• Title/Summary/Keyword: Specific reactivity

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Properties of Anti-S. mutans IgY Separated from Egg Yolk (난황으로 부터 항충치 항체의 분리 및 그 특성)

  • Shon, Dong-Hwa;Rho, Jeong-Hae;Kim, Young-Boong;Han, Chan-Kyu;Sung, Ki-Seong;Lee, Nam-Hyung
    • Korean Journal of Food Science and Technology
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    • v.30 no.5
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    • pp.1029-1034
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    • 1998
  • Chick antibodies (IgY) raised against Streptococcus mutans (serotype c) were separated from egg yolk and their properties were investigated. The purity of IgY extracts prepared by the method of ${\lambda}-carrageenan$, $gammaYolk^{TM}$, and $EGGstract^{TM}$ was 20%, 46%, and 48%, respectively, and the yields of IgY extracts from a gram yolk were 11. 3 mg, 1.7 mg, and 1.8mg, respectively. Quantitative immunoprecipitation test showed that specific IgY content of crude IgY prepared by ${\lambda}-carrageenan$ method was 12.2%, which means that 0.85 g of crude IgY from an egg yolk (15 g) contains about 100 mg of specific IgY. When the reactivity of the specific IgY towards 3 caries-inducing strains (serotype: b, c, f) was examined, the strains cultured in sucrose-added medium showed higher reactivity (the orders were c(+), f(+), b(+)) than those cultured in sucrose-free medium. Heat and pH stability of specific IgY was good, for crude IgY contained 50% of antibody activity after heat treatment at $70^{\circ}C$ for 5 min and they were stable at pH $4{\sim}8$.

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Analysis of Culture Filtrate Antigens of Aspergillus fumigatus Strains and of Antibody Responce in Patients with Aspergillosis by Immunoblotting (Immunoblotting 에 의한 Aspergillus fumigatus 균주(菌株)의 항원분석(抗原分析)과 이 균(菌)에 감염(感染)된 환자의 항체반응(抗體反應)에 관한 연구)

  • Kim, Sang-Jae;Kim, Sin-Ok;Hong, Young-Pyo
    • The Korean Journal of Mycology
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    • v.17 no.2
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    • pp.66-75
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    • 1989
  • Heterogeneity in antigenic composition of Aspergillus fumigatus isolates from clinical specimens and in antibody response of patients infected with this fungus was investigated by immunoblotting. A considerable quantitative and qualitative difference was found in composition of the culture filtrate antigens derived from a reference strain (ATCC 13073) and 8 clinical isolates of A. fumigatus on SDS-PAGE and immunoblots. The crude CF antigen of a strain AFG7 was selected to identify the serologically reactive and specific components by immunoblotting. Out of more than 36 components separated by electrophoresis, transblotted to nitrocellulose sheet, and reacted with sera that showed a positive reaction to A. fumigatus or other fungal antigens on immunodiffusion tests, merely four or so were found useful to serodiagnosis of aspergillosis. An antigen of 82KD was found most reactive and specific component so as to be contained in the standard preparation. Several other components, for example 11KD, 26KD, 30KD and 31KD, also possessed relatively high reactivity and specificity and seemed to be worth while purifying and characterizing. Antibody binding activity (reactivity) of the antigenic components was clearly shown on immunoblots because some were faintly stained with Coomassie blue but darkly stained on immunoblots, while some others behaved contrary to them. A number of components seemed to carry not only species specific but cross reactive antigenic determinants. Immunoblotting proved very useful to identify serologically reactive and specific components that should be present in the antigen to be employed to the serodiagnosis of aspergillosis.

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Identification of the Chicken Meat Allergens (닭고기 중 알레르기 유발성분의 동정)

  • 조은득;김동섭;정기화
    • Biomolecules & Therapeutics
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    • v.9 no.1
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    • pp.7-14
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    • 2001
  • The chicken meat has been reported as one of the food causing allergic reactions predominantly to Korean. At present, several in vitro tests for immunoglobulinG (IgG)-mediated as well as IgE-mediated food allergy are available. 13 clinically chicken meat-allergic patients were investigated together with 4control subjects for identification of chicken meat-specific reactivity by ELISA. Also, protein profile and IgE, IgGtotal and IgG4-reacting allergens were detected by means of sodium dodecyl sulfate-polyacrylamide gel electro-phoresis (SDS-PAGE)and immunoblotting. Chicken meat extracts were prepared as raw, heated, heat and simulated gastric fluid (SGF) treated samples to characterize the stability of allergen to physicochemical treatment. SDS-PAGE revealed 9~200 kDa bands. And in immunoblotting 7 sera were identified most major bands between 10 and 78 kDa. In case of IgE, six proteins (17, 26, 35, 40, 78 kDa) were predominant in heat-treated extract, and the one (35 kDa) was present in SGF-treated preparations. In case of IgG$_{total}$ and IgG4, most of them showed a patters simmilar to IgE. There were significant differences (P<0.05) in IgE, IgG$_{total}$ , IgG4 Abs to chicken meat between the allergic and control subjects in ELISA. In addition, the concentration of IgG4Abs in the challenge-positive subjects was significantly higher than that of control subjects. It is considered that the specific IgE response to chicken meat was rarely prevalent to Koreans. However, the specific IgG4 response play an important role in the development of allergic symptoms.

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Evaluation of Rapid IgG4 Test for Diagnosis of Gnathostomiasis

  • Wang, Yue;Ma, An;Liu, Xiao-Long;Eamsobhana, Praphathip;Gan, Xiao-Xian
    • Parasites, Hosts and Diseases
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    • v.59 no.3
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    • pp.257-263
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    • 2021
  • Human gnathostomiasis is a parasitic disease caused by Gnathostoma nematode infection. A rapid, reliable, and practical immunoassay, named dot immuno-gold filtration assay (DIGFA), was developed to supporting clinical diagnosis of gnathostomiasis. The practical tool detected anti-Gnathostoma-specific IgG4 in human serum using crude extract of third-stage larvae as antigen. The result of the test was shown by anti-human IgG4 monoclonal antibody conjugated colloidal gold. The sensitivity and specificity of the test were both 100% for detection in human sera from patients with gnathostomiasis (13/13) and from healthy negative controls (50/50), respectively. Cross-reactivity with heterogonous serum samples from patients with other helminthiases ranged from 0 (trichinosis, paragonimiasis, clonorchiasis, schistosomiasis, and cysticercosis) to 25.0% (sparganosis), with an average of 6.3% (7/112). Moreover, specific IgG4 antibodies diminished at 6 months after treatment. This study showed that DIGFA for the detection of specific IgG4 in human sera could be a promising tool for the diagnosis of gnathostomiasis and useful for evaluating therapeutic effects.

Pre-existing Immunity to Endemic Human Coronaviruses Does Not Affect the Immune Response to SARS-CoV-2 Spike in a Murine Vaccination Model

  • Ahn Young Jeong;Pureum Lee;Moo-Seung Lee;Doo-Jin Kim
    • IMMUNE NETWORK
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    • v.23 no.2
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    • pp.19.1-19.10
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    • 2023
  • Endemic human coronaviruses (HCoVs) have been evidenced to be cross-reactive to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Although a correlation exists between the immunological memory to HCoVs and coronavirus disease 2019 (COVID-19) severity, there is little experimental evidence for the effects of HCoV memory on the efficacy of COVID-19 vaccines. Here, we investigated the Ag-specific immune response to COVID-19 vaccines in the presence or absence of immunological memory against HCoV spike Ags in a mouse model. Pre-existing immunity against HCoV did not affect the COVID-19 vaccine-mediated humoral response with regard to Ag-specific total IgG and neutralizing Ab levels. The specific T cell response to the COVID-19 vaccine Ag was also unaltered, regardless of pre-exposure to HCoV spike Ags. Taken together, our data suggest that COVID-19 vaccines elicit comparable immunity regardless of immunological memory to spike of endemic HCoVs in a mouse model.

Effects of Cyclooxygenase Inhibitors on Vascular Reactivity and Alterations of Cyclooxygenase Expression (혈관 반응성에 대한 Cyclooxygenase 억제제 효과와 Cyclooxygenase 발현 변화)

  • Lee, Ki-Young;Park, Jin-Woo;Eum, Eun-A;Kang, Young-Jin;Lee, Kwang-Youn;Choi, Hyoung-Chul
    • Journal of Yeungnam Medical Science
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    • v.23 no.1
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    • pp.36-44
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    • 2006
  • Background: There is controversy regarding whether COX-2 specific inhibitors are associated with elevation of blood pressure. We compared the effects of aspirin, indomethacin, and celecoxib for vascular reactivity induced by phenylephrine. We also tested the effects of indomethacin and NO donor on COX-1 and COX-2 protein expression, as well as nitrite production in culture medium of vascular smooth muscle cells. Materials and Methods: In this experiment, we used the isometric tension study for vascular reactivity. After 45 minutes of pretreatment with aspirin, indomethacin, celecoxib, and phenylephrine induced contractions were tested. COX-1 and COX-2 protein expressions were analyzed by Western blot and nitrite production by the Griess reaction. Results: Although celecoxib pretreatment caused enhanced arterial contraction, aspirin pretreatment induced more potent arterial contraction than celecoxib in the isometric tension study of rabbit femoral artery. COX-1 protein expression was unchanged by indomethacin, SNP and NOR-3; COX-2 protein expression was increased by the addition of indomethacin, SNP, and NOR-3. Especially, NOR-3, a NO donor, significantly increased COX-2 protein expression with unstimulated conditions as well as LPS stimulation. Induction of nitrite production was higher with NOR-3 treatment than SNP treatment with LPS stimulation. Conclusion: These results suggest that aspirin caused more potent vascular contraction than celecoxib and indomethacin. COX-2 expression in VSMC depended on the types of NO donor and LPS stimulation.

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Detection of Serum IgE Specific to Mite Allergens by Immuno-PCR

  • Lee, Kyung-Woo;Hur, Byung-Ung;Chua, Kaw-Yan;Kuo, I-Chun;Song, Suk-Yoon;Cha, Sang-Hoon
    • IMMUNE NETWORK
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    • v.8 no.3
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    • pp.82-89
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    • 2008
  • Background: Although a skin test is the primary option for detecting allergen-specific IgE in clinics, the serum IgE immunoassay is also important because it allows for the diagnosis of allergy without any accompanying adverse effect on the patient. However, the low detection limit of IgE levels by immunoassay may restrict the use of the method in some occasions, and improving its sensitivity would thus have a significant implication in allergy-immunology clinics. Methods: In this study, we attempted to detect specific serum IgE by using immuno-polymerase chain reaction (IPCR) which combines the antigen-antibody specificity of enzyme-linked immunosorbent assays (ELISAs) with the amplification power of PCR. Results: Our results demonstrated that Blo t5-specific serum IgE can be detected by IPCR with a 100-fold higher sensitivity than ELISA, and cross-reactivity of serum IgE to other mite allergens is able to be analyzed by using only $0.3{\mu}l$ of serum sample. Use of real-time IPCR seemed to permit more convenient determination of specific serum IgE as well. Conclusion: We believe that IPCR can serve as a valuable tool in determining specific serum IgE, especially when the amount of serum sample is limited.

A Study on Low-Temperature Oxidation Reactivity of Pt/ZrO2·SO42-Catalyst (ZrO2·SO42-에 담지된 백금촉매의 저온산화반응성에 대한 연구)

  • Kim, Kiseok;Lee, Tae Jung;Kim, Byoung Sam;Kim, Du Soung
    • Applied Chemistry for Engineering
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    • v.9 no.1
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    • pp.141-148
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    • 1998
  • Reactivity of Pt catalysts(0.2, 0.5 wt% Pt) supported on solid super acid, $ZrO_2$ $SO_4{^{2-}}$ for low-temperature oxidation was investigated for complete oxidation of cyclohexane. Catalytic activity measured as reactant conversion in a packed-bed tubular reactor increased in accordance with the acidity and specific surface area of the catalyst activity and specific surface area of $Pt/ZrO_2$ $SO_4{^{2-}}$ catalyst were diminished by adding potassium during catalyst preparation. the catalyst activity decreased in accordance with the amount of potassium added. In addition, $Pt/ZrO_2$ $SO_4{^{2-}}$ catalyst exhibited an activity greater than that of a $Pt/SiO_2$ or $Pt/Al_2O_3$ catalyst possessing much larger specific surface area at $250^{\circ}C$ for the reactant stream of 15.000 ppm cyclohexane concentration and $18,000hr^{-1}$ space velocity, a cyclohexane conversion as high as 96% was obtained over 0.2 wt% $Pt/ZrO_2$ $SO_4{^{2-}}$, whereas cyclohexane conversions over 0.2 wt% $Pt/SiO_2$ and 0.2 wt% $Pt/Al_2O_3$ were 83 and 79%, respectively.

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A Lateral Flow Immunoassay Kit for Detecting Residues of Four Groups of Antibiotics in Farmed Fish (어류 중 4계열 잔류 항생물질 검출을 위한 Lateral Flow Immunoassay Kit 개발)

  • Jo, Mi Ra;Son, Kwang Tae;Kwon, Ji Young;Mok, Jong Soo;Park, Hong Jae;Kim, Hyun Yong;Kim, Gyung Dong;Kim, Ji Hoe;Lee, Tae Seek
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.2
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    • pp.158-167
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    • 2015
  • A lateral flow immunoassay kit based on antigen-antibody interactions was developed to detect residues of beta-lactams, quinolones, tetracyclines, and sulfonamides in farmed fish. Group-specific antibodies showing cross-reactivity with other antibiotics in the same group were produced in rabbits. The rabbits were immunized eight times to obtain the maximum titers. Antibodies were extracted from the antisera collected from the immunized rabbits and produced group-specific reactions with antibiotics from the four groups. A kit was prepared that optimize conditions for the antigen-antibody reaction, using colloidal gold conjugated antibodies, and was designed to detect the four groups of antibiotics simultaneously. The kit enabled the detection of antibiotics in the four groups at below maximum residue limits (MRLs), which were $200{\mu}g/kg$ for tetracyclines, $100{\mu}g/kg$ for sulfonamides, $50{\mu}g/kg$ for beta-lactams, and $100{\mu}g/kg$ for quinolones. The cross-reactivity of the antibodies ranged from 10-80% for the sulfonamides, 20-100% for tetracyclines, 38-100% for quinolones, and 20-100% for the beta-lactams, confirming that the antibodies were group specific. The test kit was used 30 times to examine spiked antibiotics at the limits of detection (LODs) and all produced positive results, indicating high sensitivity. The LODs for the assay ranged from 4-20 ng/mL for beta-lactams, 25-50 ng/mL for sulfonamides, 20-100 ng/mL for tetracyclines, and 30-80 ng/mL for quinolones, and there were no false negative reactions at above these LODs. In addition, all of the LODs of the developed kit were correlated with high-performance liquid chromatography (HPLC) data. Our lateral flow immunoassay kit can simultaneously detect antibiotic residues from a large number of fish samples rapidly, strengthening the safety of domestic farmed and imported fish.

Development of TaqMan probe-based real-time PCR for rapid identification of beef, pork and poultry meat (소, 돼지, 가금육류의 신속한 동정을 위한 TaqMan probe를 이용한 real-time PCR 개발)

  • Koh, Ba-Ra-Da;Kim, Ji-Yeon;Na, Ho-Myung;Park, Seong-Do;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.35 no.3
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    • pp.215-222
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    • 2012
  • Species-specific $TaqMan^{(R)}$ probe-based real-time PCR assays were developed for detection of beef, pork, chicken, duck, goose and turkey. The primer and probe sets used in this study were designed to be complementary to fibroblast growth factor (FGF) for cattle and pig, mitochondrial NADH dehydrogenase (ND) subunit 3 and ND2 for chicken and duck, 12S rRNA for goose and turkey, respectively. As internal positive control we used conserved region in the ribosomal 18S RNA gene to ensure the accuracy of the detection of target DNA by real-time PCR. We confirmed that real-time PCR assays with the primer and probe sets were positive for cattle, pig and chicken intended target animal species with no cross-reactivity with other non-target animal species. Only >50 ng DNA of beef show cross-reactivity in the determination of duck. Using species-specific primer and probe sets, it was possible to detect amounts of 0.1 ng DNA of cattle and pig, 1.0 pg DNA of chicken, duck and turkey, and 0.1 pg DNA of goose for raw samples, respectively. The detection limits were 0.1 ng DNA of cattle, 1.0 ng DNA of pig and 1.0 pg DNA of chicken for DNA mixtures (beef, pork and chicken) extracted from heat-treated ($121^{\circ}C$/5 min) meat samples. In conclusion, it can be suggested that the $TaqMan^{(R)}$ probe-based assay developed in this study might be a rapid and specific method for the identification of meat species in raw or cooked meat products.