• Title/Summary/Keyword: serum IgG antibody

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Use of the enzyme-linked immunosorbent assay for the detection of toxoplasmosis in swine (ELISA를 이용한 돼지 톡소플라스마병의 조기 진단에 관한 연구)

  • Suh, Myung-deuk;Jang, Dong-hwa;Joo, Hoo-don
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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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.

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Effects of Deerhorn Extracts on the Antibody Productions against Escherichia coli Antigen (녹용 추출액이 대장균 항체생산에 미치는 영향)

  • 이형환;제갈승주
    • Korean Journal of Microbiology
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    • v.21 no.1
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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.

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Detection of Antibody to Infectious Laryngotracheitis Virus by Enzyme Linked Immunosorbent Assay (효소면역법에 의한 닭 전염성 후두기관염 바이러스 항체 측정에 관한 연구)

  • 임숙경;위성하;최정옥;고홍남
    • Korean Journal of Veterinary Service
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    • v.15 no.1
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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.

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High Levels of Antibodies to Plasmodium falciparum Liver Stage Antigen-1 in Naturally Infected Individuals in Myanmar

  • Lee, Hyeong-Woo;Moon, Sung-Ung;Kim, Yeon-Joo;Cho, Shin-Hyeong;Lin, Khin;Na, Byoung-Kuk;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.46 no.3
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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.

Effects of Pinelliae Rizoma (PR) on asthma induced intra-nasal instillation of ovalbumin in mice (반하가 난황으로유발된 생쥐의 알레르기성 천식에 미치는 영향)

  • Jeong, Dong-Hwan;Kim, Jong-Han;Park, Su-Yeon;Choi, Jeong-Hwa
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.21 no.1
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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.

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Characterization of a Peptide Antibody Specific to the Adenylyl Cyclase-Associated Protein of Acanthamoeba castellanii

  • Kim, Min-Jeong;Lee, Hae-Ahm;Quan, Fu-Shi;Kong, Hyun-Hee;Moon, Eun-Kyung
    • Parasites, Hosts and Diseases
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    • v.60 no.1
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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.

Analysis of Human Serum Amyloid A-1 Concentrations Using a Lateral Flow Immunoassay with CdSe/ZnS Quantum Dots (Human Serum Amyloid A-1 단백질 농도 분석을 위한 CdSe/ZnS 양자점 기반의 Lateral Flow Immunoassay 방법 개발)

  • Fajri, Aidil;Goh, Eunseo;Lee, Sanghyuk;Lee, Hye Jin
    • Applied Chemistry for Engineering
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    • v.30 no.4
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    • pp.429-434
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    • 2019
  • A lateral flow immunoassay platform utilizing antibody functionalized water soluble CdSe/ZnS semiconductor quantum dots (QDs) was developed for the analysis of human serum amyloid A-1 (hSAA1) in a buffer solution. hSAA1 was chosen as a target protein because it is regarded as a potential biomarker associated with early diagnosis and prognosis in patients of lung cancer. The immunoassay strip on a nitrocellulose membrane was fabricated by spraying two lines composed of a test line with a monoclonal antibody against hSAA1 (10G1) (anti hSAA1) and a control line of anti-chicken IgY. While the CdSe/ZnS QDs synthesized in an organic phase were transferred to a water phase by ligand exchange using carboxylic acid modified alkane thiol. The QDs was then conjugated to monoclonal antibody against hSAA1 (14F8) [anti hSAA1 (14F8)] and used as a fluorescent detection probe. The sequential lateral flow of hSAA1 in different concentration and QDs-anti hSAA1 (14F8) complex allowed to form the surface sandwich complex of anti hSAA1 (10G1)/hSAA1/QD-anti hSAA1 (14F8), which was then analyzed using fluorescence microscope. A 100 nM concentration of hSAA1 protein can be detected by naked eyes under an optimized lateral flow buffer condition with a sensing time of 5 mins.

Immunoradiometric Assay using Monoclonal Antibody Against Human Serum Transferrin Receptor for Diagnosis of Iron Deficiency (사람 혈청 트란스페린수용체의 단클론 항체를 이용한 방사면역측정과 철영양상태의 진단)

  • 김승렬
    • Journal of Nutrition and Health
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    • v.29 no.9
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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.

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The changes of histopathology and serum anti-sparganum IgG in experimental sparganosis of mice (마우스 피하 스파르가눔증에 있어서 감염 경과에 따른 조직병리학적 병변 및 혈청 항체가의 변화 양상)

  • 홍성태;김계정
    • Parasites, Hosts and Diseases
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    • v.27 no.4
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    • pp.261-270
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    • 1989
  • The present study is intended to observe the chronologic changes of experimental sparganosis by histopathological observation and detection of circulating anti-sparganum IgG antibody using ELISA. Each of 25 mice was infected with aye spargana, and they were examined after 1, 2, 4, 10 weeks or 6 months from infection. The followings are summarized results. - 1. The plerocercoids were detected in the subcutaneous tissue of the trunk, neck or axilla, but a few often extended into the skeletal muscle. The recovery rates were 72% at the first week, 80% at the second week, 95% at the fourth week, 92% at the tenth week and 100% at .the sixth month. The larvae grew slowly in both length and weight until 6 months. 2. Histopathologically, most of the larvae were observed alive in the soft tissue or skeletal muscle. Numerous eosinophils, neutrophils, Iymphocytes and plasma cells were infiltrated focally around the worms by the second week, but they surrounded the worms to form a layer of inflammatory reaction after 4 weeks of infection. Also histiocytes and fibroblasts began to appear around the inflammatory cells at 4 weeks. After 10 weeks, the worms encircled by a thin fibrous layer were found. After 6 months, the worms were surrounded by either fibrous tissue or active inflammatory cells. The inflammation looked more severe in the tracks left by the worms, rather than around the worms. 3, The level of anti-sparganum IgG antibody in the serum showed an increase by the fourth week, and a rapid and continuous increase was observed thereafter by the tenth week after infection. The high level of the IgG antibody was maintained up to 6 months forming a plateau curve. The present results suggest that the tissue reaction and antibody production in subcutaneous sparganosis become distinctive by the fourth week after infection.

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Changes of Serum antibody Levels by Indirect Hemagglutination Test after Treatment in Clonorchiasis (IHA(Indirect hemagglutination test)를 이용한 간흡충증(肝吸虫症)의 면역혈청학적(免疫血淸學的) 진단(診斷)에 관한 연구(硏究) -ELISA법과의 비교(比較) 및 치료(治療) 전후(前後) 의 혈청항체가(血淸抗體價) 변동(變動)에 관하여-)

  • Joo, Kyoung-Hwan;Kim, Jong-Seong;Chung, Myung-Sook;Rim, Han-Jong
    • Journal of agricultural medicine and community health
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    • v.12 no.1
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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.

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