• Title/Summary/Keyword: indirect-ELISA

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Detection of PCB77 by Indirect Competitive Enzyme-linked Immunosorbent Assay in Sea Sediment Samples

  • Chen, Han-Yu;Zhuang, Hui-Sheng;Yang, Guang-Xin
    • Bulletin of the Korean Chemical Society
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    • v.34 no.3
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    • pp.922-926
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    • 2013
  • 3,3',4,4'-Tetrachlorobiphenyl (IUPAC PCB77) is one of seven indicative polychlorinated biphenyls (PCBs) in the surface sediments. The current study presents a novel polyclonal antibody for the determination of the PCB77 using indirect competitive enzyme-linked immunosorbent assay. Under optimum conditions, PCB77 was determined within the concentration range of 0.01-100 ${\mu}g\;L^{-1}$, with a detection limit of 0.057 ${\mu}g\;L^{-1}$. The assays were tested for their cross-reactivity profiles using 3 selected congeners and 4 Aroclor products. The assays were highly specific for coplanar PCB congeners, but less specific for Aroclor1248. The spiked recoveries from five sediment samples were 86%-114% for PCB77 from ELISA, which were satisfactory. The current study demonstrated that the developed antiserum and immunoassay procedure can be used to detect PCB77 in environmental samples. The results of the sediment analysis were confirmed by conventional GC/ECD.

Detection of antibodies against infectious Borna disease virus -a comparison of three serological methods- (보르나병 바이러스 항체검출을 위한 연구 -세 가지 혈청진단법의 비교-)

  • Lee, Du-sik
    • Korean Journal of Veterinary Research
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    • v.32 no.1
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    • pp.57-61
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    • 1992
  • To determin the accuracy of serological methods in detecting Borna-disease(BD) viral antibodies, 273 experimentally infected rabbit sera were compared by using indirect immunofluorescence antibody test(IFA), serum neutralization test(SN) and enzyme-linked immunosorbent assay(ELISA). One hundred twenty-three serum samples had BD viral antibodies detected by IFA. CELISA antibodies to BD virus were also present in the same one hundred twenty-three serum samples. However, neutralization test antibodies to BD virus were present in 27 of the in rabbit serum samples. Neutralization test was sensitive in comparison with KFA and CELISA. In comparison with IFA, CELISA was both sensitive and specific in detecting BD viral antibodies. These results extend observations made with laboratory animals to the diagnosis of naturally infected animals.

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Origin of Milled Wood Lignin : An Immunocytochemical Approaches in Combination with Transmission Electron Microscopy (면역전현법(免疫電顯法)에 의한 마쇄(磨碎) 리그닌의 기원(起源)에 관한 연구(硏究))

  • Kim, Yoon-Soo;Koh, Hong-Bum
    • Journal of the Korean Wood Science and Technology
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    • v.24 no.1
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    • pp.68-74
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    • 1996
  • Present work was undertaken to investigate the origin of milled wood lignin(MWL) in the wood cell wall using immunocytochemical techniques, which can provide the information on the localization of specific antigens(MWL in the present study) to be examined. Spruce MWL dissolved in DMSO and emulsified with Freund adjuvant was injected directly into the mouse spleen. The animals were boostered at two-week intervals after the initial immunization. Blood samples were purified in standard procedures. The characteristics of antibodies against MWL were tested by indirect ELISA. Visualization of MWL was carried out using conventional indirect immunogold-labelling methods on the ultrathin sections of spruce wood. Immuno-TEM observations showed that the immunogold probes were selectively attached to secondary cell walls of spruce wood. The most intense labelling was frequently observed in the S2 layer. In contrast, gold labelling in the lignin-rich regions, such as middle lamella and cell corner was not found. The immuno-TEM provides an indication that spruce MWL originates from the S2 layer.

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Indirect enzyme linked immunosorbent assay for the diagnosis of brucellosis in cattle

  • Rahman, Siddiqur;Huque, Fazlul;Ahasan, Shamim;Song, Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.33 no.2
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    • pp.113-119
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    • 2010
  • Brucellosis is a major zoonosis caused by Gram negative facultative intracellular bacterial organisms of the genus Brucella that are pathogenic for a wide variety of animals and human beings. Because of its economic impact on animal health and the risk to the human population,most countries have a brucellosis control program. Brucellosis is also an economically important andprevalent disease in Bangladesh. The accurate and prompt diagnosis is very important in controlling and eradicating of the disease in animals. The present study was undertaken to determine the seroprevalence of brucellosis in cattle in Mymensingh and Patuakhali district of Bangladesh. A total of 120 serum samples were collected from the two districts along with a questionnaire related to the epidemiology of the disease. The sampleswere screened by using slow agglutination test and conformed by indirect enzyme linked immunosorbent assay. The overall seroprevalence of brucellosis in cattle was 5% and it was observed that, a higher prevalence of Brucella was found in female than male, through natural breeding than artificial insemination (AI) and animal above 4 years old are highly susceptible than younger ones. Higher prevalence was found in aborted animals in comparison with non aborted animal. Finally, the study revealed that the female animal has more susceptible to brucellosis and healthy semen should be used for AI.

Evaluation of the antibodies against Pseudomonas tolaasii and Pseudomonas agarici for Immunoassays and Its Application (항 Pseudomonas tolaasii 및 Pseudomonas agarici 항체의 면역측정법을 위한 평가와 그 이용)

  • Jeon, Nak-Beom;Lee, Hyang-Burm;Shon, Dong-Hwa;Yu, Seung Hun
    • Research in Plant Disease
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    • v.9 no.2
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    • pp.94-98
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    • 2003
  • To produce antibodies against Pseudomonas tolaasii and P. agarici, lyophilized P. tolaasii and P. agarici ($5{\times}10^7$ cfu/ml) and Freund, s adjuvant were immunized into rabbits 4 times. The specificity and sensitivity of the antibodies were evaluated by immunodiffusion test and indirect enzyme-linked immunosorbent assay (id ELISA). The ${\alpha}$-P. tolaasii antibody was very specific only against P. tolaasii, while ${\alpha}$-P. agarici antibody was not specific and showed a high cross reactivity toward P. tolaasii with detection limit concentration of $2{\times}10^3$ cfu/ml. However, the cross reactivities of ${\alpha}$-P. agarici antibody toward the related species including P. reactans were very low. Our results showed that ${\alpha}$-P. tolaasii and ${\alpha}$-P. agarici antibodies against P. tolaasii and P. agarici, respectively, might be useful for rapid and simple detection of the causal agents of bacterial brown and yellow blotches in cultivated oyster mushrooms.

A toxoplasmic uveitis case of a 60-year-old male in Korea

  • Kim, Min-Ho;Choi, Yeun-Kyoung;Park, Yun-Kyu;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.38 no.1
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    • pp.29-31
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    • 2000
  • A toxoplasmic uveitis case was reported on the focus of impairment of pathological findings and serological antibody titers after chemotherapy. A chief complaint of a 60-year-old male was a decreased and blurred vision in his right eye for 2 weeks after experiencing tremendous stress and fatigue. A steroid therapy for 3 weeks was not effective and the retinal lesion became necrotic. Anti-Toxoplasma gondii antibody titer was checked to be a strong positive by both ELISA and indirect latex agglutination assay (lLA). He was treated with Fansidar F for 8 weeks. His vision improved as the necrotic lesion healed by scarring, but the antibody titers still remained very high without any signs of negative conversion. It is suggested to be a recurrent case of the past asymptomatic infection by presumed immune suppression caused by excessive stress.

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Effects of Polyphosphates on the Growth and T-2 Toxin Production of Fusarium sporotrichioides M-1-1 (인산염이 Fusarium sporotrichioides M-1-1 성장과 T-2 toxin 생성에 미치는 영향)

  • 장덕화;송재영;김일환
    • Journal of Food Hygiene and Safety
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    • v.10 no.4
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    • pp.199-204
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    • 1995
  • The antifungal effects of polyposphates on the growth and T-2 toxin production of Fusarium sporotrichioides M-1-1 were investigated. The growth of the strain was significantly inhibited in the potatoes dextrose agar medium treated with 1.5% polyphosphates or more. When we checked T-2 toxin by the indirect competitive ELISA, the strain produced 11.25 ug/ml and 10.90 ug/ml levels of T-2 toxin rice and corn containing 50% moisture contents, respectively. However, T-2 toxin was little detected in rice medium and corn medium with 1.5% polyphosphates addition for short(14 days) and prolonged incubation time(45 days). We also observed the destruction of cell wall and outflow of cell ingredients with 1% polyphosphates treatment to the strain. Therefore, moisture and polyphosphates greatly effected on the growth and T-2 toxin production of the strain.

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Characterization and diagnosis of YMV-K strains infecting Chinese yam(Dioscorea opposita Thunb. cv. Dung-Gun-Ma) in Korea. (oral)

  • D. K. Kang;H. Y. Shin;J. H. Sung;Park, J. H.;M. U. Chang
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.127.2-127
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    • 2003
  • YMV-K strains were purified from D. oppostita Thunb. tv. Dung-Gun-Ma showing mosaic symptom on their leaves. YMV-K strains were filamentous particles of 780nm in length and induced cytoplasmic disorder such as inclusion body formation. Nucleotide and amino acid sequences of 5'-UTR, Pl and CP of YMV-K strains shared 80.8, 64.7 and 98.3% identity respectively to JYMV J1 in the mean value. Purification of YMV-K strains according to JYMV purification method(S. Fuji) was conducted to product antiserum. With antiserum against YMV-K strains, the Various diagnosis methods such as IC-RT-PCR, DIBA, RIPA and indirect-ELISA were used to detect YMV-K strains in Chinese yam plant.

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LEVELS OF TNF-α,-β, IL-1β, TGF-β1 AND THEIR RELATIONSHIP WITH THE PRESENCE OF SPECIFIC BLACK PIGMENTED BACTERIA IN PERIAPICAL AND PULPAL DISEASES (치근단 및 치수병변 환자에서의 TNF-α와 β, IL-1β 및 TGF-β1의 수준과 근관내 특정 black pigmented bacteria와의 연관성에 관한 연구)

  • Byun, Ho-Young;Lim, Sung-Sam;Park, Dong-Sung
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.1-12
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    • 1999
  • Bacterial infection of the pulp results in the development of a periapical lesion with the concomitant resorption of periapical bone. The cytokines are believed to play an important role in this matter. The purpose of this study was to find the relationship among the presence of black pigmented bacteria, the levels of cytokines(TNF-${\alpha}$, -${\beta}$, IL-$1{\beta}$, and TGF-${\beta}1$), and the amount of bone resorption in periapical and pulpal diseases. For the purpose, the patients were grouped into chronic apical pathosis, acute apical pathosis, acute pulpitis, and a healthy control group. Root canal samples were taken from periapical tissue exudates during routine endodontic treatment, and the venous blood was taken from each patients. The samples were processed to measure local and systemic levels of the cytokines using enzyme linked immunosorbent assay(ELISA). Bacterial content of Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella nigrescens were measured by indirect immunofluorescence method and the size of the periapical lesions were measured from the radiographs. The following results were obtained: 1. The levels of bone resorptive cytokines(TNF-${\alpha}$, TNF-${\beta}$, and IL-$1{\beta}$) in exudates from acute and chronic apical pathoses were significantly higher than those from acute pulpitis and the normal pulps(p<0.05). 2. IL-$1{\beta}$ were the highest among the bone resorptive cytokines in apical pathoses. However, no statistical difference between acute and chronic lesions were found(p>0.05). 3. The levels of TGF-${\beta}1$ in exudates from acute pulpitis and chronic apical pathoses were significantly higher than those from acute apical pathoses and the normal pulps(p<0.05). However, there were no significant correlations among the levels of bone resorptive cytokines. 4. The levels of TNF-${\beta}$ in serum were significantly higher than those from the exudates while serum TGF-${\beta}1$ concentrations were significantly lower(p<0.05). 5. Exudates from the canals in which the P. nigrescens were detected showed significantly higher levels of IL-$1{\beta}$ than those from the canals without the microorganism(p<0.05). 6. There were no significant correlations among the levels of the cytokines, the amount of bone destruction, and the presence of acute and chronic symptoms(p>0.05).

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Effect of Reactive Oxygen Species on the Formation of Nε-(carboxymethyl)lysine, an Endproduct of Maillard Reaction of Proteins (단백질의 Maillard 반응의 최종산물인 CML 형성에 미치는 ROS의 영향)

  • Lee, Jong-Sun;Yang, Ryung;Shin, Dong-Bum
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.123-131
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    • 2004
  • The effect of reactive oxygen species (ROS) on the formation of $N^{\varepsilon}$-(carboxymethly)lysine (CML). one of the endproducts in the Maillard reaction of protein (or glycation), was investigated. Glyoxal, a main precursor of CML formation, was produced from both glucose and fructose during their autoxidation. The transition metal ion showed to involve in the formation of glyoxal by the metal catalyzed oxidation, suggesting that ROS accelerated the reducing sugar autoxidation. The stimulative effect of ROS on the autoxidation was more prominent in glucose than in fructose. Polyunsaturated acids (PUFAs) were shown to form glyoxal by peroxidation in proportion to the degree of unsaturation, but ROS did not affect on PUFA peroxidation. Ascorbic acid also lysine (CMHL) in the model system using hippuryl lysine and glucose had a significant effect on ROS, whereas it had no effect on ROS using glyoxal as a reactant. Almost the same trend was obtained by the analysis of antigen coated indirect noncompetitive ELISA using monoclonal antibody (6D12). These data indicated that ROS affected glucose autoxidation as well as mediated both CML and glyoxal formation, but did not affect the reactive compounds such as fructose, PUFAs and ascorbic acid.