• Current Research on Agriculture and Life Sciences
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• v.1
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• pp.195-199
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• 1983

A microplate enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to bovine leukemia virus(BLV) is described and compared its sensitivity with that of the agar gel immunodiffusion test (ID) with BLV glycoprotein (gp) antigen using 263 sera collected in Korea and Japan. There was 98.5 per cent agreement between ELISA and ID when ELISA value, the value of tested serum(T) was devided with that of standard negative seurm(N) after the value of control was eliminated from T and N (T-C/N-C), of 1.5 or greater was considered positive. One hundred and forty four (99.6%) of 145 sera which were positive by ID were greater than 1.5 by ELISA, and 115 (97.5%) of 118 sera which were negative by ID were less than 1.5 by ELISA. As a result, it suggest that the ELISA test using BLV-gp antigen provides a useful serological tool for the diagnosis of BLV infection.

• Korean Journal of Veterinary Research
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• v.34 no.1
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• pp.93-97
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• 1994

A immunological survey of paratuberculosis in dairy and Korean native cattles was conducted by enzyme linked immunosorbent assay(ELISA), complement fixation test(CFT), agar gel immunodiffusion test (AGID) and intradermal skin test(ID). Over all prevalence of pararuberculosis in cattles was 6.7%(109/1633) by ID, 7.5(205/2719) by AGID, 9.3% (245/2641) by CFT and 13.4%(363/2719) by ELISA. Prevalence in dairy cattle was higher than that of Korean native cattle. Of 70 ELISA-positive cattle, 23(28.6%) and 48(68.6%) cattles were classified as positive in the AGID and positive or suspect in CFT, respectively. Of 92 ELISA-suspect cattle, 32(34.9%) and 48(52.2%) cattles were classified as AGID-positive and CFT-positive or suspect, respectively. It was concluded that paratuberculosis is widespread in cattle of Korea.

• Journal of Food Hygiene and Safety
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• v.31 no.1
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• pp.21-27
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• 2016

The objectives of this study are to produce monoclonal antibodies (MAbs) against Vibrio parahaemolyticus and to develop an immuno-selective filtration (ISF) method for the rapid and sensitive detection of V. parahaemolyticus. The characterization of the MAb produced from HKVP 4H9-9 hybridoma cell was validated by enzyme-linked immunosorbent assay (ELISA) and western blot. The produced MAb was specific to V. parahaemolyticus and showed weak cross-reaction to V. alginolyticus, V. vulnificus and Staphylococcus aureus. After optimization of the method, $5{\times}10^1cell/mL$ of V. parahaemolyticus in a pure culture could be detectable. Although weak cross-reactivity to V. vulnificus, V. alginolyticus and Staphylococcus aureus was observed, the ISF was confirmed to be highly specific to V. parahaemolyticus. Especially, the ISF showed the most sensitivity compared to the immunoassays currently reported is easier to perform and quicker than ID-ELISA.

• Journal of Food Hygiene and Safety
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• v.29 no.2
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• pp.158-163
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• 2014

In this study, two commercial PCR and ELISA test kits were examined for identification of eight animal species (beef, pork, chicken, duck, turkey, goat, lamb, and horse) from raw meat and meat products in Korea. The detection limit in RAW meat ELISA kit$^{(R)}$ on three types of meat samples blended with beef, pork and chicken, demonstrated that all meat species were differentiable down to 0.2%. RAW meat ELISA kit$^{(R)}$ on animal species resulted in differentiation rate of 94.5% for beef, 93.3% for pork, 90% for lamb, and 100% for chicken, duck, turkey, goat, and horse. In contrast, Powercheck Animal Species ID PCR kit$^{TM}$ resulted in 100% specificity at 0.05% limit of detection for all meat species. The detection limit of Cooked Meat ELISA kit$^{(R)}$ on mixed meat samples heat-treated with different temperatures and times, resulted in 0.1% for all heat-treated mixed meat except for chicken at 1.0%. Additionally, ELISA kit on sixty meat products resulted in specificity of 31.8% for ham, 13.6% for sausages, and 12.5% for ground processed products, and relatively low rate for more than 2 types of mixed meats. On the contrary, meat species differentiation using PCR kit showed higher percentage than that using ELISA kit$^{(R)}$: 50.0% for ham, 41.7% for sausages, and 28.6% for ground processed meat. Futhermore, PCR kit on 54 dried beef meats detected pork genes in 13 products whereas ELISA kit showed negative results for all products. Hence, the possibility of cross-contamination during manufacturing process was investigated, and it was found that identical tumblers, straining trays, cutters and dryers were used in both beef and pork jerky production line, suggesting the inclusion of pork genes in beef products due to cross-contamination. In this study, PCR and ELISA test kits were found to be excellent methods for meat species differentiation in raw meat and heat-processed mixed meat. However, lower differentiation rate demonstrated in case of meat processed products raised the possibility of inclusion of other species due to cross-contamination during manufacturing process.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.329-334
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• 2010

Escherichia coli O157:H7 causes hemolytic uremic syndrome and hemorrhagic colitis in humans. The objectives of this study were to produce monoclonal antibody(MAb) against E. coli O157:H7 and to develop an enzyme linked immunosorbent assay(ELISA) for the rapid detection of E. coli O157:H7 in agri-stockbreeding. The characterization of MAb produced from hybridoma cell (HKEC 4G8-5) was validated by ELISA and Western blot. The produced MAb was specific to E. coli O157:H7 and showed weak cross-reaction to Staphylococcus aureus. The detection limit of ELISA based on 4G8-5 MAb was $10^5\;cell/mL$. Although the ELISA could not detect E. coli O157:H7 in the meat and sprout samples inoculated with $1{\times}10^1\;cell$/10 g without enrichment, the same samples after enrichment for 6 hr were confirmed to be positive by ELISA. These results indicated that the ELISA combined with short enrichment (6 hr) is useful tool for rapid screening of E. coli O157:H7 in various samples.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1776-1781
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• 2012

Multiple viruses such as Lily symptomless virus (LSV), Lily mottle virus (LMoV), and cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf samples and bulbs showing characteristic symptoms of virus infection were collected from Gangwon, Chungnam, and Jeju provinces of Korea in 2008-2011. Coat protein (CP) genes of LSV and LMoV were amplified from collected samples by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into a pET21d(+) expression vector to generate recombinant CPs. The resulting carboxy-terminal His-tagged CPs were expressed in Escherichia coli strain BL21(DE3) by isopropyl-1-thio-${\beta}$-D-galactoside induction. The recombinant proteins were purified using Ni-NTA agarose beads, and the purified proteins were used as an immunogen to produce polyclonal antibodies in rabbits. The resulting polyclonal antisera recognized specifically LSV and LMoV from infected plant tissues in Western blotting assays. Indirect enzymelinked immunosorbent assay and immunocapture RT-PCR using these polyclonal antisera were developed for the sensitive, efficient, economic, and rapid detection of Lily viruses. These results suggest that large-scale bulb tests and economic detection of Lily viruses in epidemiological studies can be performed routinely using these polyclonal antisera.

• Research in Plant Disease
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• v.9 no.2
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• pp.94-98
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• 2003

To produce antibodies against Pseudomonas tolaasii and P. agarici, lyophilized P. tolaasii and P. agarici ($5{\times}10^7$ cfu/ml) and Freund, s adjuvant were immunized into rabbits 4 times. The specificity and sensitivity of the antibodies were evaluated by immunodiffusion test and indirect enzyme-linked immunosorbent assay (id ELISA). The ${\alpha}$-P. tolaasii antibody was very specific only against P. tolaasii, while ${\alpha}$-P. agarici antibody was not specific and showed a high cross reactivity toward P. tolaasii with detection limit concentration of $2{\times}10^3$ cfu/ml. However, the cross reactivities of ${\alpha}$-P. agarici antibody toward the related species including P. reactans were very low. Our results showed that ${\alpha}$-P. tolaasii and ${\alpha}$-P. agarici antibodies against P. tolaasii and P. agarici, respectively, might be useful for rapid and simple detection of the causal agents of bacterial brown and yellow blotches in cultivated oyster mushrooms.

• Journal of Veterinary Science
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• v.22 no.3
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• pp.29.1-29.6
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• 2021

West Nile virus (WNV), a neurotropic arbovirus, has been detected in mosquitos, birds, wildlife, horses, and humans in Malaysia, but limited information is available on WNV infection in Malaysian pigs. We tested 80 archived swine serum samples for the presence of WNV antibody and West Nile (WN) viral RNA using ID Screen West Nile Competition Multi-species enzyme-linked immunosorbent assay kits and WNV-specific primers in reverse transcription polymerase chain reaction assays, respectively. A WNV seroprevalence of 62.5% (50/80) at 95% confidence interval (51.6%-72.3%) was recorded, with a significantly higher seroprevalence among young pigs (weaner and grower) and pigs from south Malaysia. One sample was positive for Japanese encephalitis virus antibodies; WN viral RNA was not detected in any of the serum samples.

• International Journal of Oral Biology
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• v.42 no.1
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• pp.17-23
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• 2017

Background: Periodontitis is generally a chronic disorder characterized by the breakdown of tooth-supporting tissues. P. gingivalis, a Gram-negative anaerobic rod, is one of the major pathogens associated with periodontitis. Frequently, P. gingivalis infection leads to cell death. However, the correlation between P. gingivalis-induced cell death and periodontal inflammation remains to be elucidated. Among cell deaths, the death of immune cells appears to play a significant role in inflammatory response. Thus, the aim of this study was to examine P. gingivalis-induced cell death, focusing on autophagy and apoptosis in THP-1 cells. Methods: Human acute monocytic leukemia cell line (THP-1) was used for all experiments. Autophagy induced by P. gingivalis in THP-1 cells was examined by Cyto ID staining. Intracellular autophagic vacuoles were observed by fluorescence microscopy using staining Acridine orange (AO); and 3-methyladenine (3-MA) was used to inhibit autophagy. Total cell death was measured by LDH assay. Cytokine production was measured by an ELISA method. Results: P. gingivalis induced autophagy in an MOI-dependent manner in THP-1 cells, but 3-MA treatment decreased autophagy and increased the apoptotic blebs. P. gingivalis infection did not increase apoptosis compared to the control cells, whereas inhibition of autophagy by 3-MA significantly increased apoptosis in P. gingivalis-infected THP-1 cells. Inhibition of autophagy by 3-MA also increased total cell deaths and inflammatory cytokine production, including $IL-1{\beta}$ and $TNF-{\alpha}$. Conclusion: P. gingivalis induced autophagy in THP-1 cells, but the inhibition of autophagy by 3-MA stimulated apoptosis, leading to increased cell deaths and pro-inflammatory cytokines production. Hence, the modulation of cell deaths may provide a mechanism to fight against invading microorganisms in host cells and could be a promising way to control inflammation.

• Nuclear Medicine and Molecular Imaging
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• v.42 no.4
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• pp.307-313
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• 2008

Purpose: Vascular endothelial growth factor (VEGF) and its receptor, fetal liver kinase 1 (Flk-1), play an important role in vascular permeability and tumor angiogenesis. The aim of this study is to evaluate the therapeutic efficacy of $^{131}I$ labeled anti-Flk-1 monoclonal antibody (DC101) on the growth of melanoma tumor, which is known to be very aggressive in vivo. Materials and Methods: Balb/c nude mice were injected subcutaneously with melanoma cells in the right flank. Tumors were allowed to grow up to $200-250\;mm^3$ in volume. Gamma camera imaging and biodistribution studies were performed to identify an uptake of $^{131}I$-DC101 in various organs. Mice with tumor were randomly divided into five groups (10 mice per group) and injected intravenously; control PBS (group 1), $^{131}I$-DC101 $50\;{\mu}g/mouse$ (group 2), non-labeled DC101 $50\;{\mu}g/mouse$ (group 3), $^{131}I$-DC101 $30\;{\mu}g/mouse$ (group 4) and $15\;{\mu}g/mouse$ (group 5) every 3 or 4 days for 20 days. Tumor volume was measured with caliper twice a week. Results: In gamma camera images, the uptake of $^{131}I$-DC101 into tumor and thyroid was increased with time. Biodistribution results showed that the radioactivity of blood and other major organ was gradually decreased with time whereas tumor uptake was increased up to 48 hr and then decreased. After 4th injection of $^{131}I$-DC101, tumor volume of group 2 and 4 was significantly smaller than that group 1. After 5th injection, the tumor volume of group 5 also significantly reduced. Conclusion: These results indicated that delivery of $^{131}I$ to tumor using FlK-1 antibody, DC101, effectively blocks tumor growth in aggressive melanoma xenograft model.