• Title/Summary/Keyword: enzyme-linked immunosorbent assay(ELISA)

Search Result 742, Processing Time 0.026 seconds

Enzyme-linked Immunosorbent Assays (ELISA) and Immunochromatography Assays (ICG) for Analysis of Vitellogenin in the Scorpion Fish Sebastiscus marmoratus (쏨뱅이(Sebastiscus marmoratus)의 Vitellogenin 분석을 위한 효소면역측정법(ELISA) 및 면역크로마토그래피분석법(ICG) 개발)

  • Yeo, In-Kyu;Lim, Yoon-Kyu
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.48 no.4
    • /
    • pp.459-465
    • /
    • 2015
  • We tested biomarker systems [enzyme-linked immunosorbent assay (ELISA) and immunochromatography assay (ICG) kits] for the screening of endocrine-disrupting chemicals in contaminated environments using antibodies resulting from $17{\beta}$-Estradiol-induced vitellogenin (Vtg) in the wild scorpion fish Sebastiscus marmoratus. Monoclonal antibodies of two clones (S28 and S15) were used as capture and tracer antibodies for ELISA and ICG assays. ELISA detected Vtg at levels greater than $0.1{\mu}g/mL$, while ICG detected Vtg at levels greater than $1{\mu}g/mL$. However, the ICG system was able to detect antibodies from $17{\beta}$-Estradiol-induced Vtg serum that had been diluted 1,000 times. Our results suggest that previously developed biomarker assays can be used as detection systems to detect known endocrine-disrupting chemicals in contaminated environments, and to measure their activity.

Detection of Antibody to Infectious Laryngotracheitis Virus by Enzyme Linked Immunosorbent Assay (효소면역법에 의한 닭 전염성 후두기관염 바이러스 항체 측정에 관한 연구)

  • 임숙경;위성하;최정옥;고홍남
    • Korean Journal of Veterinary Service
    • /
    • v.15 no.1
    • /
    • pp.32-45
    • /
    • 1992
  • In order to establish and enzyme-linked immunosorbent assay to ILTV, field virus strain of ILTV was propagated in chorioallantoic membrane of the embryonated eggs. purified and used as antigen. The antisera selected from the field samples and immunized chickens based on serum neutralization test were used as the standard positive and negative sera in all tests. It was found that optimal antigen concentration was $2{\mu}g$ of protein per well and a 1 : 100 dilution of standard serum showed low background optical density with negative serum and high P/N values of positive sera. A 1 : 500 dilution of the rabbit anti-chicken IgG peroxidase conjugate produced a high P/N values and thirty minutes was chosen as suitable time to read the optical density of the enzyme substrate reaction and optical density was consistent during the 16 hours after stopper was treated. When coated antigen was kept on microplate for varying time up to 16 hours at $4^{\circ}C$ or $37^{\circ}C,$ no significant difference was observed between the treatment. The coated antigen could be kept without change of antigenicity for at least one month at $-70^{\circ}C,\; -20^{\circ}C,\; 4^{\circ}C$ and room temperature. When blocking buffer contanining bovine serum albumin was mixed directly with conjugate and serum at 10% level induced higher P/N values compared to blocking antigen coated microplate with the same blocking buffer. The coefficience of correlation between ELISA and SN test was 0.577. When antibody response of chickens, vaccinated with ILTV, was examined by ELISA and SN test, antibody rising and decay pattern between the two test was similar until 11 weeks of age. However 12 weeks onward antibody titer checked on by SN test was slightly lower than that tested by ELISA.

  • PDF

A Rapid and Sensitive Two-Site Sandwich Enzyme-Linked Immunosorbent Assay for Detection of ${\alpha}$-Fetoprotein in Human Serum

  • Jang, Jeong-Su;Kim, Jeong-Min;Chung, Gi-Hyun;Paik, Bo-Hyun;Kim, Hack-Joo
    • BMB Reports
    • /
    • v.29 no.3
    • /
    • pp.192-199
    • /
    • 1996
  • A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.

  • PDF

Enzyme-Linked Immunosorbent Assay (ELISA) for Detection of Clostridium botulinum Type F Toxin (Clostridium botulinum Type F Toxin의 면역학적 효소방법에 의한 검출에 관한 연구)

  • Lee, Jeong-Kug;K. H. Yang
    • Microbiology and Biotechnology Letters
    • /
    • v.10 no.3
    • /
    • pp.205-209
    • /
    • 1982
  • The enzyme-linked immunosorbent assay using the so-called "double-sandwich"technique was applied to determine Clostridium botulinum type F toxin. Polystyrene tubes were coated with horse anti-type F toxin serum and then toxin sample was added. The tubes were subsequently treated with rabbit anti-type F toxin IgG and sheep anti-rabbit serum IgG-horseradish peroxidase conjugate. By this technique, about 10 mouse intraperitoneal 50% lethal doses (ip LD/50/) of type F toxin could be detected. Low back-ground reading was achieved with the use of phosphate-buffered saline containing 0.05% Tween 20 and 1% bovine serum albumin as diluents of rabbit IgG and conjugate. Addition of EDTA in the diluents of toxin increased ELISA extinction value significantly. No cross-reaction was observed with botulinum type A and B toxin, but type E toxin gave sleight cross-reaction.

  • PDF

Studies on Enzyme-Linked Immunosorbent Assay(ELISA) for Detection of antibody to Mycoplasma hyopneumoniae (돼지의 유행성폐렴 원인균(Mycoplasma hyopneumoniae)에 대한 항체가 분포도 조사)

  • 어용준;육동현;이재문;김윤기;이정학
    • Korean Journal of Veterinary Service
    • /
    • v.22 no.1
    • /
    • pp.9-13
    • /
    • 1999
  • Mycoplasmal pneumonia of swine(MPS) cause by Mycoplasma hyopneumoniae has been recognized as a serious impediment to swine production due to chronic respiratory disorder which result in the weight loss and decreased feed conversion. The disease causes a great economic losses in pig industry by characterizing with high morbidity, low mortality, growth retardation and low feed efficiency. The present study was conducted to investigate the titers of antibody against M hyopneumoniae from the regional and seasonal groups of the slaughtered pigs by enzyme-linked immunosorbent assay(ELISA). The result have shown that the average seropositive rate of M hyopneumoniae infection was 84.6% . The regional seropositive rate in Korea showed 87.4% in Kyonggj, 83.4n in Kangwon, 89.2% in Chungnam and 77.6% in Chungbuk area, respectively. Also the seasonal seropositive rate was appeared as 78.6% in spring,90.1% in summer, 76.9% in autumn and 83.8% in winter, respectively.

  • PDF

Production and Characterization of Monoclonal Antibodies to a Generic Hapten for-Class-Specific Determination of Organophosphorus Pesticides

  • Jang, Mi-Soon;Lee, Soo-Jung;Xue, Xiaoping;Kwon, Hyuk-Man;Ra, Choon-Sup;Lee, Yong-Tae;Chung, Tae-Wan
    • Bulletin of the Korean Chemical Society
    • /
    • v.23 no.8
    • /
    • pp.1116-1119
    • /
    • 2002
  • Monoclonal antibodies have been generated against a generic hapten, ο,ο-diethyl ο-(5-carboxy-2-fluorophenyl) phosphorothioate, for the determination of organophosphorus (OP) pesticides in a class-specific manner. In an indirect competitive enzyme-linked immunosorbent assay (ELISA) format, employing a heterologous coating antigen, these monoclonal antibodies showed desirable properties for use in the class-specific determination, i.e., broad specificity and high sensitivity. The IC50 values of four commonly used ο,ο-diethyl OP pesticides were fairly uniform ranging from 0.1 to 0.3 ㎛/mL. The IC50 values of three ο,ο-dimethyl derivatives were between 0.3 and 1.4 ㎛/mL. These values, together with the limits of detection (LOD), were better, in terms of the specificity and sensitivity, compared with the values obtained previously with polyclonal antibodies.

Seroprevalence of Antigens to Bovine Viral Diarrhea Virus in Korean Calves of the Shown Healthy, Digestive and Respiratory Symptom (한우송아지에서 ELISA를 이용한 소 바이러스성 설사병 바이러스 항원 검출)

  • Chon, Seung-Ki;Park, Jin-Ho;Kim, Nam-Soo
    • Journal of Veterinary Clinics
    • /
    • v.24 no.2
    • /
    • pp.150-153
    • /
    • 2007
  • The aim of this study was to investigate the prevalence of bovine viral diarrhea virus (BVDV) infection in Chonbuk province. Blood samples were taken from 92 korean calves to determined their serological status against BVDV, Capture enzyme-linked immunosorbent assay (ELISA) was used to test for antigen. The number of seropositive calves ranged from 3.3% to 12.9%. Antigens against BVDV were detected in 3.3% of healthy calves, 6.4% of digestive symptom calves, 12.9% of respiratory symptom calves, respectively. Sex and age of calves had no significant differences on the prevalence of BVDV. The results indicate that transmission of BVDV may have become exposed as a result of contact with acute infected or persistently infected cattle.

Enzyme-linked immunosorbent assay for detection of Trichinella spiralis antibodies and the surveillance of selected pig breeding farms in the Republic of Korea

  • Wee, Sung-Hwan;Lee, Chung-Gil;Joo, Hoo-Don;Kang, Yung-Bai
    • Parasites, Hosts and Diseases
    • /
    • v.39 no.3
    • /
    • pp.261-264
    • /
    • 2001
  • Trichinellosis is a parasitic zoonosis of public health importance. It is caused by Trichinella spiralis which has a wide host range including humans. In the present communication, the ELISA technique was employed on a total of 803 blood samples from 7 selected pig breeding farms in 1996 for diagnosis and surveillance of trichinellosis. Out of the entire 803 samples, nine were found to be suspected while one was positive by ELISA. But western blot analyses employed for further confirmation have shown that all of 10 samples did not react to larval excretory-secretory product antigens. These results indicate that pig breeding farms included in the present study are free from trichinellosis . However, it does not mean Korea is free from trichinellosis since human trichinellosis has recently been reported. The necessity of continued surveillance for trichinellosis in both pigs and wild animals was discussed.

  • PDF

Evaluation of the Estrogenic Activity by Yeast Two-hybrid Assay and Enzyme-linked Immunosorbent Assay in Sewage Treatment Plant (하수처리장의 내분비계장애물질에 대한 Yeast Two-hybrid Assay와 Enzyme-linked Immunosorbent Assay에 의한 에스트로겐활성도 평가)

  • Lee, Byoung-Cheun;Ra, Jin-Sung;Kim, Sang-Don;Kawai, Hukiko;Lee, Chul-Hee
    • Journal of Korean Society of Environmental Engineers
    • /
    • v.29 no.7
    • /
    • pp.771-777
    • /
    • 2007
  • Several endocrine disrupting chemicals(EDCs) were monitored to evaluate the estrogenic activities and the concentrations by yeast two-hybrid assay and enzyme-linked immunosorbent assay(ELISA) in sewage treatment plant(STP) which consist of industrial and domestic line. In the influent of domestic line, estrone, 17$\beta$-estradiol, 17$\alpha$-ethinylestradiol and alkylphenolethoxylate(APE) were detected up to 167.1, 39.7, 7.3 and 145.4 ng/L, respectively. The average removal efficiency of 17$\beta$-estradiol after the activated sludge process was 77.5% and further removed to 80.8% after the sand filtration-ozonation step. These results suggests that the activated sludge process has limited potential to remove the estrogenic activity effectively. The contributions of the estrogenic chemicals to the estrogenic activities were 70.7, 23.3, 3.7 and 2.3% for estrone, 17$\beta$-estradiol 17$\alpha$-ethinylestradiol and APE, respectively, in the domestic line effluents. Therefore, 17$\beta$-estradiol and estrone contributed most of the estrogenic activity in the domestic line effluents.

Assay of Cellobiohydrolnse by Column Single Immunodiffusion and Enzyme tinted Immunosorbent Assay (면역화학적 방법에 의한 Cellobiohydrolase 정량)

  • 오태광;고영희;김정일;박관희
    • Microbiology and Biotechnology Letters
    • /
    • v.16 no.3
    • /
    • pp.226-230
    • /
    • 1988
  • Antibody against cellobiohydrolase purified from Trichoderma viride had been obtained by injection to rabbit. The antibody had a high specificity against the cellobiohydroase evidienced by absence of immunological reaction to other isozymes from Trichoderma viride. Assay limit of cellobiohydrolase was 1-10 $\mu\textrm{g}$ by column single immunodiffusion and by enzyme linked immunosorbent assay, it was 10-140 ng and 100-1200 pg when the dilution of antibody was 10$^{-6}$ and 10$^{-5}$, respectively.

  • PDF