• Title/Summary/Keyword: antibody-antigen reaction

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Development of a Biological Reaction and Measurement Control System for Rapid Detection of the Insecticide Imidacloprid Residues (살충제 Imidacloprid 잔류물의 신속한 측정을 위한 생물반응 및 계측제어 시스템 개발)

  • Lim J. K.;Cho H. K.
    • Journal of Biosystems Engineering
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    • v.30 no.2 s.109
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    • pp.114-120
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    • 2005
  • In this study, a biological reaction and measurement control system was developed to rapidly measure the insecticide imidacloprid residues in agricultural products. The biological reaction part of the system was designed to include micro-pumps and valves for fluid transport, and a polystyrene covet as a reaction chamber. The measurement control part of the system consisted of a photodiode with a light-emitting diode for optical density measurement, and a control microcomputer to implement assay. Signal output was read as the rate of change in optical density at 645 nm. The sensitivity of the system was 2.2 ng/mL ($IC_50$). The system could execute a measurement cycle in about 19 minutes. Research will be continued to develop an automatic sampler fur imidacloprid residues from agricultural products.

Site-directed Immobilization of Antibody onto Solid Surfaces for the Construction of Immunochip

  • Paek, Se-Hwan;Cho, Il-Hoon;Paek, Eui-Hwan;Lee, Haewon;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.2
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    • pp.112-117
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    • 2004
  • The performance of an immuno-analytical system can be assessed in terms of its analytical sensitivity, i.e., the detection limit of an analyte, which is determined by the amount of analyte molecules bound to the capture antibody that has been immobilized onto a solid surface. To increase the number of the binding complexes, we have investigated a site-directed immobilization of an antibody that has the ability to resolve a current problem associated with a random arrangement of the insolubilized immunoglobulin. The binding molecules were chemically reduced to produce thiol groups that were limited at the hinge region, and then, the reduced products were coupled to biotin. This biotinylated antibody was bound to a streptavidin-coated surface via the streptavidin-biotin reaction. This method can control the orientation of the antibody molecules present on a solid surface and also can significantly reduce the possibility of steric hindrance in the antigen-antibody reactions. In a two-site immunoassay, the introduction of the site-directly immobilized antibody as the capture enhanced the sensitivity of analyte detection approximately 10 times compared to that of the antibody randomly coupled to biotin. Such a novel approach would offer a protocol of antibody immobilization in order for the possibility of constructing a high performance immunochip.

Diagnosis of porcine reproductive and respiratory syndrome (PRRS) and its serological survey using the reverse transcription and polymerase chain reaction (RT-PCR) and ELISA (RT-PCR과 ELISA를 이용한 PRRS 진단 및 항체가 조사)

  • Chu Keum-Suk;Han Keu-Sam;Han Jae-Cheol;Song Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.27 no.3
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    • pp.273-280
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    • 2004
  • The studies were performed for the PRRS antigen and antibody detection from breeding farms, artificial insemination(AI) center and growing farms in Jeonbuk province. 1. Specific PRRS primers were successfully amplified ORF6 617bp and ORF7 448 bp on agarose gel. 2. RT-PCR method has been establish by commercial kit and the thermal cycler program consisted of 30 cycles: $95^{\circ}C$ for 30 sec, $45^{\circ}C$ for 30 sec, and $72^{\circ}C$ for 45 sec. 3. The results of PRRS antibody test by ELISA method in AI centers were $6.6\%,\;53.3\%$ and breeding farms $65\%,\;65\%\;and\;38.7\%$, respectively. The serological positive of the antibody in gilt higher than sow. 4. The sero-positive of the PRRS antibody showed average $21\%$ in domestic farms, $56.2\%$ in breeding farms, and $29.9\%$ in AI center.

Immunohistochemical diagnosis of hog cholera with peroxidase-antiperoxidase(PAP) complex method (Peroxidase-antiperoxidase(PAP) 복합체법을 이용한 돼지콜레라의 면역조직화학적 진단)

  • Moon, Oun-gyeong;Cho, Hee-tack;Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.215-221
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    • 1990
  • The present study was intended to use the peroxidase-antiperoxidase method for the identification of hog cholera virus(HCV) in the lymphatic organs of HCV-infected pigs. Sections were incubated with primary antibody (rabbit anti-HCV polyclonal or mouse anti-HCV monoclonal), followed by incubation with linkserum (goat anti-rabbit IgG) in excess and rabbit or mouse PAP complex. The viral antigen was localized mainly in the cytoplasms of lymphoid cells and macrophages. Positive reaction cells were frequently detected in the marginal areas of the germinal centers of the spleens, and also found in the tensils and lymph nodes. The method approved to be highly specific for the identification of the virus and allowed a precise localization of the viral antigen in infected cells.

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Use of the enzyme-linked immunosorbent assay for the detection of toxoplasmosis in swine (ELISA를 이용한 돼지 톡소플라스마병의 조기 진단에 관한 연구)

  • Suh, Myung-deuk;Jang, Dong-hwa;Joo, Hoo-don
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.567-575
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    • 1989
  • This study was conducted to evaluate the possibility of application of a microenzyme-linked immunosorbent assay(micro-ELISA) for the serodiagnosis of specific toxoplasma antibodies in swine sera and this test was performed as a microplate system by coating the polystyrene plates with toxoplasma soluble antigen, incubated serially diluted sera, then added horse radish peroxidase labelled goat anti-swine IgG(r) conjugate followed by o-phenylenediamine as substrate. The color development by enzyme-substrate reaction was determined by the photometric reading [ELISA reader at 490nm (OD)] and visual reading. The soluble antigen was prepared from the tachyzoites in mouse peritoneal cavity. A total of 1,200 swine sera from pig slaughter-house and a total of 116 swine sera from pig breeding station (S-C farm) were tested for the detection of antibodies to Toxoplasma gondii. The results obtained were summarized as follows: 1. The optimal reactions of indirect ELISA for the test sera were determined by the dilution of antigen 1:256 and 1:3,200 of horse radish peroxidase conjugate [anti-swine IgG(r)]. 2. The specific togoplasma antibody(IgG) in pigs infected with Tp artificially were detected as the serum titers of 1:64 or 1:128 at one week postinfection. 3. Of a total of 1,200 swine sera from pig slaughter-house 505 samples of sera were detected as positive (42.1%) and of a total of 116 swine sera from S-C pig breeding station 68 samples of sera as positive (58.6%). 4. The specific antibody(IgG) detection rates against a total of 1,200 test sera from pig slaughter-house were not significant between male (43.1%) and female (40.7%). 5. The indirect ELISA was proved to be a sensitive and specific procedure for the serodiagnosis of swine toxoplasmosis and also evaluated as an effective screening test for the large scale of test samples in laboratory.

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Evaluation of Hypersensitivity Reaction to Dental Materials with a Patch Test (첩포시험을 이용한 치과용 재료의 알러지 평가)

  • Jeon, Hee-Sun;Park, June-Sang;Ko, Myung-Yun
    • Journal of Oral Medicine and Pain
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    • v.25 no.2
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    • pp.167-172
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    • 2000
  • Both immune reaction and hypersensitivity reaction are occurred by the same mechanism, the antigen and antibody reaction. The favorable result of this reaction towards a host is called clinically an immune reaction and the opposite results is called an hypersensitivity reaction. Type IV hypersensitivity reaction is a delayed type which is related to the cellular immune reaction and a contact hypersensitivity is included in this type. Various dental materials such as metal (mercury, nickel, chrome, cobalt), resin and eugenol are etiologic substances. Patch test kit is composed of test substance with a controlled concentration which respond only to a susceptible patient and an aluminum chamber, and etiologic substances for hypersensitivity can be easily and comfortably found just by applying the kit to the patient's skin. In this case report, the patch test was performed to a patients with oral lichen planus and the allergen, restorative material was found. After removal of the matching restoration from the patient's mouth, the symptom was improved.

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Serological Detection of Plant Viruses with latex-test (Latex-Test에 의한 식물 바이러스의 검정)

  • 박은경;김정화;이영근
    • Journal of the Korean Society of Tobacco Science
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    • v.1 no.1
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    • pp.33-38
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    • 1979
  • A simple test using latex-agglutination was developed to detect serologically tobacco mosaic virus (TMV), soybean mosaic virus (SoyMV), and barley stripe mosaic virus (BSMV) from infected Plants. Latex spheres ( 0.81 $\mu$, Difco) were adsorbed with immuno globulin purified by electrophoresis from crude antiserum against viruses. The antibody- sensitized latex suspension was mixed with sap from virus -infected leaves in a glass capillary tube (inner diam. 1mm $\times$ 100 mm length) The mixture, after agitation, was observed under a stereo microscope at low magnification (X20 - X4O), to examine the reaction between antigen (Virus) and its antibody. Flocculation occurred when the reaction was positive.

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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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Hematological Change of Korean Native Goats Experimentally Infected with Bovine Leukemia Virus (Bovine Leukemia Virus에 실험감염된 한국재래산양의 혈액상의 변동)

  • 이필돈;김종호;전무형
    • Korean Journal of Veterinary Service
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    • v.18 no.1
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    • pp.1-21
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    • 1995
  • To elucidate pathogenesis of bovine leukemia virus(BLV) in Korean native goats, the goats experimentally infected with BLV were studied especially for the aspects of infectivity and hematological changes. The experimental goats were examined for 27 months by agar-gel immunodiffusion(AGID) test and syncytium formation assay. During this period, changes of total leucocyte, absolute Iymphocyte and atypical Iymphocyte were examined, and the distribution of surface immunoglobulin ( sIg ) -bearing cells and rosette forming cell (RFC) in the peripheral Iymphocyte were also investigated. By indirect immunofluorescence (IFA) and complement dependent antibody cytotoxicity (CDAC) assay using monoclonal antibody(Mab) against bovine leukosis tumor-associated anti-gen(BL-TAA), changes of BL-TAA positive Iymphocyte in peripheral blood were measured. The results obtained through the experiment were summarized as follows. 1. Antibody titers were measured by AGID using gP51 and P24 antigens. The animals were serologically converted at 2 months post-inoculation(pi) in gP51 antigen, whereas sero-converted at 4 months pi in P24 antigen. In comparison with antibody titers for gP51, P24 antigen showed lower titers throughout the trial period. 2. The peripheral lymphocytes from all of the infected goats, as co-cultivated with F8l cells manifested syncytial formation at 4 months pi. 3. On counting total leucocyte, Iymphocyte and atypical Iymphocyte, two out of four infected goats showed normal distribution, while No 2 of the remaining two revealed temporal and No 3, Persistant increasing number of the cells. 4. The optimal condition of rosette formation of the peripheral Iymphocyte of normal Korean native goats was shown in the sheep erythrocyte treated with 0.1M AET for 30 nun at $37^{\circ}C$. When the Iymphocytes were treated in nylon wool column, the number of sIg-bear-ing cell were increased in the nylon wool adherent cells, but RFC was increased in the non-adherent cells. Of the infected goats, No 2 and No 3 showed significantly increasing number of sIg-bearing cells at 18 months pi. 5. The Iymphocytes of No 2 and No 3 goats reacted positively in IFA using Mab against BL-TAA at 12 months pi and 18 months pi, respectively. In CDAC test, all of four infected goats revealed positive reaction at 24 months pi. The higher positive rates were observed in No 2 and No 3 as compared with the remainders.

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Diagnostic Significance of the Serologic Test Using Antigen of Mycobacterium Tuberculosis for Antibody Detection by ELISA (결핵항원에 대한 혈청학적 검사와 진단적 유용성)

  • Park, Jae-Min;Park, Yeon-Soo;Chang, Yeon-Soo;Kim, Young-Sam;Ahn, Kang-Hyun;Kim, Se-Kyu;Chang, Joon;Kim, Sung-Kyu;Lee, Won-Young;Cho, Shang-Rae
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.2
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    • pp.271-279
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    • 1998
  • Background: Diagnosis by direct microscopy and/or by culture of the Mycobacterium tuberculosis from body fluids or biopsy specimens is "Gold standard". However, the sensitivity of direct microscopy after Ziehl-Neelsen staining is relatively low and culture of mycobacteria is time consuming. Detection of mycobacterial DNA in clinical samples by the polymerase chain reaction is highly sensitive but laborious and expensive. Therefore, rapid, sensitive and readily applicable new tests need to be developed. So we had evaluated the clinical significance of serologic detection of antibody to 38 kDa antigen, which is known as the most specific to the M. tuberculosis complex, and culture filtrate antigen by ELISA in sputum AFB smear negative patients. Method: In this study, culture tests for acid fast bacilli with sputa or bronchial washing fluids of 183 consecutive patients who were negative of sputum AFB smear were performed. Simultaneously serum antibodies to 38 kDa antigen and unheated culture filtrate of M. tuberculosis were detected by an ELISA method. Results: The optical densities of ELISA test with 38 kDa and culture filtrate antigen were significantly higher in active pulmonary tuberculosis cases than in non tuberculous pulmonary diseases (p<0.05), but in patients with active pulmonary tuberculosis, those of the sputum culture positive patients for M. tuberculosis were not significantly different from those of the sputum culture negative cases(p>0.05). In the smear-negative active pulmonary tuberculosis patients, the sensitivity of the ELISA using 38 kDa antigen and culture filtrate was 20.0% and 31.4%. respectively. The specificity was 95.3% and 93.9%. respectively. Conclusion : In active pulmonary tuberculosis but smear negative, the serologic detection of antibody to 38 kDa antigen and culture filtrate by ELISA cannot substitute traditional diagnostic tests and does not have clinically significant role to differenciate the patient with active pulmonary tuberculosis from other with non-tuberculous pulmonary diseases.

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