• Korean Chemical Engineering Research
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• v.49 no.4
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• pp.387-392
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• 2011

Enzymes are being used in various fields due to their unique property of substrate specificity. Enzyme-linked immunosorbent assay(ELISA) has enabled the detection of various antigens by reporting the binding event of antigen and antibody via enzyme-catalyzed reaction. However, the sensitivity improvement of conventional ELISA has been limited because only one enzyme molecule is conjugated to one molecule of antibody. To overcome this limitation and further improve the sensitivity of ELISA, there have been efforts to increase the number ratio of enzymes to antibody. Recently, the nanobiocatalytic approaches, with their successful enzyme stabilization, improved the performance stability as well as sensitivity in a modified protocol of ELISA. The present paper introduces the basic principle of ELISA, and the recent efforts to improve sensitivity and performance stability of ELISA by using the nanobiocatalytic approaches.

• Archives of Pharmacal Research
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• v.20 no.1
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• pp.46-52
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• 1997

To obtain more sensitive immunoassay for methamphetamine (MA) determination, the optimum condition of enzyme-linked immunosorbent assay (ELISA) was investigated in regard to immunogens, antibody purification methods and coating tracers. Activated MA, N-(4-aminobutyl)methamphetamine (4-ABMA), was conjugated with bovine serum albumin (BSA) or keyhole limpet hemocyanin (KLH) and used as immunogen. The antibodies were purified by protein G chromatography or various immunoaffinity chromatography-linked MA-protein ligands, such as MA-BSA, MA-KLH or MA-ovalbumin (OVA). Each purified antibody was characterized by means of sensitivity and cross-reactivity using the three MA-protein coating tracers, MA-BSA, MA-KLH and MA-OVA. The best sensitivity of each antibody was acquired with the MA-OVA tracer although the tracer concentration and the antibody titer level at optimum condition were varied. The antibody with high titer level did not always yield good sensitivity. At optimum condition, immunoaffinity chromatography-purified antibodies were better for sensitivity and for specificity than protein G-purified antibodies. The cross-reactivity of the purified antibodies seemed to be affected by immunogen structure and showed somewhat different patterns according to the immunoaffinity ligand utilized. These data show that the antibody purification method as well as choice of coating tracer and immunogen is essential for the sensitivity and specificity of EIA; the optimum condition for assay should be discovered using various methods and combinations.

• Parasites, Hosts and Diseases
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• v.28 no.1
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• pp.25-30
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• 1990

The direct wet mount examination of vaginal. secretion, widely applied for the diagnosis of Trichcmonas vaginalis infection in woman patients, is rapi4 and economical, however, the sensitivity of this technique is not so high. In this study enzyme-linked immunosorbent assay (ELISA) was employed for the detection of serum anti-T. vaginalis IgG and IsM antibodies from 30 vaginal trichomoniasis patients and 30 non-infected healthy persons. The results were as follows: 1. Serum ELISA-IgG value was $0.37{\pm}0.134(Mean{\pm}S.D.)$ in vaginal trichomoniasis patients and $0.21{\pm}0.054$ in healthy controls(P<0.005), and the sensitivity and specificity of ELISA for serum IgG antibody were 70.0% and 96.7%, respectively. 2. Serum ELISA-IgM value was $0.33{\pm}0.177 (Mean{\pm}S.D.)$ in vaginal trichomonlasls patients and $0.11{\pm}0.051$ in healthy controls (p<0.005), and the sensitivity and specificity of ELISA for serum IsM antibody were 70.0% and 96.7%, respectively. 3, The ELISA-IgG values showed a significant correlation with ELISA-IgM values(r=0.77, p<0.005) , With above results, it is assumed that ELISA is a reliable method for the diagnosis of T vaginalis infection and simultaneous measurement of serum IgG and IgM with this technique is recommended.

• Korean Journal of Veterinary Research
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• v.38 no.2
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• pp.314-318
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• 1998

An establishment of effective control measures to PRRSV infection in swine industry depends on a sensitive and specific diagnosis to detect either viral antigen and/or antibodies to PRRSV. Several diagnostic methods are available to detect antibodies against PRRSV, including IPMA, IFA and ELISA tests have been successfully developed. Sensitivity of the indirect immunofluorescent assay in MA-104 cells using Korean field isolate PL96-1 was superior to that of VR-2332 and field isolate PL96-2. Sensitivity and specificity of the IFA test with PL96-1 were comparable to those of commercial ELISA test kit but ELISA test was more sensitive for the detection of declining antibodies to PRRSV in finishing pigs. In this study we concluded that IFA and ELISA test could be utilized to detect antibodies to PRRSV and the results generated from these two tests were comparable and there were no significant difference between these two tests.

• Journal of Microbiology and Biotechnology
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• v.33 no.9
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• pp.1170-1178
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• 2023

Food allergy represents a severe problem for many societies, including sensitive populations, academies, health authorities, and the food industry. Peanut allergy occupies a special place in the food allergy spectrum. To prevent consumption by consumers suffering from a peanut allergy, a rapid and sensitive detection method is essential to identify unintended peanut adulteration in processed foods. In this study, we produced four monoclonal antibodies (MAbs; RO 3A1-12, PB 4C12-10, PB 5F9-23, and PB 6G4-30) specific to thermo-stable and soluble proteins (TSSPs) of peanut and developed an enzyme-linked immunosorbent assay (ELISA) based on the MAbs. Among them, PB 5F9-23 MAb was firmly bound to Ara h 1, and other MAbs strongly reacted to Ara h 3 in the Western blot analysis. An antibody cocktail solution of the MAbs was used to enhance the sensitivity of an indirect ELISA, and the limit of detection of the indirect ELISA based on the antibody cocktail solution was 1 ng/ml and improved compared to the indirect ELISA based on the single MAb (11 ng/ml). The cross-reaction analysis revealed the high specificity of developed MAbs to peanut TSSPs without cross-reaction to other food allergens, including nuts. Subsequently, analyzing processed foods by indirect ELISA, all foods labeled as containing peanuts in the product description were confirmed to be positive. The results indicate that the developed antibodies exhibit high specificity and sensitivity to peanuts and can be used as bio-receptors in immunoassays or biosensors to detect intentional or unintentional adulteration of peanuts in processed foods, particularly heat-processed foods.

• Korean Journal of Veterinary Service
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• v.21 no.2
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• pp.107-115
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• 1998

In order to establish a rapid, sensitive and specific diagnostic method for detection of antibody to Brucella abortus, a enzyme-linked immunosorbent assay(ELISA) was adapted. The diagnostic efficacy of the established ELISA was compared with that of the standard tube agglutination test for B abortus. 1. It was found that the optimal concentration of antigen for this ELISA was 5$\mu\textrm{g}$/ml, the optimal dilution of conjugate was 1 : 2000, and the optimal dilution of serum was 1 : 200, respectively. 2. Cut off value in this ELISA was 1,102 that was determined by mean absorbance(at 492nm) of tube agglutination test negative serum added with the triple value of the standared devation. 3. The relationship between the tube agglutination test and ELISA was showen high corresponding rate with sensitivity(96.3%) and specificity(98.1%). 4. The efficacy of the ELISA for detection of B abortus antibody was compared with tube agglutination test In brucellosis outbreak farm. The sensivity of ELSIA was higher than tube agglutination test.

• Korean Journal of Veterinary Research
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• v.42 no.1
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• pp.81-88
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• 2002

The purpose of this study was to conduct diagnosis of bovine paratuberculosis in Kangwon area. Blood samples were collected from 2,261 dairy cows of 162 herds, and the ELISA and immunoblotting using recombinant 34KDa protein of M. paratuberculosis were conducted. The feces collected from dairy cows were cultured on HEY medium with mycobactin-J and PCR was conducted with washing solution of medium 4 weeks after culture. The ELISA had sensitivity of 83.3% and specificity of 96.7%. And the immunoblotting had sensitivity of 83.3% and specificity of 100%. Of the 2,261 dairy cows, 371 cows(16.4%) were positive in ELISA and 75 cows(3.3%) were positive in immunoblotting. And of the 162 herds, 109 herds(67.3%) had an apparent paratuberculosis prevalence by ELISA and 40 herds(24.7%) by immunoblotting. The geographic distribution of herds with paratuberculosis was not uniform. Of the 241 feces samples including 110 feces from ELISA positive cow, 9 feces were positive in culture and PCR. PCR was able to detect the growth of M. paratuberculosis as early as 4 weeks of culture.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.4
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• pp.275-279
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• 2016

Dengue fever is a vector borne disease caused by a dengue virus. It is an RNA virus of the family flaviviridae, with different serotypes. Herein, we report our attempt to carry out a sensitivity comparison of immunodiagnostic assays for dengue fever in dengue positive patients. Blood samples from 189 volunteers were collected. To determine the sensitivity of the NS1 test, two different types of tests-immunochromatographic tri-line test and rapid dengue test (RDT)-as well as IgM and IgG capture ELISA were performed. The result of RDT has shown that 59.7% of volunteers were IgM positive and 50.2% were IgG positive. Conversely, the results from capture ELISA shows 79.8% and 59.7% for IgM and IgG, respectively. The sensitivity of the capture ELISA test for IgM and IgG was higher than that of immunochromatographic tri-line rapid test, but the specificity was lower. Therefore, to confirm dengue fever, we recommend performing more detailed, investigative tests since a single test may not be sufficient.

• Korean Journal of Veterinary Research
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• v.48 no.4
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• pp.457-462
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• 2008

The objective of this study was to determine appropriate sample size that simulated different assumptions for diagnostic test characteristics and true prevalences when designing serological surveillance plan for pullorum disease and fowl typhoid in domestic poultry production. The number of flocks and total number of chickens to be sampled was obtained to provide 95% confidence of detecting at least one infected flock, taking imperfect diagnostic tests into account. Due to lack of reliable data, within infected flock prevalence (WFP) was assumed to follow minimum 1%, most likely 5% and maximum 9% and true flock prevalence of 0.1%, 0.5% and 1% in order. Sensitivity were modeled using the Pert distribution: minimum 75%, most likely 80% and maximum 90% for plate agglutination test and 80%, 85%, and 90% for ELISA test. Similarly, the specificity was modeled 85%, 90%, 95% for plate agglutination test and 90%, 95%, 99% for ELISA test. In accordance with the current regulation, flock-level test characteristics calculated assuming that 30 samples are taken from per flock. The model showed that the current 112,000 annual number of testing plan which is based on random selection of flocks is far beyond the sample size estimated in this study. The sample size was further reduced with increased sensitivity and specificity of the test and decreased WFP. The effect of increasing samples per flock on total sample size to be sampled and optimal combination of sensitivity and specificity of the test for the purpose of the surveillance is discussed regarding cost.

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