• Parasites, Hosts and Diseases
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• v.37 no.3
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• pp.171-179
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• 1999

Non-biting midges are known to contain potent inhalant allergens. IgE antibody responses to the crude extract of Chironomus kiiensis adults, a dominant chironomid species in Korea, were examined. With the IgE-ELISA or passive cutaneous anaphylaxis reactions, increased levels of chironomid-specific IgE were detected in the skin test positived human sera, or immunized BALB/c mouse sera with the crude extract adsorbed to alum. IgE-immunoblot analysis showed mafor IgE-reacting protein band patterns, which reacted with more than 50% of the skin test positive human sera, at 110, 80, 46, 40, 37, 34 and 31 kDa. The reactive band patterns were larely similar between skin test positive humans and immune BALB/c mice. However, the bands of 55, 31, 27, 26, 24 and 23 kDa were found only in sensitized humans, but not in immunized mice.

• Journal of Life Science
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• v.9 no.5
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• pp.556-563
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• 1999

Presence of antibody to the capsid protein p24 is the main diagnostic criterion, since this reflects reliable antibody response to HIV infection. However, it takes about 6-8 weeks for antibody production after infection and people who are infected but antibodies are not produced yet are classified as seronegative. Therefore, there is a strong need for an improved diagnostic method for better health security. As a first step for developing such an improved diagnostic system, gag protein of human immunodificiency virus type 1 was expressed in E. coli DH5$\alpha$. The gag fragment of HIV-1 (including a portion of p17 and whole p24) was amplified by polymerase chain reaction (PCR) and BamH I/EcoR I sites were created during PCR. The amplified DNA fragment was cleaved with BamH I/EcoR I and was subcloned into the GEX-2T vector which had been digested with BamHI/EcoRI, resulting gene fusion with gst gene of pGEX-2T. The recombinant DNA was transferred into E. coli DH5$\alpha$. The transformed bacteria were grown at 37$^{\circ}C$ for 3h and protein expression was induced with 0.1mM IPTG at $25^{\circ}C$ for 3h. Recombinant gag protein or GST-gag fusion protein was purified with glutathione-sepharose 4B bead and migrated as a single band when analyzed by 10% polyacrylamide gel. These proteins were confirmed by immunoblotting with anti-GST goat sera or Korean AIDS patients sera. The results of this study establish the expression and single step pulification of HIV-1 gag protein which can specifically bind with Korean AIDS patients sera.

• Biomolecules & Therapeutics
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• v.7 no.1
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• pp.14-21
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• 1999

The immunogenicity of the recombinant human basic fibroblast growth factor (rh-bFGF) was investigated by tests for active systemic anaphylaxis (ASA), passive cutaneous anaphylaxis (PCA), passive hemagglutination (PHA) and guinea pig maximization test (GPMT) in mice or guinea pigs. Guinea pigs were sensitized with rh-bFGF ($100-1000\;\mu\textrm{g}/kg$) or rh-bFGF-CFA mixture ($1000\;\mu\textrm{g}/kg$). All animals sensitized with rh-bFGF alone or mixture with CFA showed symptoms of anaphylactic shock. IgE antibodies to rh-bFGF were detected in sera obtained from rh-bFGF and rh-bFGF-Alum ($1000\;\mu\textrm{g}/kg$) sensitized mice, indicating that rh-bFGF has immunogenicity eliciting potential. IgG and/or IgM antibodies to rh-bFGF were also detected in all the sera obtained from sensitized mice by PHA. In the GPMT for delayed type skin reaction, no skin reaction was observed in sensitized guinea pigs after intradermal injection and dermal application of 0.01% rh- bFGF. However, these positive reactions were consistent with the results of another rh-bFGF, showing that rh- bFGF is a heterogenous protein to rodents. Considering the fact that rh-bFGF is a genuine human protein of which structure is identical to the endogenous human bFGF, it is thought that rh-bFGF is rarely associated with immunological problems in clinical use.

• Clinical and Experimental Reproductive Medicine
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• v.12 no.1
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• pp.41-46
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• 1985

Various immunoserologic and cellular immunity techniques have been used to explore the presence of antisperm antibodies in the serum and seminal plasma of male patients and in the blood and genital fluid of infertile women. Several recent comparative investigations using various assays to detect and quantitate levels of antibody to human spermatozoa have produced widely varying results. So the first WHO workshop on iso- and autonatibodies to human spermatozoa in 1974 tried to establish some unification in the techniques used. The purpose of this study is to compare the results of two methods-the Kibrick macro-agglutination test and the Isojima micro-immobilization test-using the same test materials based on recommandation from WHO workshop. The results are as follows: 1. Twenty normal controls showed negative reactions in all the 2 tests. Out of 25 patients, the positive sera were noted in 15 (60%) on the Kibrick test and 13 (51%) on the Isojima test. 2. Twelve (48%) out of 25 patients showed positive reactions in the two tests, and 16 (64%) out of 25 patients showed positive reaction in one or more tests. 3. The titers of the antisperm antibodies on the Kibrick test was higher than that on the Isojima test. Therefore, it seems to be possible to increase the chances of detection of the antisperm antibodies, if two tests are imployed.

• The Korean Journal of Nuclear Medicine
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• v.19 no.1
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• pp.95-102
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• 1985

The activation of adenylate cyclase of human thyrocytes in primary cell culture and the release of c-AMP into the medium are used to detect b-TSH and TSAb in sera of patients with autoimmune thyroid disease. Sera of patients are used directly as a part of cell culture without immunoglobulin precipitation. In the above TSI bioassay, TSAb pooled serum show c-AMP concentration between that of 1mU/ml and 10 mU/ml b-TSH but normal control pooled serum doesn't show any detectable c-AMP response. Ninety fiye percent of untreated Graves' patients shows TSAb activity above normal range, 20% of Hashimoto's and 36% of euthyroid Graves' patients show detectable TSAb activity.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.3
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• pp.1635-1642
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• 2013

Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (${\geq}$ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culture-positive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by mass spectrometry. The relative molecular weights and the identities of the proteins, based on MALDI TOF/TOF, were as follows: CagI (25 kDa), urease G accessory protein (25 kDa), UreB (63 kDa) and proline/pyrroline-5-carboxylate dehydrogenase (118 KDa). These identified proteins, singly and/or in combinations, may be useful for diagnosis of H. pylori infection in patients.

• Parasites, Hosts and Diseases
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• v.50 no.3
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• pp.233-238
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• 2012

The precise diagnosis of the acute toxoplasmosis in pregnant women and immunocompromsied patients has critical importance. Most of the commercially available assays use the whole Toxoplasma soluble extract as the antigen. However, the assays currently available for the detection of specific anti-Toxoplasma antibodies may vary in their abilities to detect serum immunoglobulins, due to the lack of a purified standardized antigen. The aim of this study was production and evaluation of the usefulness of the recombinant Toxoplasma gondii GRA7 antigen for the serodiagnosis of Toxoplasma gondii IgM and IgG by ELISA. A total of 70 T. gondii IgM positive sera, 74 T. gondii IgG positive sera, and 60 sera from subjects who were not infected with T. gondii were examined. These sera were shown different absorbance values in ELISA test. To control the specificity of the rGRA7 other parasitic diseases, for example, echinococcosis, malaria, leishmaniasis, fascioliasis, and strongyloidiasis were tested of which none showed positive results. Sensitivity and specificity of the generated recombinant IgG ELISA in comparison with commercial ELISA (com ELISA) were 89% and 90%, and the sensitivity and specificity of the generated recombinant IgM ELISA were 96% and 90%, respectively. The results obtained here show that this antigen is useful for diagnostic purposes.

• Korean Journal of Veterinary Service
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• v.34 no.3
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• pp.195-200
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• 2011

Swine influenza is an acute respiratory disease prevalent in pig-growing areas all around the world and plays the roles of an intermediate host to be transmitted to mammals including human beings through a genetic recombination with the avian influenza virus. Recognizing that people could be contracted with swine influenza, this study set out to investigate the seroprevalence of individual and multiple infections with two subtypes (H1N1 and H3N2) of the swine influenza virus in pig farms in the Gyeongnam region according to age, area, and season, as well as to provide basic data for the prevention and control of swine influenza. Used in the study were total 904 swine sera that were not vaccinated against the influenza gathered from the pig farms in the Gyeongnam region from November, 2009 to October, 2010. HerdChek SIV (H1N1, H3N2) ELISA kit (IDEXX Laboratories, USA) was used for antibody testing against swine influenza. The test results show that 370 sera (40.9%) were infected with either H1N1 or H3N2 with 37.3% (337 sera) being contracted with H1N1, 13.1% (118 sera) with H3N2, and 9.4% (85) with both H1N1 and H3N2.

• Journal of Korean Medical Science
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• v.33 no.51
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• pp.340.1-340.14
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• 2018

Background: Various pneumococcal vaccines have been evaluated for immunogenicity by opsonophagocytic assay (OPA). A multiplexed OPA (MOPA) for 13 pneumococcal serotypes was developed by Nahm and Burton, and expanded to 26 serotypes in 2012. The development of new conjugate vaccines with increased valence has necessitated expanded MOPAs to include these additional serotypes. In this study, we validated this expanded MOPA platform and applied to measure antibodies against 11 additional serotypes (2, 8, 9N, 10A, 11A, 12F, 15B, 17F, 20B, 22F, and 33F) in human sera. Methods: All materials, including serum, complement, bacterial master stocks, and HL-60 cells, were evaluated for assay optimization. Following optimization, the assay was validated for accuracy, specificity, and intra- and inter-assay precision with sera from adult donors following standard protocols. The assay was applied to evaluate functional antibodies of 42 sera immunized with 23-valent pneumococcal polysaccharide vaccine (PPV23). Results: The expanded MOPA platform was specific for all serotypes, with the exception of serotype 20. The assay results were highly correlated with those obtained from single-serotype OPA, indicating acceptable accuracy. The coefficients of variation were 7%-24% and 13%-39% in tests of intra- and inter-assay precision, respectively, using three quality-control samples. A MOPA that included 11 additional serotypes in the PPV23 was established and validated with respect to accuracy, specificity, and precision. The opsonic indices of immune sera were obtained using this validated assay. Conclusion: The expanded MOPA will be useful for evaluation of the immunogenicity of PPV23 and future conjugate vaccine formulations.

• Parasites, Hosts and Diseases
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• v.56 no.1
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• pp.71-74
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• 2018

Soluble antigens from an axenic culture of Entamoeba histolytica were used to develop a commercial ELISA kit to quantify anti-E. histolytica antibodies in sera of patients with extraintestinal amebiasis in non-endemic settings. The diagnostic specificity and sensitivity of the test were assessed retrospectively using 131 human serum samples with amoebic serologic status available. They were selected according to their results in immunofluorescence (IFAT) and were separated in 2 sample categories: 64 sera with positive results by IFAT and 67 with negative results by IFAT. The sensitivity and specificity of the ELISA kit were assessed at 95.0% and 94.0% compared to the IFAT. The test can be useful to exclude a potential diagnosis of amebiasis and could be used as a screening method since ELISA is an automated technique.