• Title/Summary/Keyword: Antibody test

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Development of a Kit for Diagnosing AtCYP78A7 Protein in Abiotic-tolerant Transgenic Rice Overexpressing AtCYP78A7 (AtCYP78A7 과발현 환경스트레스 내성 형질전환 벼의 단백질 진단 키트 개발)

  • Nam, Kyong-Hee;Park, Jung-Ho;Pack, In-Soon;Kim, Ho Bang;Kim, Chang-Gi
    • Journal of Life Science
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    • v.28 no.7
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    • pp.835-840
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    • 2018
  • Quantitative determination of the protein expression levels is one of the most important parts in assessment of the safety of foods derived from genetically modified (GM) crops. Overexpression of AtCYP78A7, a gene encoding cytochrome P450 protein, has been reported to improve tolerance to abiotic stress, such as drought and salt stress, in transgenic rice (Oryza sativa L.). In the present study, an enzyme-linked immunosorbent assay (ELISA) kit for diagnosing AtCYP78A7 protein including AtCYP78A7-specific monoclonal antibody was developed. GST-AtCYP78A7 recombinant protein was induced and purified by affinity column. Four monoclonal antibodies (mAb 6A7, mAb 4C2, mAb 11H6, and mAb 7E8) against recombinant protein were also produced and biotinylated with avidin-HRP. After pairing test using GST-AtCYP78A7 protein and lysate of rice samples, mAb 4C2 and mAb 7E8 were selected as a capture antibody and a detecting antibody, respectively, for ELISA kit. Product test using rice samples indicated that percentages of detected protein in total protein were greater than 0.1% in AtCYP78A7-overexpressing transgenic rice (Line 10B-5 and 18A-4), whereas those in negative control non-transgenic rice (Ilpum and Hwayoung) were less than 0.1%. The ELISA kit developed in this study can be useful for the rapid detection and safety assessment of transgenic rice overexpressing AtCYP78A7.

Radioimmunoassay for Determination of Serum Macrophage Migration Inhibitory Factor (혈중 대식세포 유주 저지 인자 측정을 위한 방사면역측정법)

  • Lee, Tae-Sup;Shin, Seok-Hwan;Song, Jee-In;Woo, Kwang-Sun;Chung, Wee-Sup;Choi, Chang-Woon;Lim, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.6
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    • pp.532-539
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    • 2004
  • Purpose: There has been a renewal of interest in Macrophage migration inhibitory factor (MIF), especially correlation in pathogenesis of sepsis by many infectious diseases and in regulation of host inflammatory and immune response. We developed immunoradiometric assay (IRMA) to determine serum human MIF concentration. Materials and Methods: The IRMA system utilizes solid phase bound monoclonal anti-recombinant human MIF (rhMIF) antibody as a capture antibody, biotinylated polyclonal anti-rhMIF antibody as a detector antibody. We applied with rhMIF that concentration of standard solutions increased from 0 ng/ml to 100 ng/ml. We used $^{125}I$-streptavidin (SA) as radiotracer to determination of rhMIF concentration. Streptavidin was labeled with $^{125}I$ by Chloramine-T method and $^{125}I$-SA was purified by ultracentrifugation. $^{125}I$-SA stability was evaluated by ITLC analysis at $4^{\circ}C$ and room temperatures until 60days. To validate IRMA system for MIF, we experimented intra-assay and inter-assay coefficients of variation, recovery test and dilution test. Results: Radiolabeling yield of $^{125}I$-SA was 87% and purified $^{125}I$-SA retained above 99% radiochemical purity. $^{125}I$-SA showed above 93% stability in $4^{\circ}C$ until 60days that it is good for immunoradiometric assay as radiotracer. Plotted standard dose response curve showed that increased concentration of rhMIF linearly correlated (R2=0.99) with bound radioactivity of $^{125}I$-SA. The highest intra- and inter-assay coefficients of variation were 5.5% and 7.6%, respectively. The average of recovery of MIF in samples was 102%. In dilution test, linear response curves were obtained (R2=0.97). Conclusion: Radioimmunoassay using $^{125}I$-SA as radiotracer thought to be useful for the determination of serum MIF concentration, and further, its data will be used to evaluate the correlation between clinical significance and serum MIF concentration in patients with various inflammatory diseases.

Prospective Observation Study on Hyperthyroidism Patients Treated with Korean Medicine (한의치료를 받은 갑상선 기능 항진증 환자에 대한 전향적 관찰연구)

  • Choi, Yu-jin;Shin, Seon-mi;Han, Yang-hee;Ahn, Se-young;Cho, Chung-sik
    • The Journal of Internal Korean Medicine
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    • v.39 no.1
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    • pp.84-96
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    • 2018
  • Objective: This is aprospective clinical case study that includes the administration of Ahnjeonbaekho-tang (AJBHT) to patients who have suffered from Graves' disease for the 8 weeks of the clinical research. Methods: Without administration of an antithyroid drug, AJBHT was administered 3 times a day for 8 weeks. In Case 1, AJBHT without modification was administered and in Case 2, in accordance with the patients' symptoms, a dose of Gypsum Fibrosum was increased by 4g from the beginning of week 2, and further increased by 16 g, for a total of 20 g between week 6 and week 8. Body mass index (BMI), thyroid function test (TFT), thyroid autoantibody test, complete blood cell count (CBC), and blood chemistry (BC) tests were conducted at an interval of 4 weeks, for a total of 3 times during the study. The clinical manifestations, a spectrum of symptoms of the patients, was observed by Wayne's Index, Euro-QOL-5 Dimension (EQ-5D), and Perceived Stress Scale (PSS). Results: In both cases, an acceptable significance of reduction of value in T3, fT4, Thyroid stimulating antibody (TSAb) and Thyroid stimulating hormone receptor antibody (TSH-R-Ab) was observed. After treatment, in TFT, T3 decreased by 31.66%, fT4 decreased by 32.82% in Case 1, and T3 decreased by 43.42%, and fT4 decreased by 37.32% in Case 2. In the thyroid autoantibody test, TSAb decreased by 7.59%, and TSH-R-Ab decreased by 53.19% in Case 1, and TSAb decreased by 33.45%, TSH-R-Ab decreased by at least 7.75% in Case 2. Besides this, there was a decreasing trend of Wayne's index, and a loss of typical symptoms of hyperthyroidism also declared the efficacy. Conclusion: From these results, AJBHT is very effective in the regulation of TFT and improving the symptoms of hyperthyroidism, and is also expected to be an effective alternative to antithyroid drugs for patients who have side effects or drug intolerance.

Significance of Toxoplasma Antibody Titers by Indirect Latex Agglutination Tests in Pregnant Women and Pelvic Tumor Patients (임산부 및 골반장기 종양환자에서의 Toxoplasma Latex 응집항체치의 의의)

  • Choe, Won-Yeong;Choe, Hyeong-Rak;Na, Jong-Gu
    • Parasites, Hosts and Diseases
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    • v.23 no.2
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    • pp.300-304
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    • 1985
  • A total of 377 pregnant women, 43 pelvic tumor patients and 80 of multiphysic health center persons as controls were examined by indirect latex agglutination test in order to evaluate Toxoplasma antibody titers at Kang-Nam St. Mary's Hospital in Seoul. Throughout this survey, 1 : 32 or more titers of diluted sera were regarded as positive. 1. The 377 samples of test sera in pregnant women showed negatives in 319 cases (84.6%), 1 : 2 in 44 cases (11.7%), 1:4 in 9 cases (2.4%), 1:8 in 2 cases (0.5%), 1:16 in 1 case (0.3%) and 1 : 32 in 2 cases (0.5%) respectively. 2. The 43 samples of test sera in pelvic tumor patients showed negatives in 29 cases (67.4%), 1:2 in 8 cases(18.6%), 1:4 in 1 case(2.3%), 1:16 in 2 cases (4.7%), 1:32 in 1 case(2.3%) and 1 : 128 in 2 cases (4.7%). 3. The 80 samples of test sera in multiphysic health center persons as controls negatives in 56 cases (70.0%), 1.2 in 19 cases (23.8%), 1:4 in 3 cases (3.8%), 1:8 in 1 case (1.3%) and 1 : 128 in 1 case (1.3%). 4. Among total 420 study cases, 5 cases (1.2%) showed Positives, and they were 2 cases (0.5%) of pregnant women and 3 cases (7.0%) of pelvic tumor patients. 5. One case (1.3%) out of 80 control sera showed positive result.

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Seroepidemiology of helminthic diseases in Yanbian of the People's Republic of China (중국 연변지역 주요 기생충질환에 대한 혈청역학적 조사)

  • Joo, Kyoung-Hwan;Quan, Fu Shi;Kwon, Hye-Soo;Cho, Sung-Weon;Li, Shun Yu
    • Journal of agricultural medicine and community health
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    • v.23 no.2
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    • pp.275-285
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    • 1998
  • Antibody responses(IgG) to Paragonimus westermani. Clonorchis sinensis, Cysticercus cellulosae, Sparganum Anisakis simplex, Toxocara canis and Trichinella spiralis were studied. The ELISA technique was performed to determine the prevalence of above helminthic diseases. 975 cases obtained from Yanbian of China during October, 1995 to July, 1997 were examined with a positive antibody titer of 5.74% in clonorchiasis, 4.92% in paragonimiasis, 1.54% in cysticercosis. 8.51% in sparganosis, 1.85% in anisakiosis, 12.51% in toxocariasis, and 7.08% in trichinosis respectively. And 23.87% in showed positive antibody titer at least one of the seven helminths. The differences of the age and sex in the positive sera were analysed by the Chi-squared test and the level of significance accepted was p<0.05. The significant differences in positive antibody production were P.W.(p<0.01). C.S.(p<0.01), A.S.(p<0.05). T.C.(p<0.001), and T.S.(p<0.01) respectively in age groups. sparganosis(p<0.05) in sex groups. Other parasites showed that there were no significant differences among age groups and sex groups(males and females). Higher positive antibody rate of C.S. and P.W. occured in the 50-59 years old and those of T.C. and T.S. happened in the 20-29 years old. Patients of internal disease showed more positive antibody titer, that is to say, there was significant difference between positive rate of internal diseases and that of control (p<0.01. p<0.05) in 6 helminths except cysticercosis. The result showed that some cross reactions existed among nematodes, and the developed techniques(EITB) should be done for a correct diagnosis. Also the prevalence of some important helminths may be evaluated from the result and it would be a basic data for controlling parasitic diseases in Yanbian.

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A Study on the Antigen Characteristics of Rhodotorula rubra (Rhodotorula rubra의 항원특성에 관한 연구)

  • Kwon, Hyuk-Ku;Lee, Jang-Hoon;Ryeon, Kon
    • Journal of Environmental Health Sciences
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    • v.28 no.5
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    • pp.28-34
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    • 2002
  • Antigenicity of Rhodotrula rubra isolated from pulmonary tissue of pulmonary tuberculosis patients was studied by means of agglutination reaction with R. rubra whole cell antiserum. And the serological reactivity of crude polyfac charide from R. frubra, Candida albicans, Candida tropicalis, Candida, glabrata, and Saccharomyces cerevisiae ATCC 26603 with antiserum to R. rubra whole cell was studied by means of immunodiffusion test. R. rubra showed stationary phase after 48h when it was cultured in GYEP broth. While agglutinogen titer was 1:64 at lag phase, agglutinogen titer was 1 :256 after 20h. After growth of R. rubra on different 11 media, nutritional environment showed similar agglu-tination reartivity. The agglutinogen titer of C. albicans, C. tropicalis, C. giabrata, which were isolated from patient's expectoration, to R. rubra antiserum by means of agglutination reaction were 1:16, respectively. But, Sacch. cervisiae ATCC26603 was negative. Those results were lower than that of R. rubra agglutinogen titer 1:256. As a result of immu-nodiffusion test with crude polysaccharide extracted from cell wall of R. rubra, C. albicans, C. tropicalis, C. glabrata, Sacch. cervisiae ATCC26603, precipitin line was found only with R. rubra, of which antibody titer was 8.

Studies on the production and characterization of monoclonal antibodies against bovine rotaviruses isolated in Korea (소 로타바이러스(국내분리주)에 대한 단크론항체 생산 및 특성에 관한 연구)

  • Ahn, Jae-moon;Cho, Sun-hee;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.36 no.2
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    • pp.395-403
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    • 1996
  • Monoclonal antibodies(MAbs) against field isolates of the bovine rotavirus A strain(G6), V strain(G10) and reference I-801 strain(G8) were produced and characterized. Six MAbs(4C2, 4D9, 5E1, 5E7, 5D5, 3E4) against A strain had neutralizing activity and reacted only with the G6 bovine rotaviruses determined by fluorescence focus neutralization (FFN) test. Otherwise, five neutralizing MAbs(1G2, 2G6, 5E2, 5E12, 5H7) against I-801 strain neutralized the G6 and G8 bovine rotaviruses. Five non-neutralizing MAbs(5F12, 7F12, 5E11, 2A11, 2B12) were VP6-specific and cross-reacted with all bovine and porcine rotaviruses examined by fluorescence antibody(FA) test. None of the MAbs reacted with bovie viral diarrhea virus(BVDV) and bovine coronavirus(BCV) determined by FA and FFN test.

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Enzyme-linked immunosorbent assay for detection of bovine antibody to Brucella abortus (축우 부루셀라병의 ELISA 진단법에 관한 연구)

  • Lim, Yoon-kyu;Lee, Doo-sick;Park, Jun-hong;Yang, Ki-chun;Kim, Seung-ho;Kim, Kong-sick;Hyun, Kwan-jong;Kim, Woo-tack;Lee, Yong-soon
    • Korean Journal of Veterinary Research
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    • v.33 no.1
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    • pp.131-135
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    • 1993
  • Enzyme-linked Immuno sorbent Assay (ELISA) for the serological diagnosis of Brucella abortus was developed and compared with plate agglutination test. Cell wall antigen was extracted from Brucella abortus 1119-3 by sonication and with a sodium deoxychlate solution. Optimum protein concentration of coating antigen were $0.4{\mu}g/100{\mu}{\ell}$ protein on each microtiter plate well. Horse radish peroxidase (HRP) labeled protein-G was used as a tracer of reacted antibodies. ELISA confirmed the agreeable results of 40 cases out of 43 cases by plate aggulutination test. ELISA diagnosed positive cases(10 out of 12) and negative cases (1 out of 12) with dubious sera by plate agglutination test. From this results ELISA could be used for the early diagnostic tools of Brucellosis in cattle.

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Pulmonary Artery Aneurysm in Behcet`s Disease - Report of a Case - (Bechet씨 병에 발생한 폐동맥류: 치험 1례)

  • 김선한
    • Journal of Chest Surgery
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    • v.25 no.4
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    • pp.412-417
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    • 1992
  • Pulmonary artery aneurysm is Behcet`s disease is rare and can be fatal due to rupture. We experienced a case of pulmonary artery aneurysm in Behcet`s disease. The patient was 21 year old woman who was adimitted with three month history of dyspnea, fever and cough. On examination, she had aphthous ulcer in the mouth and erythema nodosum on the left popletial fossa and forearm, but didn`t have any lesion at eyes and genitalia. The latex fixation test for rheumatoid factor, VDRL test for syphillis, antinuclear antibody and LE cell test were all negatives. The third and fourth components of complement in the serum, serum immunoglobulin concentrations[IgG, IgM, IgA] were within normal range. The chest radiography revealed a 5x6cm sized radiopaque mass density in the left hilar region. Two months later, the mass was enlarged to 6x7cm. The IV-DSA showed a single aneurysm at the proximal part of left lower lobe artery with lingular segment artery and no distal perfusion by thombotic obstruction. The steroid therapy was done for a month, but symptoms not improved. We performed resection of lingular segment and lower lobe including the aneurysm. The microscopic findings of the operative specimen were intimal hyperplasia and fragmentation of the internal elastic fibers. She was improved without remarkable event, except infection of the operative wound.

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Experimental study of antigenicity test of Sweet Bee Venom in Guinea Pigs (기니픽을 이용한 Sweet Bee Venom의 항원성 평가)

  • Cho, Byung-Jun;Kwon, Ki-Rok
    • Journal of Pharmacopuncture
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    • v.14 no.4
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    • pp.23-32
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    • 2011
  • Objectives: This study was performed to examine the antigenic potential of pure melittin (Sweet Bee Venom - SBV) extracted from the bee venom by utilizing protein isolation method of gel filtration. Methods: All experiments were conducted at Biotoxtech (Chungwon, Korea), authorized a non-clinical studies institution, under the regulations of Good Laboratory Practice (GLP). Antigenic potential of SBV was examined by active systemic anaphylaxis (ASA) and passive cutaneous anaphylaxis (PCA) in guinea pigs. SBV was subcutaneously administered at 0.07 and 0.28mg/kg and also as a suspension with adjuvant (Freund's complete adjuvant: FCA). Ovalbumin (OVA) as a suspension with adjuvant was used to induce positive control response ($5mg/m{\ell}$-FCA). Results: 1. In the ASA test, experimental groups showed some symptoms of anaphylaxis like piloerection, hyperpnea and staggering gait. 2. In the PCA test, low dosage group did not show any antibody responses, whereas high dosage group showed positive responses. 3. In the weight measurement and clinical observation, experimental groups didn't show any significant changes compared with control group. 4. In the autopsy of body, the abnormalities of lung were detected in the corpse. This means that the cause of death may induced anaphylactic shock. Conclusions: Above findings suggested that SBV had antigenic potential in guinea pig. Further studies on the subject should be conducted to yield more concrete evidences.