for detecting the presence of shrimp in processed foods was developed using rabbit polyclonal antibodies against tropomyosin produced by black tiger prawns (shrimp). Based on the standard curve derived using this method, the detection range of shrimp was determined to be $1-100{\mu}g/mL$. The cross-reactivity of these antibodies toward black tiger prawns, fleshy prawns, cocktail prawns, lobster, and blue crab was 100, 73, 155, 18, and 0%, respectively. When shrimp was heated for 10 min, the mean assay recovery of tropomyosin was 121-221% at $70-100^{\circ}C$ and 7.8% at $121^{\circ}C$. When shrimp was added to cream soup, weaning food, sausage, fish paste, and sauce, the mean assay recovery was 397, 639, 168, 234, and 0%, respectively. In sample tests involving 14 commercial items, the coincidence ratio of assay results and reference was 79%.

• Journal of Food Hygiene and Safety
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• v.12 no.4
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• pp.281-287
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• 1997

We have reported a sensitive, specific and simple direct competitive ELISA method to detect aflatoxin in agricultural commodities. We evaluated the ELISA for practical use to detect aflatoxins contaminated in the domestic and foreign agricultural commodities. The detection limits of the direct ELISA for residual aflatoxins in rice, pine nuts, corns, almonds, bean nuts, and pistachio were 10 ppb and in peanuts and cashew nuts were 20 ppb, which were elucidated from the standard curves of ELISA for aflatoxin fortified into the agricultural commodities. Residue studies of naturally contaminated aflatoxins in the agricultural commodities were also carried out by using direct ELISA. As the results of the studies, it was revealed that there were no residues of aflatoxins in 20 rice samples produced in south Korea, 20 pine nut samples in south Korea (9 samples), USA (1 sample) and China (10 samples), each of 20 almond, pistachio and bean nut samples in USA. However, aflatoxin residues were detected in corn samples imported from north Korea (350∼585 ppb in 2 of 3 samples), from USA (109*326 ppb in 6 of 6 samples) and domestic corns (61-326 ppb in 7 of 17 samples). The toxins were contaminated in corn imported from USA for popcorn (17∼20 ppb, in 3 of 10 samples) whereas no residues were detected in corn from south Korea and China. In case of cashew nuts imported from India, 11.4∼23.1 ppb of aflatoxins were detected in 4 from 20 samples. Most of the contaminated foods were harvested before 1995. Thus, hygienic managements of the foods should be required during storage and circulation at market.

• Toxicological Research
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• v.19 no.2
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• pp.133-139
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• 2003

Allergic contact dermatitis may be caused by a wide variety of chemicals. A murine local lymph node assay (LLNA) has been developed as an alternative to guinea pig models for assessing the contact sensitization potential of chemical. However, there is a need to develop a nonradioisotopic endpoint for the LLNA, because of the radioisotopic method's requiring the use of special facilities. In this study, we investigated the development of a nonradioisotopic endpoint for LLNA using ELISA (enzyme-linked immunosorbent assay). Female Balb/c mice were treated by the topical application on the dorsum of both ears with four different strong sensitizers, 2,4-dinitrochlorobenzene (DNCB), oxazolone (OXZ), toluene diisocyanate (TDI), and trimellitic anhydride (TMA), and a strong irritant, sodium lauryl sulfate (SLS), once daily for three consecutive days. The proliferation of cells in the auricular Iymph node was analyzed by means of the labelling index (Ll) of bromodeoxyuridine (BrdU) incorporation into cells. The weights of the Iymph nodes in the mice treated with allergens, DNCB, OXZ, TDl and TMA were increased compared to the vehicle control. The stimulation index (Sl) of mice treated with DNCB, OXZ, TDl, and TMA was over three-fold increase compared to the vehicle control. However, the S1 of mice exposed to SLS was not significantly increased compared to the vehicle control, while the lymph node weight of SLS was significantly increased. These results suggest that the LLNA modified endpoint using ELISA based on BrdU incorporation could provide a useful method of screening for irritants and allergens.

• Journal of Veterinary Clinics
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• v.27 no.4
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• pp.402-406
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• 2010

Bovine leukemia virus (BLV) is a delta-retrovirus which causes chronic lymphocytosis in cattle. BLV infections have been divided into two groups such as enzootic bovine leukosis (EBL) and sporadic bovine leukosis (SBL) according to the clinical symptoms in infected cattle. The conventional detection method of BLV was hematological procedure which is determining lymphocytosis in the suspected animals. Recently several sensitive methods were developed to detect antibody to BLV and nucleic acid of the BLV from infected cattle. In this study we have compared the difference of positive rates between agar gel immunodiffusion (AGID) and enzyme linked immunosorbent assay (ELISA) which are using for BLV antibody detection methods. The positive detection rate of ELISA test was 7.4% greater than the positive rate of AGID. The discrepancy of the positive rate between ELISA and AGID were showed in the group of age over one year old to under three year old group. The result from each test agreed very well in the group of over 5 year old cattles. The serological test is very useful method to select the infected cattle for the eradication or control of the disease in the infected herd. But it has a limit by interference of the maternal antibody from the cow of under 6 month old. This study shows that 16.2% of these ages group showed BLV gene positive by polymerase chain reaction (PCR) method. The result suggests that ELISA test need to be used with PCR to clarify misinterpretation of positive animals by antibody response due to the natural infection from maternally derived antibody in calves of under 6 months old.

• Toxicological Research
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• v.17
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• pp.197-199
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• 2001

Enhancement of antigen-specific IgE is a hallmark of allergic hyperresponsiveness, therefore it is necessary to adopt or develop a highly sensitive and specific assay for determination of allergen-specific IgE levels in vivo. In this presentation, we introduce an ELISA (enzyme linked immunosorbent assay) system developed to measure the levels of chicken egg ovalbumin (OVA)-specific IgE in serum. The ELISA method uses a commercially available purified rat anti-mouse IgE as a capture Ab and biotinylated OVA as a detection reagent. Avidin-peroxidase with its substrate is used for color development resulting in optical density measurement at 405 nm. The ELISA system produces a highly sensitive dose-response relation-ship between optical density levels and the dilution titer of the OVA-IgE standard serum but no cross-reaction with unrelated IgE or IgG. It is believed that the system is an Efficient tool to delineate an adjuvant effect of environmental pollutants on development of asthmatic and atopic responses.

• Fisheries and Aquatic Sciences
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• v.4 no.1
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• pp.32-38
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• 2001

Several molecular biological techniques have been used to detect virus rapidly and accurately, but these methods have limitations in the early stage of viral infection with very low concentration of virus. We compared the detection limits of IMS-PCR, Western blot and ELISA with infectious hematopoietic necrosis virus OHNV). Four antibodies, rabbit anti-IHNV polyclonal antibody, anti-IHNV nucleocapsid protein monoclonal antibody, anti-IHNV nucleocapsid protein polyclonal antibody, and anti-IHNV glycoprotein polyclonal antibody, were tested to find out the most effective antibody for each method. The detection limit with IMS- PCR was $2\times10^6$ pfu when the viral RNA was extracted before RT-PCR. In the western blot with rabbit anti­IHNV polyclonal antibody one pfu of virus could be detected. In ELISA, 10 pfu of virus particles were detected with the same antibody.

• Archives of Pharmacal Research
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• v.10 no.1
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• pp.18-24
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• 1987

Mouse monocolonal antibodies to Hepatitis B surface antien (HBsAg) were prepared and their functional capabilities tested by the method of solid phase enzyme linked immuno sorbent assay (ELISA). HBsAg binding studies inicated that one monoclonal antibody 6E-1-1 bound more HBsAg at a faster rate than the other monoclonal antibodies. Also, for the binding inhibition studies with the selected monoclonal antibody 6E-1-1, one monoclonal antibody 8D-3-6 didn't exhibit binding inhibition for HBsAg. Then, a simultaneous ELISA method was developed for the immunodiagnosis of HBsAg. Different combinations of two monoclonal antibodies as solid phase and horseradish peroxidase (HRPO) labeled phase were studied. The combination of monoclonal antibody of higher affinity constant (6E-1-1) immobilized in a solid phase and monoclonal antibody of lower affinity constant (8D-3-6) as a HRPO laeled phase was more sensitive when two monoclonal antibodies of different affinity constants for HBsAg were prepared.

• Korean Journal of Veterinary Service
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• v.27 no.3
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• pp.273-280
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• 2004

The studies were performed for the PRRS antigen and antibody detection from breeding farms, artificial insemination(AI) center and growing farms in Jeonbuk province. 1. Specific PRRS primers were successfully amplified ORF6 617bp and ORF7 448 bp on agarose gel. 2. RT-PCR method has been establish by commercial kit and the thermal cycler program consisted of 30 cycles: $95^{\circ}C$ for 30 sec, $45^{\circ}C$ for 30 sec, and $72^{\circ}C$ for 45 sec. 3. The results of PRRS antibody test by ELISA method in AI centers were $6.6\%,\;53.3\%$ and breeding farms $65\%,\;65\%\;and\;38.7\%$, respectively. The serological positive of the antibody in gilt higher than sow. 4. The sero-positive of the PRRS antibody showed average $21\%$ in domestic farms, $56.2\%$ in breeding farms, and $29.9\%$ in AI center.