• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.567-575
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• 1989

This study was conducted to evaluate the possibility of application of a microenzyme-linked immunosorbent assay(micro-ELISA) for the serodiagnosis of specific toxoplasma antibodies in swine sera and this test was performed as a microplate system by coating the polystyrene plates with toxoplasma soluble antigen, incubated serially diluted sera, then added horse radish peroxidase labelled goat anti-swine IgG(r) conjugate followed by o-phenylenediamine as substrate. The color development by enzyme-substrate reaction was determined by the photometric reading [ELISA reader at 490nm (OD)] and visual reading. The soluble antigen was prepared from the tachyzoites in mouse peritoneal cavity. A total of 1,200 swine sera from pig slaughter-house and a total of 116 swine sera from pig breeding station (S-C farm) were tested for the detection of antibodies to Toxoplasma gondii. The results obtained were summarized as follows: 1. The optimal reactions of indirect ELISA for the test sera were determined by the dilution of antigen 1:256 and 1:3,200 of horse radish peroxidase conjugate [anti-swine IgG(r)]. 2. The specific togoplasma antibody(IgG) in pigs infected with Tp artificially were detected as the serum titers of 1:64 or 1:128 at one week postinfection. 3. Of a total of 1,200 swine sera from pig slaughter-house 505 samples of sera were detected as positive (42.1%) and of a total of 116 swine sera from S-C pig breeding station 68 samples of sera as positive (58.6%). 4. The specific antibody(IgG) detection rates against a total of 1,200 test sera from pig slaughter-house were not significant between male (43.1%) and female (40.7%). 5. The indirect ELISA was proved to be a sensitive and specific procedure for the serodiagnosis of swine toxoplasmosis and also evaluated as an effective screening test for the large scale of test samples in laboratory.

• Biomedical Science Letters
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• v.2 no.1
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• pp.31-40
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• 1996

Escherichia coli is one of the most common etiological agents in urinary tract infection. An important virulence factor is the adhesive capacity of E. coli to uroepithelial cell, mediated by bacterial fimbriae. The Adhesion property has been regarded as an important virulence determinant in urinary tract infections. A total of 60 patients, who were diagnosed microbiologically as urinary tract infections, were examined by immunoblotting and enzyme-linked immunosorbent assay(ELISA). Uropathogenic E. coli with recombinant plasmid were positive for mannose resistant hemagglutination (MRHA). For identification of p-fimbriae subtype in uropathogenic E. coli, In the immunoblot analysis, specific bands in the range of p-fimbriae molecular weight of 17KD-22KD were identified. For the distribution of p-fimbriae subtype in the patient sera, 34/60(56.7%) were positive for $F7_1$, 28/60(46.7%) were positive for $F7_2$, and 30/60(50%) were positive for F13 with immunoblotting method. similar trends were observed in the enzyme-linked immunosorbent assay. Relatively good specificity(92.6%) and sensitivity(90%) were found in the ELISA test system using mixed antigens of purified $F7_1$, $F7_2$, and F13 p-fimbriae, and 60 sera from patients with urinary tract infections. In conclusion The serological tests were convenient method in diagnosis of urinary tract infections. among those ELISA could be recommended in diagnosis of urinary tract infections.

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

• Environmental Analysis Health and Toxicology
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• v.11 no.1_2
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• pp.11-17
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• 1996

The purpose of this experiment is to estabilish the sensitive and specific ELISA (enzyme linked immunosorbent assay) system for the detection of fish metallothionein (MT). Silver carp were injected with CA of 1-8mg/kg body wt. 4 times during 10 days. Silver carp was very tolerant species to CA. Cd induced MT in liver was seperated and purified by gel filtration chromatography and ion exchange chromatography and identified by spectrophotometry, native gel electrophoresis and western blot analysis. The rabbit antiserum was produced by immunizing rabbit with lyophilized MT, and the competitive ELISA system was estabilished for the detection of fish MT. In the present ELISA system, the detection limit was about 33 ng/ml. When this ELISA system was employed to determine the MT level in the supernatant sample of fish liver homogenate, the reaction curve showed a good parallel corelationship with the calibration curve over a certain dilution range. The results indicate that the competitive ELISA can be a useful tool for the detection of fish MT in the toxicological study and the evaluation of water pollution.

• Microbiology and Biotechnology Letters
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• v.27 no.4
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• pp.344-348
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• 1999

A competitive enzyme-linked immunosorbent assay(ELISA) for the detection of antibodies to Treponema pallidum(T.pallidum) was developed and evaluated. T.apllidum lysate was immobilized on the surface of microplate wells and horseradish peroxidase labeled human anti-T.pallidum lysate was immobilized on the surface of microplate wells and horseradish peroxidase labeled human anti-T.pallidum was prepared and used as a tracer. The performance of the competitive ELISA was evaluated by using different specimens. The competitive ELISA showed a sensitivity of 100% in a performance panel consisting of serum and plasma with anti-T.pallidum reactivity ranging from negative to strong positive by FTA-ABS test system and 120 plasma samples positive by TPHA. The specificity of the competitive ELISA was 100% in 1,200 plasma samples collected from healthy seronegative blood donors. These results suggest that the competitive ELISA provides an excellent assay method for the detection of antibodies to T.pallidum, and may be particularly useful for serological blood screening of syphilis.

• Korean Journal of Food Science and Technology
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• v.28 no.5
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• pp.953-958
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• 1996

A simple and rapid ELISA (enzyme-linked immunosorbent assay) system for fumonisins, a group of potentent carcinogen, was developed. To produce anti-fumonisin B1 (FB1) antibodies, FB1 conjugated to keyhole lympet hemocyanin (KLH) and Freund's adjuvant were immunized into rabbits subcutaneously 3 times. From one of the antisera showing high titer and good competition with the toxin in ELISA, polyclonal antibodies were purified. The cross-reactivities of the antibodies against fumonisin $B_1,\;B_2\;and\;B_3$ were 100%, 69%, and 166%, respectively. When competitive direct ELISA established by use of the antibody was applied to the spike test of $FB_1$ onto uncontaminated corns, the assay recovery was unstable unless 75% methanol extracts of corn were diluted to 1/100 with buffer. In that condition the mean ELISA recovery of FB1 from corns spiked $1-30\;{\mu}g/g$ was 67% and stable (coefficient of variation (CV) of each recovery percentage, 3.4%). The results suggest that the ELISA system established in this study needs no cleanup procedure and therefore would be powerful to screen a large number of corn samples contaminated with fumonisins.

• Korean Journal of Food Science and Technology
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• v.28 no.6
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• pp.1184-1187
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• 1996

A simple and rapid detection system for $aflatoxin\;M_1\;(AFM_1)$ in cow's milk by an enzyme-linked immunosorbent assay (ELISA) was developed. Specific antibodies against $AFM_1$, conjugated to bovine serum albumin $(AFM_1-BSA)$ were raised in rabbits and purified. The cross-reactivities of the antibodies against aflatoxin analogs were less than 29.9%. When a competitive direct ELISA (cdELISA) for $AFM_1$, established by use of the antibodies was applied to the spike test of $AFM_1$ onto uncontaminated cow's milk, the assay recovery was unstable unless cow's milk was diluted to 40% (2:3) with phosphate buffered saline (PBS). In that condition of sample dilution, the mean ELISA recovery of $AFM_1$, from the cow's milk was 113% (coefficient of variation (CV) of each recovery percentage, 8.2%) in the range of $0.3{\sim}3.0\;ppb$. These results showed that the ELISA system could be a convenient tool to monitor the contamination of AFM1 more than 0.5 ppb in cow's milk (FDA allowance limit) easily.

• Korean Journal of Food Science and Technology
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• v.32 no.2
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• pp.439-443
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• 2000

In order to evaluate the safety of greenhouse horticulture products in Korea, we carried out this work by screening of Fusarium species, which produce deoxynivalenol (DON) from greenhouse horticulture in Western Gyeongnam and Northern Gyeongbuk, Korea. For this study, high sensitive enzyme-linked immunosorbent assay, ALP/NADP method, was applied to detection of DON by enzyme amplification system. From 192 samples of greenhouse horticulture soil and its products, 103 isolates of Fusarium species were obtained. The isolates were cultured at 28C for 15 days and the cultured mediums were extracted by ethyl acetate. The production of DON was verified by thin layer chromatography (TLC). As the results of TLC, 8 strains were identified as DON producing strain. We screened potential producers of DON by ALP/NADP. The levels of DON production were shown from 0.007 to 1.21 g/ml of YES medium. The maximum DON producing strain No. 32-D-3 was isolated from soil in Namhae, Korea. In conclusion, the above results indicate that DON producing fungi contaminated greenhouse horticulture products in Korea. Therefore, further studies are required to accumulate more detailed data about the contamination of DON in various cereals.

• Bulletin of the Korean Chemical Society
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• v.26 no.6
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• pp.939-942
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• 2005

For the detection of cyanobacterial toxin, an Enzyme-linked immunosorbent assay (ELISA) was integrated into a PDMS microchip. The conjugates of microcystin-LR (MCLR) and keyhole limpet hemocyanin (KLH) were adsorbed on the surface of polystyrene beads and these MCLR-KLH polystyrene beads were introduced into a microchamber. MCLR on the surface of polystyrene beads reacted with horseradish peroxides (HRP) conjugated anti-MCLR monoclonal antibody (mAb) which had a competitive reaction with MCLR in water sample. After the enzyme substrate 3,3,5,5-tetramethyl benzidine (TMB) was injected into the chamber and catalyzed by HRP, the color change was detected with a liquid-cord waveguide. This integration shortened the conventional ELISA analysis time from several hours to about 30 min with only 4.2 $\mu$L MCLR sample consuming which was useful for the environmental analysis. More over, troublesome operations required for ELISA could be replaced by simple operations. The microchip based detection system showed a good sensitivity of 0.05 $\mu$g/L and maintained good reliability through its quantitative range with low coefficients of variation (2.5-10.5%).

• Journal of Periodontal and Implant Science
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• v.46 no.5
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• pp.320-328
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• 2016

Purpose: Human saliva, as a vital part of the immune defense system, contains a number of distinct proteins and peptides. Recently human common salivary protein 1 (CSP1) has been identified as an abundant salivary protein and may play a role in promoting the binding of cariogenic bacteria to salivary pellicles. However, nothing else is known regarding the role of CSP1 in periodontology. The aim of this study was to quantify and compare CSP1 levels between healthy subjects and periodontal patients. Methods: This controlled clinical study was conducted in periodontally healthy individuals and patients with chronic periodontitis Chonbuk National University Hospital, with Institutional Review Board approval. Whole saliva samples were collected from 36 healthy subjects and 33 chronic periodontitis patients and analyzed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immune blotting were conducted to ensure that anti-CSP1 monoclonal antibody (mAb) binds to CSP1 in human saliva. A sandwich enzyme-linked immunosorbent assay (ELISA) system was house-fabricated using mAb-hCSP1#14 and mAb-hCSP1#4 as a capture and a detector mAb, respectively. The CSP1 concentrations in saliva from 36 healthy subjects and 33 periodontal patients were quantified using the CSP1 sandwich ELISA system, and the results were analyzed using the Student's t-test. Results: Immunoblot analysis using mAb-hCSP1 as a probe confirmed that CSP1 in human saliva existed as a single band with a molecular weight of approximately 27-kDa. The quantification of CSP1 concentrations by CSP1 ELISA showed that the median values (25th to 75th percentiles) of periodontal patients and healthy subjects were 9,474 ng/mL (range, 8,434.10,139 ng/mL) and 8,598 ng/mL (range, 7,421.9,877 ng/mL), respectively. The Student's t-test indicated the presence of a statistically significant difference between the 2 groups (P=0.024). Conclusions: The presence of a significant difference in CSP1 levels between healthy subjects and periodontal patients suggests that CSP1 may be a potential biomarker for the detection or screening of periodontitis patients.