• 대한수의학회지
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• 제29권4호
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• pp.567-575
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• 1989

This study was conducted to evaluate the possibility of application of a microenzyme-linked immunosorbent assay(micro-ELISA) for the serodiagnosis of specific toxoplasma antibodies in swine sera and this test was performed as a microplate system by coating the polystyrene plates with toxoplasma soluble antigen, incubated serially diluted sera, then added horse radish peroxidase labelled goat anti-swine IgG(r) conjugate followed by o-phenylenediamine as substrate. The color development by enzyme-substrate reaction was determined by the photometric reading [ELISA reader at 490nm (OD)] and visual reading. The soluble antigen was prepared from the tachyzoites in mouse peritoneal cavity. A total of 1,200 swine sera from pig slaughter-house and a total of 116 swine sera from pig breeding station (S-C farm) were tested for the detection of antibodies to Toxoplasma gondii. The results obtained were summarized as follows: 1. The optimal reactions of indirect ELISA for the test sera were determined by the dilution of antigen 1:256 and 1:3,200 of horse radish peroxidase conjugate [anti-swine IgG(r)]. 2. The specific togoplasma antibody(IgG) in pigs infected with Tp artificially were detected as the serum titers of 1:64 or 1:128 at one week postinfection. 3. Of a total of 1,200 swine sera from pig slaughter-house 505 samples of sera were detected as positive (42.1%) and of a total of 116 swine sera from S-C pig breeding station 68 samples of sera as positive (58.6%). 4. The specific antibody(IgG) detection rates against a total of 1,200 test sera from pig slaughter-house were not significant between male (43.1%) and female (40.7%). 5. The indirect ELISA was proved to be a sensitive and specific procedure for the serodiagnosis of swine toxoplasmosis and also evaluated as an effective screening test for the large scale of test samples in laboratory.

• 미생물학회지
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• 제21권1호
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• pp.15-26
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• 1983

Commercial deerhorns were extracted in boiling water, the deerhorn extracts were per os introduced into rabbits, and then the effects of the extracts on the antibody productions aginst Escherichia coli antigens were investigated for 4 weeks. The experimental rabbits were divided into 4 groups ; control, only deerhorn, only antigen, and antigen plus deerhorn treated groups. The effects of the treatments were measured by counting the number of blood cells, weighing, and immunoelectrophoresis. The rabbits' body weights gained up to 185% in the deerhorn group, and the other groups gained nearly 120%. The numbers of red blood cells in the antigen plus deerhorn group increased somewhat. However, the numbers of leucocytes gradually increased after one week in the antigen group, and at 4 weeks increased up to 290%. In the antigen plus deerhorn group the numbers of leucocytes increased suddenly up to 189% at one week, but after one week the numbers recovered to normal state. Strangely in the deerhorn group the numbers decreased up to 40%. The amounts of serum globulin increased in the control after one week, but maintained about 130%. In the deerhorn group the amounts increased like the control, but after 4 weeks increased up to 175%. In the antigen group the amounts were not changed until 2 weeks, but after 3 weeks abruptly increased over 175%. In the deerhorn plus antigen group the amounts increased gradually up to 262% until 3 weeks, after 3 and 4 weeks the amounts did not increase. The amounts of serum .gamma.-globulin decreased in the control group, and in the deerhorn group and antigen group the amounts did not change until 2 weeks, but after 3 weeks abruptly increased over 175%. In the deerhorn plus antigen group the amounts increased gradually up to 262% until 3 weeks, after 3 and 4 weeks the amounts did not increase. The amounts of serum ${\gamma}-globulin$ decreased in the control group, and in the deerhorn group and antigen group the amounts did not change until 3 weeks, but after 4 week the amounts slightly increased up to 110%. However, the amounts in the deerhorn plus antigen group did not change until 2 weeks, but after 2 weeks abruptly increased up to 174%. The recognized immunoglobulins were IgG and IgM, and the enhanced immunoglobulin was IgG.

• 한국동물위생학회지
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• 제15권1호
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• pp.32-45
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• 1992

In order to establish and enzyme-linked immunosorbent assay to ILTV, field virus strain of ILTV was propagated in chorioallantoic membrane of the embryonated eggs. purified and used as antigen. The antisera selected from the field samples and immunized chickens based on serum neutralization test were used as the standard positive and negative sera in all tests. It was found that optimal antigen concentration was $2{\mu}g$ of protein per well and a 1 : 100 dilution of standard serum showed low background optical density with negative serum and high P/N values of positive sera. A 1 : 500 dilution of the rabbit anti-chicken IgG peroxidase conjugate produced a high P/N values and thirty minutes was chosen as suitable time to read the optical density of the enzyme substrate reaction and optical density was consistent during the 16 hours after stopper was treated. When coated antigen was kept on microplate for varying time up to 16 hours at $4^{\circ}C$ or $37^{\circ}C,$ no significant difference was observed between the treatment. The coated antigen could be kept without change of antigenicity for at least one month at $-70^{\circ}C,\; -20^{\circ}C,\; 4^{\circ}C$ and room temperature. When blocking buffer contanining bovine serum albumin was mixed directly with conjugate and serum at 10% level induced higher P/N values compared to blocking antigen coated microplate with the same blocking buffer. The coefficience of correlation between ELISA and SN test was 0.577. When antibody response of chickens, vaccinated with ILTV, was examined by ELISA and SN test, antibody rising and decay pattern between the two test was similar until 11 weeks of age. However 12 weeks onward antibody titer checked on by SN test was slightly lower than that tested by ELISA.

• Parasites, Hosts and Diseases
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• 제46권3호
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• pp.195-198
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• 2008

Plasmodium falciparum liver stage antigen-1 (PfLSA-1) is one of the few antigens expressed exclusively in liver stage parasites. In this study, we evaluated the antibody responses against recombinant PfLSA-1 in naturally infected individuals in Myanmar. High levels of antibody responses (70.7%) were detected in 82 serum samples from 116 infected individuals, and IgG responses to PfLSA-1 principally composed of responses of IgG1 and IgG3 subclasses. These results show that PfLSA-1 elicits effective antibody responses in individuals infected with P. falciparum, and thus it could be not only an attractive candidate protein for vaccine development, but also a useful antigen for serodiagnosis of the infection.

• 한방안이비인후피부과학회지
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• 제21권1호
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• pp.38-54
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• 2008

Objective : This study was designed to investigated effects of Pinelliae Rizoma (PR) on asthma Methods : Detecting antigen specific antibody isotypes, cytokines in serum and bronchoalveolar lavage fluid (BALF). In addition, the present author also investigated changes in weight of spleen and proliferation rates of splenocytes. Finally, histopathological observation of the lung tissue was also investigated. Results : Oral administration of PR lowered OVA specific IgE levels in serum, IgG1 levels in serum and BALF. PR also decreased production levels of IL-4 in BALF. In addition, total cells in BALF were decreased by oral administration of PR effectively. In histapathological observation, PR group showed downward tendency of inflammatory cell infiltration around small vessels. Conclusion : PR is useful to treat patients with asthma and the mechanisms are related in suppression of Th2 skewing reactions.

• Parasites, Hosts and Diseases
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• 제60권1호
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• pp.7-14
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• 2022

Acanthamoeba keratitis (AK) is a rare infectious disease and accurate diagnosis has remained arduous as clinical manifestations of AK were similar to keratitis of viral, bacterial, or fungal origins. In this study, we described the production of a polyclonal peptide antibody against the adenylyl cyclase-associated protein (ACAP) of A. castellanii, and evaluated its differential diagnostic potential. Enzyme-linked immunosorbent assay revealed high titers of A. castellanii-specific IgG and IgA antibodies being present in low dilutions of immunized rabbit serum. Western blot analysis revealed that the ACAP antibody specifically interacted with A. castellanii, while not interacting with human corneal epithelial (HCE) cells and other causes of keratitis such as Fusarium solani, Pseudomonas aeruginosa, and Staphylococcus aureus. Immunocytochemistry (ICC) results confirmed the specific detection of trophozoites and cysts of A. castellanii co-cultured with HCE cells. The ACAP antibody also specifically interacted with the trophozoites and cysts of 5 other Acanthamoeba species. These results indicate that the ACAP antibody of A. castellanii can specifically detect multiple AK-causing members belonging to the genus Acanthamoeba and may be useful for differentially diagnosing Acanthamoeba infections.

• 공업화학
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• 제30권4호
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• pp.429-434
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• 2019

본 논문에서는 수용성의 CdSe/ZnS 양자점을 합성하고 이에 항체기능성을 도입하여 lateral flow immunoassay (LFIA) 플랫폼에 융합하여 폐암 질병진단에 활용 가능한 단백질 바이오마커[예: 인간 혈청 아밀로이드 A-1 (hSAA1)]의 농도 분석에 적용하고자 한다. 면역분석법 센서 스트립은 니트로셀룰로오즈 막에 테스트라인과 대조라인으로 각각 항hSAA1 단일클론항체(10G1)(anti-hSAA1)와 항chicken IgY (anti-chicken IgY)를 스프레이하여 제작하였다. 이와 함께, 유기상에서 합성된 CdSe/ZnS 양자점은 카르복실기로 변형된 알케인티올기를 이용한 리간드 교환방법으로 수용성으로 전환하였으며, 이에 타겟 단백질인 hSAA1에 특이적으로 결합 가능한 항체인 항hSAA1 단일클론항체(14F8)로 컨쥬게이션하여 형광검출용 입자[QDs-anti hSAA1 (14F8)]로 사용하였다. 제작된 LFIA 스트립 위에 순차적으로 다른 농도의 hSAA1과 QDs-anti hSAA1 (14F8)의 복합체를 흘려주면, 테스트라인에 anti hSAA1 (10G1)/hSAA1/QDs-anti hSAA1 (14F8) 샌드위치 복합체가 형성되어 양자점에 의한 발광신호가 검출됨을 측정하였다. 최적화된 측방흐름이 가능한 완충용액 조건에서 100 nM 농도의 hSAA1 단백질의 유무를 5 min 안에 눈으로 확인 가능하였다.

• Journal of Nutrition and Health
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• 제29권9호
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• pp.971-980
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• 1996

The soluble transferrin receptor(TfR) in human serum has been shown recently to be a truncated form of intact membrane bound receptor containing most of the extracellular domain. We purfied the transferin-free TfR from human serum by immounoaffinity chromatography which produced the single protein identity in high resolution gel chormatography. The monoclonal antibodies(MAb) against purifed serum TfR were produced by fusion of spleen cells o fimmunized Balb/c mice and SP2 cells. Ten hybrids producing MAb specific for serum TfR were identifed and determine their iostypes. A immunoraddiometric assay (IRMA) for serum TfR was established using two monoclonal IgG1 antibodies as the coating and indicator antibodies on the bosis of their suitability in sandwich IRMA of serum TfR. The mean serum TfR levels in the 15 normal male, 15 normal female, and 19 iron-deficient subjects were 5.4$\pm$0.98, 4.6$\pm$0/76, and 18.0$\pm$12.8mg/1, respectively, and the difference in mean values between normal and iron deficient subjects was significant(p=0.0005). There existed the inverse logarithmic relationship(r=-0.9336, p<0.0001) between the serum TfR and ferritin levels.

• Parasites, Hosts and Diseases
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• 제27권4호
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• pp.261-270
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• 1989

스파르가눔증(sparganosis)은 Spirometra의 plerocercoid 유충이 인체에 감염되었을 때 유발되는 질병으로 국 내에서 드물지 않게 관찰되는 조직내 기생 윤충중이다. 이 연구에서는 마우스에 실험적으로 피하 스파르가눔중을 만들고 경시적으로 병변의 조직학적 소견의 변화와 효소면역법에 의한 혈청 항체가를 관찰하여 이 질병의 병소 형성 과정을 관찰하였다. 뱀에서 얻은 충체를 5마리씩 마우스에 경우 감염시키고, 1,24 및 10주와 6개월 후에 도살하여, 충체의 분포와 수를 육안으로 확인하고 피하 감염부위를 조직표본으로 만들어 현미경으로 관찰하였다. 이와 동시에 효소면역 법을 이용하여 혈청 내 항스파르가눔 IgG 항체가를 측정하여 아래의 결과를 얻었다. 1. 충체 회수울은 감염 후 1주와 2주에 각각 80% 이었으나 조금씩 증가하여 6개월에는 100%이고, 전체 평균 88.3%가 회수되었다. 충체의 대부분은 목파 몸통의 피하조직에서 검출되었고 피하조직에서 골격근에까지 침범 한 것도 일부 관찰되었다. 감염 기간이 경과할수록 충체가 성장하여 고 무게가 처음에 평균 7.3 mg에서 6개월 후에는 47.1 mg으로 증가하였다. 감염기간과 충체의 분포와는 차이가 없었다. 2. 감염 후 1주와 2주에 충체 주위에 염증세포가 국한되어 소수 관찰되었으나, 충체가 지라간 부위로 추측되 는 곳에 많은 세포가 관찰되기도 하였다. 감염 4주 이후에는 충체가 있는 조직 내의 공간 주위로 파괴된 조직세 포와 중성구, 호산구, 림프구등의 염증세포가 많이 침윤되어 있고, 일부에서 삼출액도 관찰되었다. 감염 후 4주 와 10주에는 충체와 이에 인접한 염중세포층의 주위를 조직구(histiocytes)와 섬유다세포가 충을 이루어 둘러싸고 있었다. 감염 후 6개월에 관찰한 바, 대부분의 충체 주위에서 염증세포가 줄고 섬유소에 의한 충낭(worm capsule)이 얇게 형성되어 있었다. 그러나 이 때에도 충체 주위에 농양을 만들거나 심한 염증소견을 보인 부분도 있었다. 모든 감염 군에서 충체는 형태학적으로 온전하고 칼슘 과립을 갖고 있었다. 3. 혈칭 내 IgG 항체가는 평균 흡광도가 감염 후 1주에 0,108, 2주에 0.141이었으나 4주에 0.456, 10주에 0.955, 6개원에 1.060으로 증가하였다. 이상의 결과를 종합하면 스파르가눔 감염 추 4주가 경과하면 염증반응이 강하게 나타나고, 조직구와 섬유세 포가 관찰되기 시작하며, 혈청 내 IgC 항체가도 유의하게 증가함을 알 수 있었다. 충체가 이동하지 않을 경우에 약 10주가 경과하면 충체 주위에 섬유소가 증식하여 충낭을 형성하고 이 기간 이후에는 항체가의 증가도 완만 하였다.

• 농촌의학ㆍ지역보건
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• 제12권1호
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• pp.117-123
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• 1987

Detection of IgG antibody in clonorchiasis has been accomplished through various serodiagnostic procedure including complement fixation test, gel diffusion test, indirect fluorescent antibody test, indirect hemagglutination test etc. In this report enzyme immunoassay (ELISA) and indirect hemagglutination test (IHA) were used to determine IgG serum antibody levels before and after therapy with praziquantel. Briefly, sera from 62 cases of confirmed human clonorchiasis were examined before and after treatment with praziquantel. Among 62 cases treated 25 cases were categorized as completely cured groups by formalin-ether and careful examination of 4 cellophane thick smered slides at 18 months after treatment. The sera of 25 cases of cured groups were examined again by ELISA and IHA, and com-pared to the previous data. The results obtained were as follows; 1) Sensitivity of IHA test was 83.6% when cut-off titer of 1:8 was applied. No sera obtained from 10 normal healthy control showed positive reaction. 2) Twenty cases (80.0%) out of 25 cured one showed negative results by IHA at 18 months after treatment. 3) Although 5 cases showed positive titer even 18 months after treatment 3 cases of them showed decreased antibody titer. However 2 cases did not show any response. 4) Even though almost all cases showed de- creased ELISA value, only 11 cases (44.0%) out of 25 patients showed negative results by ELISA at 18 months after treatment. In conclusion, it is suggested that, while IgG ELISA for detecting long persisting antibody was more sensitive than IHA, IHA results more conclusively indicated effective treatment in clonorchiasis by negative conversion than did the results of ELISA.