• Title/Summary/Keyword: ELISA method

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Application of Competitive ELISA Method for Estimation of Urinary Aflatoxin M1 Level (ELISA 방법을 이용한 요중 아플라톡신 M1 측정)

  • Kim, Yong-Dae;Kim, Heon
    • Journal of Life Science
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    • v.23 no.2
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    • pp.306-310
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    • 2013
  • We compared the efficacy of the competitive ELISA method for measuring the level of urinary aflatoxin M1 (AFM1) with that of the HPLC-fluorescence detector (HPLC-FLD) method. The recovery rate of AFM1 with the ELISA method was 105% (73-124%), and the coefficient of variation of the analysis was 6.85%. The ELISA method showed a 0.20 pg/ml and 0.62 pg/ml limit of detection and limit of quantitation, respectively. In correlation analysis, the two methods showed a very strong and statistically significant correlation (R=0.96, p<0.01). However, in spite of the strong correlation, the ELISA method tended to overestimate the urinary AFM1 concentration compared to the HPLC-FLD method. These results suggest that the competitive ELISA method may be a useful technique for measuring the AFM1 level in high-throughput urine samples, but it needs to be corrected with a regression equation from regression analysis with the HPLC-FLD method.

Quantitative Analysis of Phosphinothricin-N-acetyltransferase in Genetically Modified Herbicide Tolerant Pepper by an Enzyme-Linked Immunosorbent Assay

  • Shim, Youn-Young;Shin, Weon-Sun;Moon, Gi-Seong;Kim, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.681-684
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    • 2007
  • An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE3l-bar) and efficiently purified by $Ni^{2+}$ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: $0.01{\mu}g/ml$) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [$4.9{\pm}0.4{\mu}g/g$ of sample (n=6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT.

Study on the Analysis Method of Ochratoxin A in Cereals by ELISA Method (곡류중 Ochratoxin A의 검색을 위한 면역분석법에 관한 연구)

  • 김동술;정덕화;이용욱
    • Journal of Environmental Health Sciences
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    • v.20 no.3
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    • pp.54-60
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    • 1994
  • We established ELISA method which is more rapid and safe than conventional analytical method, and then analyzed concentration of ochratoxin A in the rices by ELISA method. The results were as follows: 1. Since ochratoxin A molecular does not contain the group for coupling reaction, it was first proposed to have the function of an antigen. As a result of conjugation of ochratoxin A-BSA derivatives, one molecular of BSA was conjugated with 13 moleculars of ochratoxin A. 2. On the basis of established ELISA to apply for immuno analytical method of ochratoxin A, the minimal level of detection by ELISA method was at 0.5 ppb. 3. Rices (42) were collected from homes of Kyoungnam districts through November 1992 to December 1992, as a result of analysis, two of these were positive. Rice samples of R-1 and R-9 represented ochratoxin A levels of 6.0 ng/g and 10 ng/g, respectively.

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Establishment and application of a solid-phase blocking ELISA method for detection of antibodies against classical swine fever virus

  • Cao, Yuying;Yuan, Li;Yang, Shunli;Shang, Youjun;Yang, Bin;Jing, Zhizhong;Guo, Huichen;Yin, Shuanghui
    • Journal of Veterinary Science
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    • v.23 no.5
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    • pp.32.1-32.11
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    • 2022
  • Background: Classical swine fever (CSF) is a severe infectious disease of pigs that causes significant economic losses to the swine industry. Objectives: This study developed a solid-phase blocking enzyme-linked immunosorbent assay (spbELISA) method for the specific detection of antibodies against the CSF virus (CSFV) in porcine serum samples. Methods: A spbELISA method was developed based on the recombinant E2 expressed in Escherichia coli. The specificity of this established spbELISA method was evaluated using reference serum samples positive for antibodies against other common infectious diseases. The stability and sensitivity were evaluated using an accelerated thermostability test. Results: The spbELISA successfully detected the antibody levels in swine vaccinated with the C-strain of CSFV. In addition, the detection ability of spbELISA for CSFV antibodies was compared with that of other commercial ELISA kits and validated using an indirect immunofluorescence assay. The results suggested that the spbELISA provides an alternative, stable, and rapid serological detection method suitable for the large-scale screening of CSFV serum antibodies. Conclusions: The spbELISA has practical applications in assessing the vaccination status of large pig herds.

Development of an Indirect Non-Competitive Enzyme-Linked Immunosorbent Assay for the Detection of Cronobacter muytjensii in Infant Formula Powder (유아용 조제분유 내 Cronobacter muytjensii 검지를 위한 간접 비경합 면역분석법의 개발)

  • Song, Xinjie;Kim, Myunghee
    • The Korean Journal of Food And Nutrition
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    • v.26 no.4
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    • pp.936-944
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    • 2013
  • Cronobacter muytjensii is an important foodborne pathogen as a potential risk in infant formula powder (IFP). To develop a new and sensitive method for the detection of Cronobacter spp. in IFP, an immunoglobulin G (IgG) specific for C. muytjensii (formerly known as Enterobacter sakazakii ATCC 51329) was developed. Further, an indirect noncompetitive enzyme-linked immunosorbent assay (INC-ELISA) was developed by using the anti-C. muytjensii IgG. As a result, this newly developed INC-ELISA method was found very sensitive for C. muytjensii with detection limit of $6.5{\times}10^3CFU/ml$ in pure culture and 1 cell/25 g of IFP. This INC-ELISA method also displayed excellent specificity for C. muytjensii showing no cross-reactivity with other strains of Cronobacter genus and 11 other foodborne pathogenic strains. These results show that the developed INC-ELISA method was very sensitive, efficient, and rapid for the detection of C. muytjensii. Hence, this method could be applied to the development of diagnostic kits for the rapid and easy detection of C. muytjensii.

Detection of Cymbidium Mosaic Virus and Odontoglosum Ringspot Virus by ELISA and RT-PCR from Cultivated Orchids in Korea (ELISA와 RT-PCR에 의한 국내재배난에서 심비디움 모자이크 바이러스와 오돈토글로섬 윤문 바이러스이 검정)

  • 박원목;심걸보;김수중;류기현
    • Korean Journal Plant Pathology
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    • v.14 no.2
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    • pp.130-135
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    • 1998
  • This study was carried out to detect cymbidium mosaic potexvirus (CymMV) and odontoglossum ringspot tobamovirus (ORSV) in cultivated orchid plants in Korea. The standard double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR) were carried out for detection of the viruses in the collected orchid samples. ELISA was suitable for massive-scale diagnostic method for virus detection in orchids. RT-PCR was rapid, time-saving and reliable detective method, and detection limit data showed that RT-PCR was 103 times more sensitive than ELISA. Of the 321 individual orchids representing 5 orchids genera tested by the ELISA, CymMV and ORSV were detected in 15.6% and 22.4%, and mixed infection of the both viruses with 4.9%, respectively. Of the Cymbidium plants tested, cultivated plants showed 52.5% virus infection rate with either CymMV or ORSV and both viruses.

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The survey of Trichinella spiralis infection in finishing pigs using the pepsin-digestion method and ELISA in Korea (조직인공소화법과 ELISA를 이용한 국내 출하돈의 선모충(Trichinella spiralis) 감염실태 조사)

  • Seo, Hunsu;Woo, Gye-Hyeong;Youn, Hee-Jeong
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.269-277
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    • 2004
  • Trichinella spiralis is one of the important zoonotic parasites with a wide variety of vertebrates hosts in nature. The purpose of this study were to analyze ESP(Excretory-Secretory Protein) antigen, to evaluate ELISA for the serological diagnosis of Trichinosis, and to survey T. spiralis infection in finishing pigs using the pepsin digestion method and ELISA in Korea. In the analysis of ESP antigen by SDS-PAGE and Western blot, 4 major bands (70, 55, 52.6, and 49 kDa) were revealed from the ESP antigen. Predilection sites of T. spiralis were the diaphragm, the tongue, masseter muscles, intercostal muscle, and hindlimb in orders in the experimentally infected rats. Sera from 581 swine were tested by ELISA with ESP antigen. The 54 (9.3%) sera were suspected as positive reactors, however, these 54 sera were determined as false positives by the use of Western blotting. This study demonstrated that the ELISA was not suitable for the examination of T. spiralis in pork. The diaphragm muscle samples of 251 finishing pigs were tested by the method of pepsin-digestion for the presence of Trichinella larvae, however, T. spiralis was not detected from the samples. We could not find out T. spiralis infection in pig in Korea pork.

Enzyme-linked Immunosorbent Assays for Antibodies against Chlamydia Pneumoniae Compared with Microimmunofluorescence Test with Patients with Chronic Cough (만성 기침 환자에서 혈청 클라미디아 항체에 대한 ELISA와 microimmunofluorescence 검사의 비교)

  • Lee, Hui Young;Kim, Woo Jin
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.1
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    • pp.47-52
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    • 2005
  • Background : Chlamydia pneumoniae is a clinically important pathogen, the diagnosis of such infection being based mainly on serology. Microimmunofluorescence (MIF) is the current standard diagnostic method, but is subjective and time-consuming, so the authors tested the serology of chronic cough patients using an EILSA method for the Chlamydial antibody, which is a more objective method, and compared the results with those of the standard method. Method : Thirty-five patients, who visited Kangwon National University Hospital between August 2003 and July 2004, were evaluated. A MIF and ELISA tests were used to determine C. pneumoniae antibody titers. Nasopharyngeal aspirates were examined by polymerase chain reaction (PCR). The Spearman rank correlation test was used for data analysis. Results : Sensitivities of ELISA for IgG, IgA and IgM, as judged by MIF, were 84.0, 84.0 and 40.0% and the specificities were 60.0, 60.0 and 96.7%, respectively. Three patients were Chlamydia PCR positive. Conclusion : ELISA can be a useful tool for studying the seroprevalence of Chlamydia pneumoniae. However, further studies will be required prior to its clinical use.

Studies on Enzyme-linked Immunosorbent Assay(ELISA) for Detection of Antibody to Mycobacterium bovis in Serum and Milk (Enzyme-linked Immunosorbent Assay(ELISA)를 이용한 혈청 및 원유 중의 Mycobacterium bovis 항체 검출에 관한 연구)

  • 심항섭;국정희;박병옥;김성열;박유순
    • Korean Journal of Veterinary Service
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    • v.20 no.2
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    • pp.133-142
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    • 1997
  • In order to supplement a diagnostic method for detection of infectious cattle to bovine tuberculosis, performed ELISA for detection of antibody to if bovis in serum and milk. The diagnostic efficacy of the established ELISA was compared with test of the tuberculin skin test for bovine tuberculosis. The positive corresponding rate of serum ELISA and tuberculin skin test showed 84.3%, milk ELISA and tuberculin skin test showed 75.0%, milk ELISA and serum ELISA showed 75.0% respectively. Comparison of the serum and milk to tuberculin antibody concentration in tuberculin positive cattle, the milk contained 1/100-1/150 concentration compared serum tuberculin concentration. The established ELISA was considered efficient for detection of antibodies to M bovis in serum and milk.

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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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