• Parasites, Hosts and Diseases
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• v.50 no.1
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• pp.15-21
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• 2012

In Iran, $Plasmodium$ $vivax$ is responsible for more than 80% of the infected cases of malaria per year. Control interventions for vivax malaria in humans rely mainly on developed diagnostic methods. Recombinant $P.$ $vivax$ apical membrane antigen-1 (rPvAMA-1) has been reported to achieve designing rapid, sensitive, and specific molecular diagnosis. This study aimed to perform isolation and expression of a rPvAMA-1, derived from Iranian patients residing in an endemic area. Then, the diagnostic efficiency of the characterized Iranian PvAMA-1 was assessed using an indirect ELISA method. For this purpose, a partial region of AMA-1 gene was amplified, cloned, and expressed in pET32a plasmid. The recombinant $His-tagged$ protein was purified and used to coat the ELISA plate. Antibody detection was assessed by indirect ELISA using rPvAMA-1. The validity of the ELISA method for detection of anti-$P.$ $vivax$ antibodies in the field was compared to light microscopy on 84 confirmed $P.$ $vivax$ patients and compared to 84 non-$P.$ $vivax$ infected individuals. The ELISA cut-off value was calculated as the mean+2SD of OD values of the people living in malaria endemic areas from a south part of Iran. We found a cut-off point of OD=0.311 that showed the best correlation between the sera confirmed with $P.$ $vivax$ infection and healthy control sera. A sensitivity of 81.0% and specificity of 84.5% were found at this cut off titer. A good degree of statistical agreement was found between ELISA using rPvAMA-1 and light microscopy (0.827) by Kappa analysis.

• Journal of fish pathology
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• v.12 no.1
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• pp.24-31
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• 1999

Optimization and standardization of solid phase enzyme immunoassay were done for the diagnosis of edwardsiellosis in fish. The analyzed degree of immobilized antibody on surface of solid phase with peroxidase saturation method showed the optimized result by using partially purified $50{\mu}g/ml$ of rabbit anti-E. tarda Edk-2 antibody in sodium bicarbonate buffer for overnight incubation to cover the surface of polystyrene beads. Optimized immunoreaction was observed in the treatment of $50{\mu}g/ml$ of biotin conjugated antibody followed extravidin-peroxidase diluted 1 : 2,000 in PBS. The detectable concentrations of the this method were $1{\times}10^5$ cells/ml and $1{\times}10^5$ cells/ml expressed as the source of antigen amount for EDTA extraction and heat extraction, respectively. High cross-reaction of solid phase ELISA with the prepared rabbit and-E. tarda Edk-2 was observed against E. tarda strains isolated from flounder suffering from edwardsiellosis in aquatic farms of Korea. It suggested that the potential of this solid phase of ELISA technique is very powerful for the application to different strains of E. tarda isolated in farms of many different areas.

• 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 Environmental Health Sciences
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• v.36 no.1
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• pp.52-60
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• 2010

High performance liquid chromatography (HPLC) and liquid chromatography mass spectrometry (LC/MS) have been widely used to quantify aflatoxins in food, but these methods are expensive, time-consuming, unsuitable for analysis of the routine screening of large sample numbers and require derivatization and high level techniques to perform. The objective of this study is to detect aflatoxins in a large number of foods by a high efficient analytical system of combined enzyme linked immunosorbent assay (ELISA) for screening and LC/MS/MS for confirmation. The samples spiked individually with aflatoxin $B_1$ (0.5 and 1.0 ng/g) and total aflatoxins (10 ng/g) were analyzed by ELISA and LC/MS/ MS, and the recoveries for ELISA and LC/MS/MS were 71.8~119.2% and 70.8~135.3%, respectively. A total of 378 samples (grains, nuts, soybean and fermented soybean foods, pepper and fermented pepper foods) were purchased from the six major cities in Korea and analyzed by ELISA-LC/MS/MS system. Twenty two (5.8%; peanut: 11, pistachio: 2, walnut: 6, almond: 1, pepper powder: 1, pepper paste: 1) out of 378 samples were screened as aflatoxin B1 positive by ELISA, but, 4 (1.1%; peanut: 2, pistachio:1, pepper powder: 1) out of the 22 samples screened were confirmed as aflatoxins positive at levels of 1.02~52.79 ng/g by LC/MS/MS. ELISA-LC/MS/MS system provides a more rapid, accurate and cost-effective method for the detection of aflatoxins in large number of samples.

• KSBB Journal
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• v.21 no.3
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• pp.212-219
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• 2006

The purpose of this study was to develop a assay system of host cell-derived residual proteins in final pharmaceutical products. Accurate and simple assay system for host cell-derived proteins(HCPs) is very important test item in pharmaceutical qualification control. In this study, methods for quantification of residual HCPs in recombinant anti-GPIIbIIIa antibody were developed using a process-specific immunoligand assay which was based on the Enzyme linked Immunosorbent assay(ELISA) system. The assay had a detection limit of 10.8 ng/ml of HCPs with a product concentration of 1 mg/ml. The practical implication of these results is that the developed ELISA system can be used for HCPs qualification control and this system will be applicable to develop another ELISA system of different antibody drug.

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

• Proceedings of the Korean Environmental Sciences Society Conference
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• 2003.11a
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• pp.99-102
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• 2003

The different methods such as HPLC, indirect- and direct-ELISA were employed for the analysis of microtoxins and the results of each method were compared in terms of the detection limit and accurary. Three toxins, microcystin-RR, -LR and -YR were clearly separated by HPLC using 0.05 M methanol and phosphate buffer used as a solvent system. The calibration curves for the toxins were linear in the range of 5 ng to 50 ng. The standard curves for the immunoassay of microcystin obtained by direct and indirect ELISA are compared. The linear responses of inhibitions of binding by microcystin in the direct and indirect competitive ELISA were in the range of 10 ng to 1000 ng and 50 pg to 160 pg, respectively. Distribution of microtoxins at 11 sites in the Nakdong river and several lakes in Korea was also studied. The most dominant microcystin variant in the test sites was found to be microcystin-RR.

• Bulletin of the Korean Chemical Society
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• v.24 no.3
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• pp.334-338
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• 2003

A simple synthetic method for haptens of organophosphorus (OP) pesticides with a spacer arm (aminocarboxylic acid) attached at the pesticide thiophosphate group was developed and was applied to the synthesis of haptens for the OP fungicide tolclofos-methyl. Using the haptens, a selective enzyme-linked immunosorbent assay (ELISA) for tolclofos-methyl was developed. One of the haptens was coupled to BSA to use as an immunogen. Rabbits were immunized with this conjugate to obtain polyclonal antibodies to tolclofos-methyl. The antisera were screened against another hapten coupled to ovalbumin (OVA). Using the serum with highest specificity, an antigen-coated ELISA was developed, which showed an $IC_{50}$ of 160 ng/mL with the detection limit of 20 ng/mL. The antibodies showed negligible cross-reactivity with other OP pesticides. An antibody-coated ELISA was also developed, which showed an $IC_{50}$ of 410 ng/mL with a detection limit of 130 ng/mL.

• Korean Journal of Veterinary Research
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• v.30 no.2
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• pp.187-192
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• 1990

The Presence and quantity of protein A in Staphylococcus hyicus subsp hyicus isolates from pigs were determined by indirect hemagglutination test and enzyme-linked immunosorbent assay(ELISA). Cell-bound and extracellular protein A was demonstrated in 87.7% and 36.0% of 489 isolates, respectively, by indirect hemagglutination test. When contents of the protein A were estimated by ELISA method, all of the isolates that were positive to protein A in indirect hemagglutination test produced extracellular protein A of less than 1ng/ml. Most of the isolates produced cell-bound protein A of less than 1ng/ml, whereas 28 isolates produced 1 to 35ng/ml and 11 isolates produced 25 to 108ng/ml.

• Research in Plant Disease
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• v.25 no.3
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• pp.131-135
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• 2019

Cylindrocarpon destructans causes ginseng root rot and produces radicicol that has an antifungal effect. In this study, we developed a method to detect this fungus using enzyme-linked immunosorbent assay (ELISA). Secreted proteins of C. destructans were used as antigens to obtain C. destructans-specific IgG from mouse. Out of 318 monoclonal antibodies generated from mouse, two antibodies (Cd7-2-2 and Cd7-2-10) showed highest specificity and sensitivity. Indirect ELISA using both antigens successfully detected C. destructans in soils, but direct ELISA using IgG conjugated with horseradish peroxidase failed to detect antigens in soils. The indirect ELISA developed here can efficiently detect the fungus and help manage ginseng root rot disease in fields.