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

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Serodiagnosis of Typhoid Fever by Enzyme-Linked Immunosorbent Assay(ELISA) (효소면역측정법에 의한 장티푸스의 혈청학적 진단)

  • Hwang, Eung-Soo;Cho, Myung-Je;Cha, Chang-Yong;Choe, Kang-Won;Lee, Seung-Hoon;Chang, Woo-Hyun
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.3
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    • pp.387-391
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    • 1986
  • Serum samples from 51 patients with clinically suspected typhoid fever were tested for immunoglobulin G (IgG), IgM and IgA antibodies against the whole bacteria antigen of Salmonella typhi by an enzyme-linked immunosorbent assay. The levels of IgG and IgA antibody to-whole bacteria antigen were higher in the culture-proven patients than in controls. The levels of IgM antibody to- whole bacteria antigen showed better discrimination between culture negative patients and controls than those of IgG or IgA antibody to-whole bacteria antigen. The enzyme-linked immunosorbent assay was much more sensitive than the Widal test. It would be a useful tool for the diagnosis of typhoid fever with a single serum sample.

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An Enzyme-Linked Immunosorbent Assay for Carp (Cyprinus carpio) Vitellogenin and Assay for Oestrogenic Chemicals

  • Jeong, Jing-Woon;Park, Eon-Jung;Kim, Andre;Park, Jang-Su;Park, Heung-Jai
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2001.05a
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    • pp.238-239
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    • 2001
  • The egg-yolk precursor vitellogenin(VTG) is secreted by the liver of female and male fish, in response to estrogenic compound. Carp(Cryprinus carpio) vitellogenin of one major protein is 160kDa, two minor proteins is 110kDa and 170kDa. We were induced vitellogenin by inject of 17-estradiol and purified in a two step procedure by separating it from plasma protein precipitated by 35% saturated ${(NH_4)}_2SO_4$ and then from the remainder by Mono-Q chromatography. We found major carp(Cyprinus carpio) vitellogenin band at 160kDa. Effect of different concentration of oestrogen on vitellogenin synthesis in carp(Cyprinus carpio) exposed for 4 weeks. Show differential effects on vitellogenin synthesis 7 days after treatment. Plasma vitellogenin was measured 3 times for each by an enzyme linked immunosorbent assay(ELISA). ELISA was developed for the detection of the egg yolk precursor vitellogenin in plasma of carp(Cyprinus carpio). The ELISAS performance was optimized and characterized.

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A comparison of agglutination tests and enzyme-linked immunosorbent assay for the bovine brucellosis (소 브루셀라병 혈청 항체가 응집반응 비교 시험)

  • Kim, Cheol-Ho;Bang, Sang-Young;Jeon, Jae-Hyung;Bhak, Jong-Sik;Lee, Min-Kwon;Shin, Jung-Sub
    • Korean Journal of Veterinary Service
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    • v.31 no.3
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    • pp.315-329
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    • 2008
  • A total of 710 bovine serum samples which are composed 532 bovine serum samples showed negative reaction and 178 bovine serum samples showed positive reaction with tube agglutination test (TAT) from North area of Gyeong-nam, Korea were tested using all the 3 assays which are Rose-Bengal test(RBT), tube agglutination test (TAT) and enzyme-linked immunosorbent assay (ELISA, two types) and analyzed for evaluation of specificity, sensitivity, reproducibility and predictive value. In the comparison of serum antibody titer agglutination test, RBT showed almost agreement with TAT. In the comparison of TAT and two types of ELISA method, they showed difference in specificity and sensitivity about 5%. But there is no significant difference in detecting sensitivity between two types of ELISA method and TAT. In serologic tests for bovine brucellosis, the new assay ELISA would be a good candidate for serologic survey for bovine brucellosis in Korea because it is efficient in detecting many test samples quickly. But the serum agglutination tests (RBT, TAT) are more economical and easy assay for detection. In the test of comparison of antibody titer between first day of finding and 10 days after finding by TAT, there was no change in 55% (76/139) of positive cattle.

Enzyme-Linked Immunosorbent Assay for Detection of Nivalenol (Nivaleno의 검출을 위한 효소 면역 측정법)

  • 손동화;이향범;곽보연;김수호;권창희
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.129-134
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    • 1998
  • To develop an enzyme-linked immunosorbent assay (ELISA) for nivalenol (NIV), we produced polyclonal antibodies against tetraacetyl nivalenol (Ac4-NIV) and established ELISA conditions. Ac4-NIV-hemisuccinate conjugated to bovine serum albumin (Ac4-NIV-HS-BSA) was immunized with Freund's adjuvants into rabbits subcutaneously several times. By use of the antiserum showing the highest titer and Ac4-NIV-HS-HRP conjugate, we established competitive direct ELISA (cdELISA). Standard curve of cdELISA showed that the detection range of Ac4-NIV was about 10~5,000 ng/ml (ppb). The cross-reactivities of the polyclonal antibody towards Ac4-NIV and acetyl T-2 were 100 and 70% respectively, and those towards NIV, deoxynivalenol, 3-acetyl deoxynivalenol, 15-acetyl deoxynivalenol, triacetyl deoxynivalenol, fusarenonX, and T-2 were less than 0.1%. When cdELISA was applied to NIV-spiked corns followed by extraction with 70% acetonitrile and acetylation with acetic anhydride in pyridine, the recovery rates of the Ac4-NIV were 108, 143, and 70% (average, 107%) in the levels of 100, 300, and 1,000 ng/g (ppb), respectively.

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Development of a blocking ELISA for detection of Japanese encephalitis virus antibodies in pig and horse sera

  • Dong-Kun Yang;Eun-Ju Kim;Sang Ho Jang;Hye Jung Lee;Bitna Kim;Jin A Lee;Ju-Yeon Lee;Yun Sang Cho
    • Korean Journal of Veterinary Research
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    • v.64 no.3
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    • pp.26.1-26.9
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    • 2024
  • Japanese encephalitis virus (JEV) is a mosquito-borne virus that can infect pigs, horses, and other mammals, including humans. Sero-epidemiological investigations of JEV have been performed using hemagglutination inhibition (HI), virus neutralization (VN) tests and enzyme-linked immunosorbent assay (ELISA). A need exists for a new ELISA that can detect JEV antibodies in the sera of several animal species. We aimed to develop a blocking ELISA (B-ELISA) for detecting JEV antibodies in pig and horse serum samples. JEV antibodies in 218 pig and 315 horse serum samples were measured using HI and VN tests. The purified KV1899-306 strain was used as an antigen for B-ELISA. The purified antibody (7A13) was conjugated with horseradish peroxidase and used as a detector antibody. The sera of pigs and horses to measure antibody against JEV were subjected to B-ELISA and analyzed. The B-ELISA had a diagnostic sensitivity of 94.6% to 100%, a specificity of 91.2 to 100%, and an accuracy of 94.9 to 98.6% compared with those of the HI and VN tests in pig and horse sera. The B-ELISA had a higher correlation with pig sera (r = 0.89 and 0.90 for VN and HI) than with horse sera (r = 0.75 and to 0.79). The new B-ELISA could be useful in the sero-surveillance of JEV in pig and horse sera and replace indirect ELISA.

An Enzyme-Linked Immunosorbent Assay for Detection of Milk proteins in Food (우유단백질의 분석을 위한 효소면역측정법)

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Bae, Gun-Won;Kim, Soon-Mi
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.564-569
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    • 2000
  • An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of milk proteins in processed foods. The ${\alpha}_{s1}-casein({\alpha}_{s1}-CN)$, a heat stable major milk protein, was immunized into rabbits to produce specific antibodies. When competitive indirect ELISA(ciELISA) using $anti-{\alpha}_{s1}-CN$ antibodies was established, its detection limit was $0.1\;{\mu}g/mL$. The reactivities of the specific antibodies toward ${\alpha}_{s1}-CN$, skim milk, ${\beta}-CN$ and whey protein isolate(WPI) were 100, 37, 0.14 and 0.04%, respectively, as determined by ciELISA. However $anti-{\alpha}_{s1}-CN$ antibodies did not have any reactivity to other milk proteins such as ${\beta}-lactoglobulin,\;{\alpha}-lactalbumin$, bovine serum albumin, and isolated soy protein. When sandwich ELISA was established, its detection limit was $0.01\;{\mu}g/mL$ which was 10 times more sensitive than that of ciELISA. In the spike test which was performed by adding 1-10% of whole CN to market milk, mean assay recovery as determined by sandwich ELISA was 94.8%(CV, 8.2%). Food stuffs and dairy products were assayed by sandwich ELISA to show 29, 0.13, 0.25, and 6.9% of whole CN in skim milk powder, WPI, semi-solid yoghurt, and processed cheese, respectively.

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An Enzyme-Linked Immunosorbent Assay for Quantitation of Soy Proteins in Food (식품 중 대두단백질의 정량분석을 위한 효소면역측정법)

  • Shon, Dong-Hwa;Kim, Hyung-Jung;Eum, Byong-Wook;Kim, Soo-Ho;Kim, Soon-Mi
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.991-996
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    • 2000
  • Enzyme-linked immunosorbent assay was developed for the analysis of soy protein in foods. Competitive indirect ELISA (ciELISA) was established by using specific antibodies against the heat-stable acidic subunits (AS) of glycinin. Soy proteins in each sample used in this study were solublized in the presence of urea and DTT and boiled at $100^{\circ}C$ for 1hr and then were renatured with a cystine-containing solution. After these treatments, each isolated soy protein (ISP) heated at 60, 70, 80, $90^{\circ}C$ for 10 minutes showed almost the same curve as unheated one in the ciELISA. The detection limit of ISP was 0.3 ${\mu}g/mL$. Anti-AS antibodies have very low reactivities less than 0.1% toward non-meat proteins such as skim milk and casein and did not show any reactivities toward egg white powder and ovalbumin. When laboratory-made sausages containing ISP of $0.5{\sim}3%$ were assayed by ciELISA, the mean recovery was about 83% (C.V., 19%). In addition, the average content of soy protein in commercial sausages was 1.27%.

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A study on analytical methods for polycyclic aromatic hydrocarbons in foods (식품 중 다환방향족탄화수소 분석법 연구)

  • Kim, Yong-Yeon;Shin, Han-Seung
    • Food Science and Industry
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    • v.55 no.1
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    • pp.45-57
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    • 2022
  • This study was proceeded the analytical methods using various analytical instruments for polycyclic aromatic hydrocarbons (PAHs) in food products. Various analytical methods were developed to determine levels of PAHs including benzo[a]pyrene, benzo[a]anthracene, benzo[b]fluoranthene, and chrysene formed in various food products using gas chromatography-mass spectrometry (GC-MS), enzyme-linked immunosorbent assay (ELISA) and raman spectroscopy. Recently, the rapid on-site response for the detection of hazardous substances in food aims to develop an onsite rapid detection of a simplified technical analysis method to reduce the time and cost required for analysis of PAHs. Current PAHs detection methods have been reviewed along with new raman spectroscopy analytical method.

Comparison of TIA with ELISA for circulating antibody detection in clonorchiasis (간흡충증에 있어서 항체검출을 위한 Enzyme-linked Immunosorbent Assay와 Thin Layer Immunoassay의 비교)

  • 이용기;유재숙이근태정경일
    • Parasites, Hosts and Diseases
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    • v.21 no.2
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    • pp.265-269
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    • 1983
  • A comparison was made of a new serological method, thin layer immunoassay (TIA), and an established method, enzyme-linked immunosorbent assay (ELISA), in the detection and quantiacation of antibodies in clonorchiasis. Saline extract of Iyophilized Clonorchis sinensis adult worm was used as antigen, and TIA by the method of Elwing et at. (1976) and ELISA by Voller et at. (1974) were performed. Using sera from known clonorchiasis cases,100% of the sera tested were Positive by TIA and 88.35 by ELISA. TIA produced false positive results in 14 out of 36 cases, which were 10 amoebiasis cases, 16 paragonimiasis cases and 10 healthy controls. ELISA. however, produced a small number of false positives, 7 out of 55 cases. There was correlation between Immunoglobulin G level in sera and ELISA value (correlation coefficient, 0.69), whereas no correlation between Immunoglobulin G level and TIA result. The Performance of TIA and ELISA was not correlated in the results using homologous antigen.

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Standardization of an enzyme-linked immunosorbent assay for detection of antibody to avian reticuloendotheliosis virus (세망내피증 바이러스 항체검출을 위한 ELISA 표준화)

  • Sung, Haan Woo;Lee, Su Jeong
    • Korean Journal of Veterinary Research
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    • v.45 no.4
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    • pp.569-574
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    • 2005
  • Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to reticuloendotheliosis virus (REV) at single serum dilution was standardized. REV HI, one of the Korean field isolates, was inoculated into chicken embryo fibroblast (CEF) cells and was harvested from the culture fluids and cells after 10 to 12 days. Viruses were purified by centrifugation at the $107,000{\times}g$ for 12 hours on 20, 30, 45% (W/V) sucrose gradient. Virus specific fraction was collected and used as ELISA antigen. To standardize ELISA, the optimal concentration of coating antigen ($1{\mu}g/well$) and conjugate (1/1000) was determined by corrected OD (OD value of positive serum-OD value of negative serum) and P/N ratio (OD value of positive serum/OD value of negative serum). To calculate ELISA titer by measuring absorbance at 1/400 single serum dilution, serum titrations were carried out for various sample sera together with standard positive and negative sera. The observed titers of serum samples were plotted against sample/positive (s/p) ratios at 1/400 serum dilution. From the above data, the ELISA titers could be calculated by the equation of $log_{10}$ ELISA titer = 2.2763 ($log_{10}$ s/p) + 3.482 (r = 0.93). For evaluating the sensitivity, the standardized method were compared with conventional agar gel immunodiffusion (AGID) test method using serum samples collected from REV infected field chicken flocks. Fifty seven of 60 samples (95%) were positive for REV by ELISA, whereas only 11 (18.3%) samples were positive by AGID test. This results suggested that the ELISA tests developed in this study could be used for detection of antibodies to REV with high sensitivity.