• 대한수의학회지
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• 제38권1호
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• pp.53-63
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• 1998

Total of 1085 swine sera (1996-1997) from nation-wide were tested for the presence of antibodies to influenza A virus. Fifty nine percent of the tested sera showed seropositive by HI test. Positive sera consisted of 24--- of H3, 15--- of H1, and 20--- of the sample had both antibodies, respectively. Sera collected from various region represented 7~27--- seropositivity to H1N1, 15~25--- to H3N2, respectively. Swine influenza field isolate from nasal swab was characterized antigenically and genetically to elucidate its relatedness with other known strains of influenza A virus. The study was focused on the HA gene which is related to pathogenecity and antigenic variability of the influenza virus. By RT-PCR using influenza A/H1N1 specific primers, influenza virus H1N1 specific DNA fragment was amplified from A/Swine/Iowa/15/30(H1N1), US field isolate but not in H3N2 strain. PCR products were sequenced by dideoxy chain termination method to determine nucleotide homology with other strains of influenza A virus. The US field isolate and A/Swine/Indiana/1726/88 strain had 97--- of nucleotide homology and 98--- of amino acid homology. Based on the results obtained from this experiment, the field isolate was genetically related to A/Swine/Indiana/1726/88 and had higher homology with A/Swine/Indiana/1726/88 than with classical swine influenza virus, A/Swine/Iowa/15/30. The field isolate had no amino acid changes at the antigenic site compare to that of the A/Swine/Indiana/1726/88. The proteolytic enzyme cleavage site between HA1 and HA2 had no alteration and the amino acid arginine was intact. There is no evidence has been found that the field isolate has genetic shift or genetic drift which might altered antigenic determinant.

• 한국동물위생학회지
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• 제34권3호
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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.

• Archives of Pharmacal Research
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• 제24권3호
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• pp.256-261
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• 2001

Allergenicity of genetically-modified (GM) soybean was evaluated in male Sprague Dawley rats. To confirm the GM soybean used in this study, the polymerase chain reaction (PCR) was performed using the chromosomal DNA of soybeans. The PCR result provided the clear discrimination of genetically-modified (GM) soybeans. To evaluate the allergenicity of GM soybean and non-GM control one, the soybean homogenate was sensitized subcutaneously 3 times a week for 3 weeks. The doses of soybean were 0, 2 and 20 mg/kg in the protein basis. A week after the last sensitization, antisera were recovered from individual animals. When the sera were injected intradermally on the clipped back of unsensitized rats with various dilutions, followed by a challenge with 20 mg/kg of soybean homogenate containing 1% Evans blue, no sign of passive cutaneous anaphylaxis reaction was detected. In addition, when the sera were treated in the cultures of peritoneal mast cells, the increase of histamine release by anti-(GM soybean) sera was not observed when compared to that by anti-(non-GM soybean) sera. The present results indicate that the GM soybean might not act as a strong allergen in male Sprague Dawley rats.

• Parasites, Hosts and Diseases
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• 제41권4호
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• pp.203-207
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• 2003

An ELISA was developed for the diagnosis of vivax malaria using multiple stage-specific recombinant antigens of Plasmodium vivax. The DNA from the whole blood of a malaria patient was used as template to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1). Each amplified DNA fragment was inserted into pQE30 plasmid to induce the expression of His-tagged protein in Escherichia coli (M15 strain) by IPTG. His-tagged proteins were purified by Ni-NTA metal-affinity chromatography and used as antigens for ELISA with patient sera that were confirmed previously by blood smear examinations. When applied to patient sera, 122 (80.3%) out of 152 vivax malaria cases reacted to at least one antigen, while no reactions were observed with 128 uninfected serum samples. We applied this ELISA to the screening of 3,262 civilian residents in endemic regions near the DMZ, which resulted in 236 positively detected (7.2%) cases. This method can be applied to serological diagnosis and mass screening in endemic regions, or can be used as a safety test for transfusion blood in endemic areas.

• Journal of Korean Medical Science
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• 제33권51호
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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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• 제60권1호
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• pp.25-34
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• 2022

Alveolar echinococcosis (AE) caused by infection with E. multilocularis metacestode, represents one of the most fatal helminthic diseases. AE is principally manifested with infiltrative, proliferating hepatic mass, resembling primary hepatocellular carcinoma. Sometimes metastatic lesions are found in nearby or remote tissue. AE diagnosis largely depends on imaging studies, but atypical findings of imaging features frequently require differential diagnosis from other hepatic lesions. Serological tests may provide further evidence, while obtaining reliable AE materials is not easy. In this study, alternative antigens, specific to AE were identified by analyzing E. granulosus protoscolex proteins. An immunoblot analysis of E. granulosus protoscolex showed that a group of low-molecular-weight proteins in the range from 14 kDa to 16 kDa exhibited a sensitive and specific immune response to AE patient sera. Partial purification and proteomic analysis indicated that this protein group contained myosin, tubulin polymerization promoting protein, fatty-acid binding protein, uncharacterized DM9, heat shock protein 90 cochaperone tebp P-23, and antigen S. When the serological applicability of recombinant forms of these proteins was assessed using enzyme-linked immunosorbent assay, DM9 protein (rEgDM9) showed 90.1% sensitivity (73/81 sera tested) and 94.5% specificity (172/181 sera tested), respectively. rEgDM9 showed weak cross-reactions with patient sera from the transitional and chronic stages of cystic echinococcosis (3 to 5 stages). rEgDM9 would serve as a useful alternative antigen for serodiagnosis of both early- and advanced-stage AE cases.

• 대한수의학회지
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• 제63권2호
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• pp.12.1-12.7
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• 2023

Bovine viral diarrhea virus (BVDV) is an RNA virus belonging to Pestivirus in the family Flaviviridae. BVDV has economic significance for the livestock industry because of its association with acute disease, fetal loss, and birth of persistently infected (PI) animals. This study aimed to investigate the BVDV infection rates in Korean native cattle farms in Jeju for further planning of a BVDV control program in the Jeju Province. BVDV antibodies and antigens were tested in 15,842 sera collected from 302 Korean native cattle herds between January 2014 and June 2017 using enzyme-linked immunosorbent assay (ELISA). Viral antigen was detected by reverse transcription-polymerase chain reaction from 60 sera that were antigen ELISA-positive. BVDV antibodies were found in 90.7% (274/302) herds and 61.1% (9,678/15,842) cows. BVDV antigens were found in 13.2% (40/302) herds and 0.4% (61/15,842) cows. The oldest animal group (> 8 years) exhibited the highest sero-positive rates (91%), while the youngest animal group (< 1 years) had the highest antigen positivity rates (0.52%). Of the 60 antigen-positive sera, BVDV types 1 and 2 were found in 36 and 12 sera, respectively. Additionally, six animals were considered to be PI as BVDV was continually detected in annual examination.

• 대한미생물학회지
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• 제20권1호
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• pp.79-89
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• 1985

It is important to discriminate between tuberculosis and tuberculosis-like disease by Mycobacteria other than tuberculosis in the serodiagnosis of tuberculosis. But because common antigens share among Mycobacteria, their antigenicities to human are similar. Therefore degree of cross-reactivity of antibody in the sera of patients with tuberculosis between M. tuberculosis and Mycobacteria other than tuberculosis should be checked to increase the specificity in the serodiagnosis of tuberculosis. The activity levels of IgG antibody in the sera of 106 patients confirmed as active pulmonary tuberculosis and 30 normal healthy control person to the pressate extract antigen (TE, BE, AE, and FE antigen) from M. tuberculosis, M. bovis, M. avium, and M. fortuitum were measured by enzyme-linked immunosorbent assay and the crossreactivity of IgG antibody with mycobacterial species was analysed. The results were as follows; 1. The activity level(O.D. at 492nm) of IgG to TE antigen in sera of patients with pulmonary tuberculosis was $0.228{\pm}0.167$ in minimal tuberculosis; moderately advanced, $0.556{\pm}0.616$; far advanced, $1.116{\pm}0.651$ and $0.315{\pm}0.245$ in miliary tuberculosis. 2. The activity level (O.D. at 492nm) of IgG to BE antigen in sera of patients with pulmonary tuberculosis was $0.190{\pm}0.162$ in minimal tuberculosis; moderately advanced, $0.337{\pm}0.361$; far advanced, $0.713[\pm}0.460$ and $0.204{\pm}0.162$ in miliary tuberculosis. 3. The activity level (O.D. at 492nm) of IgG to AE antigen in sera of patients with pulmonary tuberculosis was $0.165{\pm}0.114$ in minimal tuberculosis; moderately advanced, $0.392{\pm}0.494$; far advenced, $0.751{\pm}0.512$ and $0.233{\pm}0.191$ in miliary tuberculosis. 4. The activity level (O.D. at 492nm) of IgG to FE antigen in sera of patients with pulmonary tuberculosis was $0.280{\pm}0.227$ in minimal tuberculosis; moderately advanced, $0.460{\pm}0.564$ ; far advanced, $0.845{\pm}0.573$ and $0.257{\pm}0.103$ in miliary tuberculosis. 5. The activity level (O.D. at 492nm) of IgG in sera of healthy control person was $0.126{\pm}0.084$ to TE antigen. $0.105{\pm}0.041$ to BE antigen, $0.103{\pm}0.052$ to AE antigen, and $0.095{\pm}0.061$ to FE antigen. 6. Degree of correlation(r) in activity level of IgG between TE antigen and BE antigen was 0.905 ; between TE antigen and AE antigen, 0.760; between TE antigen and FE antigen, 0.790, and between AE antigen and FE antigen, 0.945. 7. As O.D. above 0.200 was determined positive for the serodiagnosis of pulmonary tuberculosis, the sensitivity and specificity in ELISA using TE antigen were 80% and 87% respectively, whereas in the case of using BE antigen, 66% and 100%; in the case of using AE antigen, 62% and 100%, and in the case of using FE antigen, 72% and 93%, respecitively.

• 대한미생물학회지
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• 제18권1호
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• pp.73-85
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• 1983

The advantages of the enzyme-linked immunosorbent assay(ELISA) are its sensitivity and its simplicity in detecting IgM and IgG antibodies. For applying the ELISA to the diagnosis of typhoid fever, first of all, experiments were performed to determine which concentration of killed whole cell antigens and lipopolysaccharide(LPS) antigens of S. typhi(0.901 w) were optimally coated to the wells of the polystyrene and polyvinylchloride microplate, using the hyperimmune sera from rabbits against S. typhi. By using both kinds of antigens of S. typhi adsorbed to the ELISA microplate, the changing patterns of IgG and IgM antibodies in the sera from rabbits responding to the killed whole cell antigens of S. typhi(0901 w) during the prolonged immunization were serially traced by the ELISA. At the same time, the level of antibodies against S. typhi in sera fron patients with typhoid fever and from normal healthy persons were measured by the ELISA employing the killed whole cell antigens and LPS antigens as the coating antigens. The results obtained were summerized as follow: 1. The optimal concentration of the killed whole cell antigens, which were more easily adsorbed to the polystyrene plate than the polyvinylchloride plate, was $10^8cells/ml$ of carbonate buffer(pH. 9.6) on the wells of the polystyrene plate when treated at $37^{\circ}C$ for 4 hours. On the other hand, the optimal concentration of lipopolysaccharide antigens, which were adsorbed only to the polyvinylchloride plate, was $100{\mu}g/ml$ of carbonate buffer(pH. 9.6) on the wells of the polyvinylchloride plate when treated at $37^{\circ}C$ for 4 hours. 2. IgM antibody response were dominating in rabbits responding to the killed whole cell antigens of S. typhi(0.901 w), and were more specific to the LPS antigens than to the killed whole cell antigens in the ELISA. Good correlations were made between the IgM titers by the ELISA and the aggglutinating titers of sera from the immunized rabbits. 3. Both IgG and IgM agglutination titers by the ELISA in sera from most of patients with typhoid fever were above 1:320 but those in sera from most of normal, healthy persons were below 1:80. 4. There were close correlations between the antibody titers by the ELISA and the agglutinating titers to the killed whole cell antigens in the tested human sera, IgM titers being more correlated with the agglutinating titers than IgG titers. But a little correlations were made between the antibody titers by the ELISA and the agglutinating titers to the LPS antigens. 5. IgM titers in the tested human sera were similar to IgG titers detected by the ELISA employing the killd whole cells antigens and the LPS antigens. 6. Good correlations were made between the antibody titers demonstrated by the ELISA performed on the killed whole cell antigens and the LPS antigens as the different, coating antigens on the ELISA microplates.

• Parasites, Hosts and Diseases
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• 제21권1호
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• pp.1-5
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• 1983

간츱충증 환자로부터 혈액을 채취하여 그 혈청에서 간흡충에 대한 항체를 Thin Layer Immunoassay(TIA)에 의하여 검출하였다. TIA는 Elwing et al.(1976)이 기술한 방법에 의하여 시행하였고, 항원은 이등(1981)의 방법으로 제조된 간흡충 성충 생리식염수 추출물을 사용하였으며 그 단백질 함량은 2.87mg/ml이었다. 간흡충중 환자 60명의 혈청에서의 항체가는 건강대조군 및 이질아메바 감염자에서의 항체가와 비교할 때 그 차이가 유의하였으나, 폐흡충증 환자와의 차이는 유의하지 않았다. 간흡충중 환자의 성별이나 연령과 TIA에 의한 항체가 사이에 상관 관계가 없었다. 환자의 분변내 EPG, 말초혈액내 호산구의 수와도 상관이 없었다. TIA에 의한 항체가와 간기능 검사 결과와의 관계를 보면 alkaline phosphatase, SGOT 및 SGPT 측정치와 모두 상관이 없음을 알 수 있었다.