• Title/Summary/Keyword: Cross-reactivity

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Diagnosis of bovine cryptosporidiosis by indirect immunofluorescence assay using monoclonal antibody (단세포군항체를 이용한 간접형광항체법에 의한 송아지 작은와포자충증의 진단)

  • Wi, Seong-Hwan;Lee, Jeong-Gil;Ju, Hu-Don
    • Parasites, Hosts and Diseases
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    • v.33 no.2
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    • pp.107-116
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    • 1995
  • Two hybridoma cell lines against Cwptosporinium possum oocysts nFRl-CN911 were produced. The isotype of these 2 monoclonal antibodies (mAbs) was IgG2b (lE7.2) and and IgM (C6). Enzyme immuno-transfer blotting analysis showed that 157.2 reacted specifically to 36 kDa protein and C6 reacted to 67 and 70 kDa proteins. C. pcnlum was bound specifically to the surface region of oocysts by these mobs. No cross-reactivity was observed with tachyzoites of ToxopLosma gonnii and oocysts of Eimeria zuernii,5. bouis and E. canadensis of bovine origin. The indirect immunofluorescence assay (IIF) using mAb C6 was successful with counterstain. With the IIF using mob C6, oocysts appeared as 3 to $5{\mu}m$ spherical objects fluorescing bright apple green against a reddish dark background. The IIF using mAb C6 was agreed in specificity and sensitivity with those of a commercial diagnostic kit. These results demonstrated that the produced mAbs were specific to C. parvum and that the mAb C6 could be used for diagnosis of cryptosporidiosis.

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Accuracy of an Interferon-gamma Release Assay to Detect Active Tuberculosis in Children: A Pilot Study (소아 결핵 진단에서의 인터페론감마 분비 검사의 유용성)

  • Lee, Young Jin;Chun, Peter;We, Ju Hee;Park, Su Eun
    • Pediatric Infection and Vaccine
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    • v.18 no.1
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    • pp.48-53
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    • 2011
  • Purpose : Early diagnosis of active tuberculosis (TB) in children is difficult. The widely used tuberculin skin test has low sensitivity and cross reactivity with non-tuberculous mycobacteria or Bacille Calmette-Gu$\acute{e}$rin vaccination. Interferon gamma release assays have been shown good diagnostic accuracy for active in adults. But studies in children were limited. The purpose of this study was to examine the performance of enzyme-linked immunospot assay (ELISpot) as an initial test in the diagnosis of active tuberculosis in children. Methods : In a hospital-based study, we prospectively examined the performance of ELISPot in 33 children suspected of active TB. TB was confirmed bacteriologically or histologically. Results : Among 33 patients, 9 had active tuberculosis. When tested, they all had a positive test result from the ELISpot. The sensitivity and specificity of the assay were 100% (95% CI, 66.4-100%) and 95.8% (95% CI, 78.9-99.9%) respectively. Conclusion : ELISpot might be an useful and improved clinical diagnostic method for the detection of active TB in children.

Development and Application of Enzyme Immunoassay for Endosulfan Residue Analysis (Endosulfan 과 그 분해산물의 Enzyme Immunoassay에 의한 분석법의 개발과 응용)

  • Suh, Yong-Tack;Shim, Jae-Han;Lee, Kang-Bong
    • Korean Journal of Environmental Agriculture
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    • v.11 no.1
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    • pp.59-66
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    • 1992
  • An enzyme immunoassay(EIA) was developed for the analysis of insecticide endosulfan and its degradation products. The sensitivity and specificity of the antibody produced were examined. Optimal conditions in the ELISA system for residue analysis were also discussed. A mixed suspension of endosulfan-hemocyanin conjugate(ES-KLH) 1.1 mg/ml and Freund's adjuvant was injected subcutaneously to white rabbits and then collected antisera were tested for titers by indirect ELISA(1/24,000). Because of difficulties in the synthesis of endosulfan peroxidase conjugate, amine derivative of endosulfan-diol was synthesized and it showed 40% of conjugate yields(2mg/ml of conjugate). the highest sensitivity obtained enzyme-conjugate was a concentration of 200ng/ml. The detection limit of endosulfan in ELISA system was 5 ppb on the standard curve. In application of ELISA for residue analysis, the recoveries were really 100% both in the spiked soil and apple sample regardless of endosulfan concentration treated. On the other hand, chlorinated hydrocarbons of similar structure with endosulfan showed low cross-reactivity$(2.2%{\sim}29.2%).$

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A Study on the Oxidative Transformation of Quinone Compound using Nanostructured Black-birnessite (나노구조의 블랙-버네사이트를 이용한 퀴논계 화합물의 산화-변환 연구)

  • Harn, Yoon-I;Choi, Chan-Kyu;Shin, Hyun-Sang
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.6
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    • pp.547-554
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    • 2010
  • In this study, new manganese oxide (i.e., black-birnessite) particles with nanostructures were prepared and its physico-chemical properties and oxidative-transformation efficiency on 1,4-naphthoquinine(1,4-NPQ) in the presence of reactive mediator was investigated. The results were also compared with that of the manganese oxide (i.e., brown-birnessite) particles synthesized by classical McKenzie method. Analysis of XRD and SEM data show that the particles are a single phase corresponding to a birnessite-based manganese oxide with cotton ball-like shapes containing nanofibers. In batch experiments, removals of 1,4-NPQ by the black-birnessite follows pseudo-first-order kinetics and the rate constant values obtained are greater about 2.3 times than that of the brown-birnessite in spite of its lower surface area (41.0 vs 19.80 $m^2/g$). The results can be explained by the higher crystallinity and nano structured features of the back-birnessite particles, which give higher reactivity for the removals of the quinone compound. HPLC analysis of the reaction products confirmed that the balck-birnessites removed 1,4-NPQ through cross-coupling reaction in the presence of catechol as a reactive mediator.

The Sensitization Rates for Inhaled Allergens by Skin Prick Test among Some Farmers in Gyeonggi Province, South Korea (피부 단자 검사로 평가한 경기도 일부 농업인의 흡입 알레르겐 감작률)

  • Kim, Hogil;Lee, Ji-Hoon;Roh, Soo-Yong;Lee, HyangSeok;Kwon, Soon-Chan;Lee, Soo-Jin
    • Journal of agricultural medicine and community health
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    • v.40 no.4
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    • pp.240-249
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    • 2015
  • Objectives: Farmers are known to be exposed to a variety of allergens related to the work environment. This study was conducted to determine the sensitization rates as well as South Korea that they are sensitized to certain allergens farmers through the skin prick test. Methods: By targeting a total of 1143 people living in the rural town of Gyeonggi Province, it was conducted a questionnaire containing demographic and occupational risk factors and underwent skin prick tests with 15 types of allergens(including positive and negative controls). Multivariable logistic regression analysis was used to analyze the association between occupational risk factors and skin prick test positivity. Results: Except for the 30 people whose result is invalid, positive rate of the skin prick test was 18.6% in 1,113 people. The species of house dust mite, Dermatophagoides pteronyssinus and Dermatophagoides farinae was the highest at 8.7% and 8.6%. After adjusted by age, gender, smoking and education level, odds ratio of flower plant farmers is 4.467(95% CI: 2.094-9.527) and fruit farmer is 2.275(95% CI: 1.096-4.721). In addition, the rate of sensitization to grass pollen mixture of the flower plant farmers is significantly higher(15.9%, p<0.001) than other allergens. Conclusions: Even farmers, the rate of sensitization to allergens related to the general environment, such as house dust mite is relatively dominant. However, given the presence of potential cross-reactivity between the allergens or distribution showed that the unique aspects of allergen sensitization in the flower growers, occupational cause is not be completely ruled out.

Age-related Changes in Plasma Leptin from Early Growing to Late Finishing Stages of Castrated Holstein Steers: Utilizing Multi-species Leptin RIA

  • Vega, R.A.;Lee, H.G.;Kuwayama, H.;Matsunaga, N.;Hidari, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.5
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    • pp.725-731
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    • 2002
  • This experiment was performed to understand the changes in plasma leptin in association with plasma IGF-1, body weight and ADG from early growing to late finishing stages of Holstein steers. Blood collection was performed by arterial vein puncture at selected monthly ages of 1 (54 kg), 2.6 (103 kg), 7.2 (205 kg), 13.5 (314 kg), 16.9 (414 kg), 22.2 (550 kg), 24.9 (626 kg) and 27.4 months (695 kg). The blood was analyzed for leptin using the multi-species leptin RIA with recombinant bovine leptin (rbleptin) as standard, plasma IGF-1 was also measured using RIA. Against the standard rbleptin, the multi-species Leptin RIA system's sensitivity, cross reactivity, slope and recovery of 41.0 ng/ml rbleptin in plasma were 4.9 ng/ml, 11.22%, -1.396 and 97.8%, respectively. Plasma leptin measured were more than 5.0 ng/ml, which enable multi-species RIA system to investigate plasma leptin in normal growing steers. Body weight resulted to a highly significant second-degree polynomial relationship with plasma leptin (q=0.54, p<0.0001) and plasma IGF-1 (q=0.44, p<0.0001) from 1 to 27.4 monthly ages. However, the second-degree polynomial curve of plasma leptin and IGF-1 differs showing a concave and convex curvilinear relationship, respectively. ADG was not significantly associated to plasma leptin (r=0.06, p>0.05) and plasma IGF=1 (r=0.06, p>0.05) from 1 to 27.4 monthly ages. Low coefficient, but significant associated increase of plasma leptin and IGF-1 (r=0.12, p<0.008) from 1 to 27.4 months was observed. The uncoordinated increases of plasma IGF-1 at growing and plasma leptin at fattening period, may indicate (1) indirect involvement of endogenous IGF-1 on leptin secretion, and (2) IGF-1 level may signify lean and bone accretion while plasma leptin may mirror body fatness across the monthly ages of Holstein steers.

Development of Enzyme-Linked Immunosorbent Assay for the Detection of Fumonisins (효소면역측정법을 이용한 Fumonisin의 검출법 개발)

  • Shon, Dong-Hwa;Hahn, Seong-Min;Lim, Sun-Hee;Lee, Yin-Won;Cho, Sun-Hee;Kang, Shin-Young;Lee, Kyung-Ae
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.119-125
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    • 1996
  • In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishment of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin $B_1(FB_1)$ conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit was tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1:16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigated by the ciELISA, it was very low against $B_3$ (2%) but high against fumonisin $B_2$ (179%). The sensitivity of the ELISAs was also very high, because the detection limit for $FB_1$ was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of $FB_1$ was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of $FB_1$ from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

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Immuno-specificity of Egg Yolk Antibodies against Bovine Rotavirus and Bovine Coronavirus causing Calf Diarrhea (송아지 설사병 주요원인체인 소로타바이러스와 소코로나바이러스에 대한 난황항체 생산 및 면역특이성 분석)

  • Lee, Seong;Woo, Seung-Eun;Lee, Sang-Rae;Kim, Jung-Woo
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.407-412
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    • 2009
  • This study was performed to produce specific egg yolk antibodies against bovine rotavirus (BRV) and bovine coronavirus (BCV) that are major pathogens causing diarrhea in calves. Chickens were immunized with BRV and BCV intramuscularly in the breast muscle by injection 5 times at two weeks interval. At 6 weeks after the first immunization of BRV or BCV, cross reactivity of each serum derived from BRV- or BCV-immunized hens was tested. Each serum antibody against BRV or BCV was reacted with only specific BRV or BCV antigens. Serum and egg yolk-antibody titers of hens against BRV or BCVwere highest at 8~12 weeks after first immunization. Specific serum and egg yolk-antibody titers against BRV were about 104,000 and 107,000, respectively, and those against BCV were about 145,000 and 155,000, respectively. Hemagglutination inhibition titers in the immunized egg yolk antibodies against BRV and BCV were 5,120 and 1,280, respectively, and were ${\geq}8$ times higher than that in non-immunized control. These results suggested that the immunized egg yolk antibodies could effectively neutralize BRV and BCV.

Loop-Mediated Isothermal Amplification Targeting 18S Ribosomal DNA for Rapid Detection of Acanthamoeba

  • Yang, Hye-Won;Lee, Yu-Ran;Inoue, Noboru;Jha, Bijay Kumar;Sylvatrie Danne, Dinzouna-Boutamba;Kim, Hong-Kyun;Lee, Junhun;Goo, Youn-Kyoung;Kong, Hyun-Hee;Chung, Dong-Il;Hong, Yeonchul
    • Parasites, Hosts and Diseases
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    • v.51 no.3
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    • pp.269-277
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    • 2013
  • Amoebic keratitis (AK) caused by Acanthamoeba is one of the most serious corneal infections. AK is frequently misdiagnosed initially as viral, bacterial, or fungal keratitis, thus ensuring treatment delays. Accordingly, the early detection of Acanthamoeba would contribute significantly to disease management and selection of an appropriate anti-amoebic therapy. Recently, the loop-mediated isothermal amplification (LAMP) method has been applied to the clinical diagnosis of a range of infectious diseases. Here, we describe a rapid and efficient LAMP-based method targeting Acanthamoeba 18S rDNA gene for the detection of Acanthamoeba using clinical ocular specimens in the diagnosis of AK. Acanthamoeba LAMP assays detected 11 different strains including all AK-associated species. The copy number detection limit for a positive signal was 10 DNA copies of 18S rDNA per reaction. No cross-reactivity with the DNA of fungi or other protozoa was observed. The sensitivity of LAMP assay was higher than those of Nelson primer PCR and JDP primer PCR. In the present study, LAMP assay based on directly heat-treated samples was found to be as efficient at detecting Acanthamoeba as DNA extracted using a commercial kit, whereas PCR was only effective when commercial kit-extracted DNA was used. This study showed that the devised Acanthamoeba LAMP assay could be used to diagnose AK in a simple, sensitive, and specific manner.

Cellular and Molecular Pathology of Fungi on Plants Studied by Modern Electron Microscopy

  • Sanwald, Sigrun-Hippe
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.27-53
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    • 1995
  • In plant pathology there is an increasing necessity for improved cytological techniques as basis for the localization of cellular substances within the dynamic fine structure of the host-(plant)-pathogen-interaction. Low temperature (LT) preparation techniques (shock freezing, freeze substitution, LT embedding) are now successfully applied in plant pathology. They are regarded as important tools to stabilize the dynamic plant-pathogen-interaction as it exists under physiological conditions. - The main advantage of LT techniques versus conventional chemical fixation is seen in the maintenance of the hydration shell of molecules and macromolecular structures. This results in an improved fine structural preservation and in a superior retention of the antigenicity of proteins. - A well defined ultrastructure of small, fungal organisms and large biological samples such as plant material and as well as the plant-pathogen (fungus) infection sites are presented. The mesophyll tissue of Arabidopsis thaliana is characterized by homogeneously structured cytoplasm closely attached to the cell wall. From analyses of the compatible interaction between Erysiphe graminis f. sp. hordei on barley (Hordeum vulgare), various steps in the infection sequence can be identified. Infection sites of powdery mildew on primary leaves of barley are analysed with regard to the fine structural preservation of the haustoria. The presentation s focussed on the ultrastructure of the extrahaustorial matrix and the extrahaustorial membrane. - The integration of improved cellular preservation with a molecular analysis of the infected host cell is achieved by the application of secondary probing techniques, i.e. immunocytochemistry. Recent data on the characterization of freeze substituted powdery mildew and urst infected plant tissue by immunogold methodology are described with special emphasis on the localization of THRGP-like (threonine-hydrxyproline-rich glycoprotein) epitopes. Infection sites of powdery mildew on barley, stem rust as well as leaf rust (Puccinia recondita) on primary leaves of wheat were probed with a polyclonal antiserum to maize THRGP. Cross-reactivity with the anti-THRGP antiserum was observed over the extrahaustorial matrix of the both compatible and incompatible plant-pathogen interactions. The highly localized accumulation of THRGP-like epitopes at the extrahaustorial host-pathogen interface suggests the involvement of structural, interfacial proteins during the infection of monocotyledonous plants by obligate, biotrophic fungi.

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