• 제목/요약/키워드: antibody-antigen reaction

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항혈청 투여에 따른 돼지 전염성 위장염 예방효과 I. 혈청학적 결과, RT-PCR 검사, 형광항체검사 (Preventive Effects on Transmissible Gastroenteritis(TGE) Using by TGEV Antiserum I. Serological Results, RT-PCR for Fecal and Small Intestin, FA Test)

  • 지영철;한정희;권혁무;한태욱;정현규;박봉균
    • 한국수의병리학회지
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    • 제6권1호
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    • pp.11-18
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    • 2002
  • The purpose of this study was to investigate to potective effects against transmissible gastyoenteritis virus (TGEV) infection in piglets by administration of the TGEV antiserum orally at 5 hrs, 24hrs and 36hrs after birth. five piglets administered the antiserum were experimentally infected with TGEV at four-day-old. Control group were four piglets infected with TGEV only. Serum antibody titers against TGEV were examined by serum neutralization(SN) test, dectection for TGEV or TGEV antigen from feces and small intestines was tested by reverse transcrption-polymerase chain reaction (RT-PCR) and indirect immunoflurescence (IFA). The results obtained were as follows; 1. The piglets administered the TGEV antiserum showed higher antibody titers than those of control group and sustained during the experimental period. 2. The detection rate of TGEV in feces and small intestines by RT- PCR were 24.5% and 20.0% in TGEV antiserum treated group and 44.0% and 75.0% in control group, respectively. 3 The detection rate of TGEV antigen in the small intestine by IFA were 26.7% in TGEV antiserum treated group and 75.0% in control group, respectively. It was concluded that oral administration of antiserum against TGEV to piglets was effective in preventing TGEV infection.

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20(S)-Protopanaxatriol inhibits release of inflammatory mediators in immunoglobulin E-mediated mast cell activation

  • Kim, Dae Yong;Ro, Jai Youl;Lee, Chang Ho
    • Journal of Ginseng Research
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    • 제39권3호
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    • pp.189-198
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    • 2015
  • Background: Antiallergic effect of 20(S)-protopanaxatriol (PPT), an intestinal metabolite of ginseng saponins, was investigated in guinea pig lung mast cells and mouse bone marrow-derived mast cells activated by a specific antigen/antibody reaction. Methods: Increasing concentrations of PPT were pretreated 5 min prior to antigen stimulation, and various inflammatory mediator releases and their relevant cellular signaling events were measured in those cells. Results: PPT dose-dependently reduced the release of histamine and leukotrienes in both types of mast cells. Especially, in activated bone marrow-derived mast cells, PPT inhibited the expression of Syk protein, cytokine mRNA, cyclooxygenase-1/2, and phospholipase $A_2$ ($PLA_2$), as well as the activities of various protein kinase C isoforms, mitogen-activated protein kinases, $PLA_2$, and transcription factors (nuclear factor-${\kappa}B$ and activator protein-1). Conclusion: PPT reduces the release of inflammatory mediators via inhibiting multiple cellular signaling pathways comprising the $Ca^{2+}$ influx, protein kinase C, and $PLA_2$, which are propagated by Syk activation upon allergic stimulation of mast cells.

전북지역에서 발생한 돼지콜레라의 역학적 특성 (Epidemiological characteristics of classical swine fever outbreak at Jeonbuk area in 2003)

  • 엄성심;이정원;서이원;배정준;정동석
    • 한국동물위생학회지
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    • 제27권3호
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    • pp.239-247
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    • 2004
  • Classical swine fever (CSF) was confirmed in 19 herds in Jeunbuk provence (Iksan, Gimje, Wanju, Buan, and Jangsu) in Korea between March and May, 2003 and 10,263 pigs were slaughtered. Pigs contacted with CSF virus in primary outbreak farm show fever, reduced appetite, arched back and chill in company with sever respirative sign and then most infected farms also were observed to fever, reduced appetite, sudden death, and leukopenia (101 pigs). In order to detecting infectious pig with CSF virus, A total of 555 pigs were inspected in 65 herds and blood samples were collected and serological test (ELISA), antigen ELISA, and reverse transcription-polymerase chain reaction (RT-PCR) had been done. Positive rate were $74\%$ (410 pigs) in antibody ELISA, $2\%$ (11 pigs) in antigen ELISA and $33\%$ (182 pigs) in RT-PCR, respectively. As shown that the RT-PCR was useful than the ELISA for determining CSF virus in blood, meat, and other organs.

Immunomodulatory effect of Tinospora cordifolia and Centella asiatica and its modulation on cyclophosphamide challenge

  • Siddiqui, NA;Ali, Mohd;Singh, Shobhna
    • Advances in Traditional Medicine
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    • 제8권4호
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    • pp.380-385
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    • 2008
  • Ethanolic extracts of T. cordifolia and C. asiatica were evaluated for immunostimulatory effect in mice against sheep RBCs as antigen by three models viz. delayed type hypersensitivity reaction, ercent change in neutrophil count and haemagglutination titre. Immunostimulatory effect in the presence of immunosuppressant agent, cyclophosphamide (100 mg/kg, i.p.) was also investigated. T. cordifolia and C. asiatica significantly (p < 0.001, p < 0.05 respectively) enhanced foot pad thickness when measured after 24 hours of sheep RBC antigen challenge. Both the plant materials increased foot pad thickness even after being subjected to immunosuppressant exposure. T. cordifolia revealed enhanced neutrophil counts, while C. asiatica had no significant effect on neutrophil counts. T. cordifolia exhibited significantly (P < 0.01) elevated neutrophil levels even in the presence of cyclophosphamide administration. Both the plants exhibited humoral antibody response, as haemagglutination titre values were significantly high as compared to control. T. cordifolia and C. asiatica could combat immunosuppressant effect of cyclophosphamide (P < 0.01). This suggests that T. cordifolia and C. asiatica can be regarded as biological response modifiers and can be utilized for the development of immunostimulating agent among plant sources.

균사 외피 항체를 이용한 Streptomyces 속 균주들의 혈청학적 유사성 분석 (Serotaxonomical Analyses of Some Streptomyces Strains Using Antibodies against Cell Envelope)

  • 조성기;김재헌
    • 미생물학회지
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    • 제43권2호
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    • pp.137-141
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    • 2007
  • Streptomyces 5균주 간의 유사도를 혈청학적 방법으로 측정하였다. 항원으로는 충분히 생장한 균체의 외피를 Tween 20으로 용출시킨 용액을 사용하였고 항체는 토끼를 면역시켜 얻었다. 면역확산과 ELISA 결과 항원-항체 반응의 정도가 균주간의 분류학적 유사성과 비례적으로 반영되지 않았다. 즉 군집 A에 속하는 균주들 사이의 항원-항체 반응이 낮게 측정된 반면에 군집 F의 Streptomyces Iavendulae와 군집 A의 Streptomyces viridochromogenes가 특이적으로 비교적 높게 나타났다.

Expression of Lily mottle virus Coat Protein and Preparation of IgY Antibody against the Recombinant Coat Protein

  • Yoo, Ha Na;Jung, Yong-Tae
    • 원예과학기술지
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    • 제32권4호
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    • pp.544-549
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    • 2014
  • Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) are the most prevalent viruses infecting lilies in Korea. Leaf and bulb samples showing characteristic symptoms of virus infection were collected in 2012, and 80 field samples were analyzed by reverse transcription polymerase chain reaction (RT-PCR). The infection frequencies were 79% for LMoV, 5% for LSV, and 3% for CMV. The LMoV coat protein gene was amplified and cloned into the pET21d(+) expression vector to develop serological diagnostic tools to detect LMoV. The resulting carboxy-terminal His-tagged coat proteins were expressed in Escherichia coli strain BL21 (DE3) by induction with IPTG. The recombinant proteins were purified using Ni-NTA agarose beads and used as an antigen to produce polyclonal antibodies in laying hens. The resulting egg yolk immunoglobulin (IgY) specifically recognized LMoV from infected plant tissues in immunoblotting assays and had comparable sensitivity to that of a mammalian antibody. In addition, method of immunocapture RT-PCR using this IgY was developed for sensitive, efficient, and rapid detection of LMoV. Based on these results, large-scale bulb tests and detection of LMoV in epidemiological studies can be performed routinely using this IgY. This is the first report of production of a polyclonal IgY against a plant virus and its use for diagnosis.

Development of Chemiluminescence Immunoassay For the Measurement of Serum Thyroxine(T4)

  • Kim, J.B.;Choe, B.K.;Choi, S.H.
    • 한국가축번식학회지
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    • 제11권1호
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    • pp.16-21
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    • 1987
  • We describe a simple, solid-phase chemiluminescence immunoassay for the meausrement of serum T4. An immunoglobulin G fraction of antibody to thyroxine was passively absorbed onto the walls of polystyrene tubes. The labeled antigen was thyroxine-aminobutylethylisoluminol. After the bindings reaction (37$^{\circ}C$ for 1 hour), the solution is removed by aspiration and the antibody-bound fraction was washed once with buffer. Sodium hydroxide (5mol/1,200${mu}ell$) was added and the mixture incubated for 30 minutes at 6$0^{\circ}C$. Luminescence was initiated by oxidation of the label with micropeeroxidase-hydrogen peroxide and the signal of light emission was intergrated for 10 sec. The light yield was inversely proportional to the concentration of T4 in the standard or sample. An evaluation of the method gave the following values sensitivity of calibration curve 7.5$\pm$2.8 nmol/l (mean$\pm$SD). The intra-assay precision (CV%) was 8.9, 7.3 and 5.4. The inter-assay precision (CV%) was 10.2, 8.1 and 7.1. When seum samples were assayed for T4, the results obtained by solid-phase CIA and the conventional RIA agreed well(n=3.5, r=0.954).

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Enzyme-linked immunosorbent assay를 이용한 바이러스성 출혈성 패혈증 바이러스 감염 넙치(Paralichthys olivaceus)의 특이 항체반응 검사 (Detection of Specific Antibodies Against Viral Hemorrhagic Septicemia Virus in Infected Olive Flounder Paralichthys olivaceus Using Enzyme-Linked Immunosorbent Assay)

  • 황지연;장진현;김동준;권문경;서정수;황성돈;손맹현
    • 한국수산과학회지
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    • 제50권5호
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    • pp.547-552
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    • 2017
  • The viral hemorrhagic septicemia virus (VHSV) has an extensive host range, and infects farmed and wild fish inhabiting both freshwater and marine ecosystems. Enzyme-linked immunosorbent assay (ELISA) is highly useful in diagnosing viral hemorrhagic septicemia. However, ELISA shows high, non-specific background reaction with fish antibodies. In this study, we optimized the antigen and antibody concentrations used for detecting specific antibodies in VHSV-infected olive flounder to reduce non-specific binding, and improve the sensitivity of ELISA. The results suggested that OD (optical Density) values were valid when ELISA was performed with $0.1{\mu}g/well$ of virus, involving blocking with blocking buffer (Roth, Roti-Block), 1:300-1:600 dilution with flounder antisera, and 1:1000 dilution with anti-flounder IgM and HRP-conjugated goat anti-mouse IgG for detecting the VHSV antibody in flounder sera. Furthermore, 11 different VHSV strains isolated in Korea from 2012 to 2016 were used to infect the fish. The results showed no correlation between viral pathogenicity and antibody production. This research is a basic study on the application of antibody detection in the diagnosis of viral hemorrhagic septicemia in the olive flounder.

포도상구균에서 분비하는 장내독소 B(SEB)에 대한 재조합 단백질을 이용한 면역특이적 난황항체 생산 (Production of Immunospecific Egg Yolk Antibody with Recombinant Staphylococcal Enterotoxin B (SEB) Protein)

  • 이성;이상래;정경민;김정우
    • 한국가금학회지
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    • 제39권4호
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    • pp.273-278
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    • 2012
  • 본 연구는 식중독은 물론, 아토피성 피부염의 원인물질로 알려져 있는 포도상구균 장내독소 B(Staphylococcal enterotoxin B; SEB)에 대한 특이 난황항체를 개발하고자 실험을 실시하였다. 우선, SEB 유전자를 클로닝한 다음, 대장균 발현시스템을 이용하여 약 30 kDa 정도의 재조합 SEB 단백질을 생산하였다. 재조합 SEB 단백질을 산란계에 2주 간격으로 3회 면역접종을 실시하고, 혈청 및 난황 내 항체가를 측정한 결과, 면역 후 4주경에 항체가가 최고치에 달하였으며, 산란계로부터 획득한 난황항체를 이용한 Western blot 결과, 재조합 SEB 단백질은 물론, 상용화 SEB 단백질과도 특이적으로 반응한다는 것을 규명하였다. 결론적으로, 식중독과 아토피성 피부염 등의 원인물질로 알려진 SEB에 특이적인 난황항체를 생산에 성공하였으며, 이러한 특이적인 난황항체는 식중독 및 아토피성 피부염의 예방 및 치료에 활용 가능할 것으로 사료된다.

판토텐산의 분석을 위한 효소면역측정법 (An Enzyme-Linked Immunosorbent Assay for Detection of Pantothenic Acid)

  • 손동화;박윤식;배근원
    • 한국식품과학회지
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    • 제32권5호
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    • pp.1009-1014
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    • 2000
  • PA를 분석하기 위하여 효소면역측정법을 개발하고자 하였다. Bc방법과 Po방법으로 BSA에 PA를 conjugation하여 각각의 PA-BSA conjugate(PA-BSA[Bc]와 PA-BSA[Po])를 제조하였으며, 이를 토끼에 면역하여 항PA-BSA 항체를 얻었다. 항PA-BSA[Po] 항체를 사용한 ciELISA의 결과에서 경합반응이 제대로 일어나지 않았기 때문에, 식품 속에 있는 PA를 검출하기 위해서 PA-BSA[Po]을 코팅한 후 항PA-BSA[Bc] 항체를 사용하였다. 이 결과에서 PA의 검출한계가 1 ppm인 것을 확인할 수 있었으며, 교차반응을 통해 PA 유도체들에 대해서도 항PA-BSA[Bc] 항체가 PA에 대해 특이성이 매우 강하였다. 또한, MBA의 결과에서는 그 검출한계가 10ppb인 것을 확인할 수 있었다. 분석시료인 계란(109%), 상추(64%), 소간(344%)의 식품시료에 대한 실험에서 상추를 제외하고는 ciELISA는 MBA의 결과와 비교해 볼 때 양호한 결과를 얻을 수 있었다. 그러므로, ciELISA는 MBA보다 분석시간, 교차반응 등의 면에서 장점이 있어 식품 중 PA의 검출에 효과적으로 활용할 수 있을 것이다.

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