• 한국동물위생학회지
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• 제17권2호
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• pp.95-113
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• 1994

To establish an effective diagnostic measure for detection of the antibodies against Actinobacillus pleuropneumoniae, the methods for tube agglutination test (TAT), plate agglutination test (PAT), micro-agglutination test(MAT) and agar-gel immunodiffusion test(ID) were improved and standarized, and the comparative studies were carried out. The results obtained through the experiments were summarized as follows. 1. The rabbit hyperimmune sera to reference serotypes 1 to 6 were cross-tested with TAT, PAT, MAT and ID. In the homologous systems, the range of antibody titers in TAT was 80 to 640, showing the cross-reaction in serotypes 3, 4, 5 and 6. The range of antibody titers in PAT was 4 to 64, showing the cross-reaction in serotypes 3, 4, 5 and 6. In ID, the range of antigen titers was 8 to 32, and cross-reaction was observed in serotype 5. 2. The optimal concentration of antigen in PAT and MAT were 100mg /ml and 1.25mg /ml respectively. The most sensitive reaction in MAT was observed in 52$^{\circ}C$ for 18hrs. 3. In ID, the most promising antigen and the buffer for agar-gel were EDTA-treated antigen and 0.05M tris buffer (pH 7.2), respectively. 4. By the tests for 200 swine sera, it was found that the frequency of positive reaction were 203 in TAT, 240 in PAT and 163 in ID. 5. When compared the titers of TAT with those of MAT for 200 swine sera, MAT showed the higher titer than TAT being increased by relative correlation. Int was found that the titer for positive readings were 20 in TAT and 40 in MAT. 6. when compared the results of ID with those of TAT for 200 swine sera, all sera with TAT titer under 10 were negative in ID. Of the sera with TAT titer 20 and 40, 55.1% nd 91.8% were positive in ID, respectively. All sera with TAT titer above 80 were positive in ID. In comparison of ID and MAT, all sera with MAT titer under 20 were negative in ID. Of the sera with MAT titer 40 and 80, 24.7% and 93.9% were positive in ID, respectively. All sera with MAT titer over 160 showed positive in ID. 7. In conclusion, the established MAT showed high sensitivity but low specificity, wherease ID revealed low sensitivity but high specificity.

• Parasites, Hosts and Diseases
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• 제36권4호
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• pp.269-275
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• 1998

톡소포자충과 네오포자충 (Neospora caninum)의 충체 항원으로 톡소포자충 양성 혈청 및 톡소포자충증 환자의 혈청과 ELISA, western blot 및 면역형광법을 실시하였다. ELISA에서는 172명의 톡소포자충 양성 혈청에서 12명 （6.7％）이 두 항원에 모두 반응하였으며, 톡소포자충 음성인 110명의 혈청에서 1명이 네오포자충과 반응하였다. 교차반응을 보인 12 명의 혈청을 western blot으로 확인하였을 때, 톡소포자충 항원과는 다양한 양상으로 반응하였으나, 30 kDa (SAG1)과 22 kDa (SAG2) 항원과 강하게 반응하였다. 네오포자충과의 반응은 급격히 감소하였으나, 세 경우에서 43 kDa 단백질과 반응하였으며, 음성 혈청군의 1명도 43 kDa 단백질과 반응하였다. 면역형광법에서는 모든 양성 혈청이 톡소포자충의 세포막에 표지되었으나, 네오포자충과의 반응은 세포막, 세포 내 소기관, 혹은 둘 다를 표지하였다. 이로써, 톡소포자충과 네오포자충의 항원적 교차반응과 사람 혈청에서 네오포자충에 대한 항체의 존재를 확인하였으며, 네오포자충에 의한 인체감염 가능성에 대해서는 추후 연구가 필요할 것으로 판단된다. 또, N. caninum의 우리말 이름을 네오포자충으로 제안한다.

• Parasites, Hosts and Diseases
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• 제26권4호
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• pp.245-254
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• 1988

유구낭미충(이하 낭미충)(Cysticercus cellulosae)에서 추출한 조항원으로 낭미충증(cysticercosis), 스파르가눔중(sparganosis) 및 포충중(hydatidosis) 환자 및 정상인 혈청내 IgG 항체와의 혈청학적 반응을 ELISA 및 EITB 방법으로 추적하였다. ELISA에서 낭미충의 조항원 단백은 스파르가눔중 환자 혈청과의 교차 반응도 보였고, 포충증 환자 혈청에 대해서는 낭미충 환자 혈청보다 오히려 높은 흡광도(O.D.)를 보였다. 낭미충 조항원을 SDS-PAGE로 전기영동한 결과 260 KDa∼22 KDa 범위에서 31개의 polypeptide band를 얻었으며 이 중 11개의 분획이 강한 염색성을 나타내었다. 낭미충 조항원과 낭미충중 환자 혈청과의 EITB 반응에서 22개의 항원대가 관찰되었으며 이 중 104 KDa, 82 KDa, 78 KDa 및 59 KDa는 모든 검사 혈청에서 인지되었다. 또 22개의 항원대 중 12개는 정상인 혈청에서도 반응이 관찰되어 교차반응하는 비특이성 항원 성분으로 해석되었다. 낭미충 조항원과 스파르가눔증 환자 혈청과의 반응에서 11개의 항원대가 인지되었으며 이 중 낭미충증 환자 혈청과 교차반응하는 9개의 항원대가 인지되었다. 나머지 163 KDa 및 131 KDa 항원대는 스파르가눔 환자에서만 동정되었다. 낭미충 조항원과 포충증 환자 혈청과의 반응에서는 21개의 항원대가 인지되었는데, 이 중 63 KDa는 포충증 환자에서만 인지되었고 나머지는 낭미충증 환자 혈청과 교차반응하는 항원대이었다. 한편 낭미충 조항원 중 104 KDa, 82 KDa, 72 KDa, 59 KDa및 34 KDa는 낭미충증, 스파르가눔증 및 포충중 환자와 정상인의 혈청에서도 모두 교차반응을 일으키는 항원이었다.

• 한국동물위생학회지
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• 제15권2호
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• pp.174-183
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• 1992

This study was conducted to determine the serum antibodies against toxoplasma in swine from breeding-pig farm, pig farm and abattoir by latex agglutination(LA) test. LA test was carried out with commercial Toxo-MT kit (Eiken chemical co.). The results obtained were summerized as follows : 1. The cut-off titer of positive and negative reactions by Toxo-MT antigen used in this experiment was determined as the serum dilution of 1 ; 32. 2. positive rates of toxoplasma antibodies in 823 swine sera were 17.0%(140 cases) by LA test. 3. The toxoplasma antibody detection rates against 194 swine sera in breeding-pig farm, 273 swine sera in pig farm and 356 swine in abattoir were 46.9%(91 cases), 8.4%(23 cases) and 7.3% (26 cases) , respectively. 4. In LA test serum antibody titers in 823 test sera were shown as 51 cases (36.4%) in 1 : 32, 40(28.6%) in 1；64, 17(12.1%) in 1:128, 14(10.0%) in 1：256, 10(7.1%) in 1:512, 5(3.6%) in 1：1,024, and 3(2.1%) in 1 : 2,048. 5. Positive rates of toxoplasma antibodies in swine sera from each breeding-pig farm were 20.0∼61.9%.

• 대한미생물학회지
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• 제21권2호
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• pp.205-210
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• 1986

Serological investigations for the leptospirosis on hospitalized patients in Choonchun Sungsim Hospital during the periods from August to November 1985 and 841 inhabitants of Kangwondo area including Choonchun, Choonsung, Inje, Chulwon, Hwachun, Gosung, Taibaik, Samchuk and Yangju area were carried out. 1. Among 58 hospitalized patients who were suspected as leptospirosis, 10 patients were detected to have antibody against Leptospira. All of positive sera had the highest antibody titer against serogroup Icterohemorrhagiae and most positive sera were also reactive to serogroup Australis and Canicola. Antibody titer of positive sera detected by microscopic agglutination(MA) test were ranging from 1 : 40 to 1 : 2,560. Antibody titer detected by ELISA method were higher than those detected by MAT(ELISA 1 : 400$\sim$1 : 25,600) and IgM titer of positive sera were generally higher than IgG titer. 2, Of 841 inhabitants in 8 area of Kangwondo, 17 persons (2,02%) possessing antibody against Leptospira were detected by ELISA method, IgG titer in positive sera were generally higher than IgM titer. Persons possessing antibody to Leptospira were distributed in both sex and in various age group, and no significant regional and occupational fluctuations were obserbed.

• Parasites, Hosts and Diseases
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• 제40권4호
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• pp.195-198
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• 2002

This study was performed to investigate whethere there is cross-reactivity between Dirofilaria immitis and three intestinal nematodes of dogs. In ELISA, D. immitis- infected dog sera obtained at the 4th molting stage (9-11 weeks) and microfilaremic stage (25-30 weeks) were shown to be highly reactive with crude extract of T. canis. In immunoblotting, some antigenic fractions, 44, 57 88, 100 kDa of crude extract of T. canis, were found to be positive reaction with sera of dogs infected with D. immitis. However, little or no cross-reaction were observed between sera of D. immitis-infected dogs and crude extract antigen of T. vulpis or A. caninum. These result suggest that there are partial cross reaction between sera of D. immitis-infected dogs and the antigen of T. canis.

• 한국동물위생학회지
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• 제22권4호
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• pp.385-392
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• 1999

A total of 420 chicken sera from various regions were tested for the presence of antibodies to avian rotavirus using indirect immunofluorescence assay (IFA). In broiler farms, rotavirus antibodies were detected from 20 farms among 30 farms tested and the positive rates were above 50% in 9 farms. In parent stock farms, rotavirus antibodies were detected from 5 farms among 14 farms tested. From sera collected in 7 layer farms rotavirus antibodies were not detected.

• Parasites, Hosts and Diseases
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• 제47권1호
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• pp.73-77
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• 2009

Differential diagnosis of Taenia asiatica infection from other human taeniases by serology has been tested. An enzyme-linked immunoelectrotransfer blot (EITB) was applied to subjected human sera and tapeworm materials. Thirty-eight proteins reactive to serum IgG were observed between 121 and 10 kDa in adult worms, and more than 22 serum-reactive components between 97kDa and 21.5kDa were observed in eggs of T. asiatica. Antigens of adult T. asiatica revealed immunoblot bands between 120 and 21.5 kDa against T. asiatica infected sera. Antigens of adult Taenia saginata revealed 110-100, 66, 58-56, and 46 kDa immunoblot bands against T. asiatica infected sera. Antigens of adult Taenia solium also revealed 99-97, 68-66, and 46 kDa bands against T. asiatica infected sera. The immunoblot band of 21.5 kDa exhibited specificity to T. asiatica.

• BMB Reports
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• 제29권3호
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• pp.192-199
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• 1996

A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.

• Parasites, Hosts and Diseases
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• 제39권2호
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• pp.171-176
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• 2001

Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), meroxoite surface protein (MSP-1), apical merozoite antigen (AMA- 1), serine repeat antigen (SERA), and exported antigen (EXP- 1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using S antigens (91.9%) rather than using each antigen solely (55.6 - 80%), a combination of 2 (76.3 - 87.5%), 3 (85.6 - 90.6%), or 4 antigens (89.4 - 91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.