• Title, Summary, Keyword: ELISA

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Detection of Salmonella in Milk by Sandwich ELISA using Anti-Outer Membrane Protein Immunoglobulins (Anti-Outer Membrane Protein 면역단백질을 이용한 Sandwich ELISA 방법에 의한 우유 내 Salmonella의 검출)

  • 최석호
    • Food Science of Animal Resources
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    • v.24 no.2
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    • pp.176-181
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    • 2004
  • The specificity of sandwich enzyme-linked immunosorbent assay (ELISA) to detect Salmonella in milk was determined in this study. The antibodies used in sandwich ELISA were egg yolk immunoglobulin G (IgY) obtained after immunization of hen with outer membrane protein (OMP) fraction from Salmonella typhimurium and rabbit IgG obtained after immunization of rabbit with the purified OMP with the molecular weight of 40,000. The immunoblot assay showed that the IgY reacted strongly with OMP with the molecular weight of 6,000 and the rabbit IgG reacted strongly with OMP with the molecular weights of 40,000, 35,000, and 6,000 from the bacteria including Salmonella which belongs to Enterobacteriaceae. The IgY and rabbit IgG also reacted with other proteins from Salmonella typhimurium in immunoblot assay. Competitive ELISA showed that IgY showed specifity to react with two strains of Salmonella typhimurium and Salmonella cholerasuis but not with Escherichia coli and Yersinia enterocolitica. Two strains of Salmonella typhimurium added to UHT milk showed the highest absorbance of all the bacteria used in the sandwich ELISA. Some strains of Salmonella cholerasuis showed higher absorbances than non-Salmonella bacteria.

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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Use of ELISA for the Residue Analysis of Pesticides (ELISA 기법을 이용한 농약(農藥)의 잔류분석(殘留分析))

  • Lee, Kang-Bong;Suh, Yong-Tack
    • Korean Journal of Environmental Agriculture
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    • v.12 no.3
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    • pp.298-308
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    • 1993
  • Immunochemical assay, ELISA for small molecules such as pesticides are rapid, sensitive, cost effective and can easily analyze with large samples. ELISA is one of several powerful biotechnologies immediately applicable to pesticide analysis. This review lists the advantages and disadvantages of the ELISA and elucidate the steps in assay development using examples from this laboratory. The focus is primarily on hapten synthesis strategies, protein conjugation, Immunization, assay format, and assay validation.

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Detection of IgG and IgM antibodies with ELISA technique in human trichomoniasis (질트리코모나스 환자에서 효소표식 면역검사법을 이용한 혈청 내 항-질트리코모나스 IgG 및 IgM 항체가의 측정)

  • 이미리;신명헌
    • The Korean Journal of Parasitology
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    • v.28 no.1
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    • pp.25-30
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    • 1990
  • The direct wet mount examination of vaginal. secretion, widely applied for the diagnosis of Trichcmonas vaginalis infection in woman patients, is rapi4 and economical, however, the sensitivity of this technique is not so high. In this study enzyme-linked immunosorbent assay (ELISA) was employed for the detection of serum anti-T. vaginalis IgG and IsM antibodies from 30 vaginal trichomoniasis patients and 30 non-infected healthy persons. The results were as follows: 1. Serum ELISA-IgG value was $0.37{\pm}0.134(Mean{\pm}S.D.)$ in vaginal trichomoniasis patients and $0.21{\pm}0.054$ in healthy controls(P<0.005), and the sensitivity and specificity of ELISA for serum IgG antibody were 70.0% and 96.7%, respectively. 2. Serum ELISA-IgM value was $0.33{\pm}0.177 (Mean{\pm}S.D.)$ in vaginal trichomonlasls patients and $0.11{\pm}0.051$ in healthy controls (p<0.005), and the sensitivity and specificity of ELISA for serum IsM antibody were 70.0% and 96.7%, respectively. 3, The ELISA-IgG values showed a significant correlation with ELISA-IgM values(r=0.77, p<0.005) , With above results, it is assumed that ELISA is a reliable method for the diagnosis of T vaginalis infection and simultaneous measurement of serum IgG and IgM with this technique is recommended.

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Evaluation of an Enzyme-Linked Imrnunosorbent Assay for the Detection of Aflatoxin $B_1$ from the Imported Cereals (수입곡물 중의 Alfatoxin $B_1$ 검출을 위한 효소면역측정법의 평가)

  • 손동화;박애란;이인원
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.355-361
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    • 1992
  • In order to evaluate an enzyme-linked immunosorbent assay(ELISA) for practical use in detecting aflatoxin $B_1(AFB_1)$ from cereals, we compared $AFB_1$ concentrations of samples contaminated artificially or naturally that were quantitated by the ELISA with those spiked or quantitated by HPLC. Cotton seed meals(19 items), rape seed meals(ll), soybean meals(9), and corns(3) imported from foreign countries were used as sample cereals. The standard curves of each cereal class showed that 1-100 ng/g of $AFB_1$ from cereals could be assayed by the ELISA. When artificially contaminated cereals were assayed by ELISA, the average recovery of AFB! from samples spiked to 3 ng/g and more was 138%(68-193%), although that spiked to 1 ng/g was somewhat high(268%). The average C.V. of recovery was 7.0%(0-22.2%). When naturally contaminated cereals were assayed, the concentrations of $AFB_1$ below 10 ng/g especially from rape seed meals quantitated by ELISA were much lower than those determined by HPLC. However, the concentrations of 10 ng/g and more from samples, except a few extraordinary samples. quantitated by ELISA were similar to those determined by HPLC, especially in case of cotton seed meals whose average recovery (ELISA/HPLC) was 153%. In conclusion, the ELISA was elucidated such as a practical tool to detect $AFB_1$ of 10 ng/g and more from cereals.

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Development of Enzyme-Linked Immunosorbent Assay for Glyphosate-Tolerant Soybeans (제초제내성 유전자재조합 콩의 검출을 위한 면역분석법 개발)

  • Kwak, Bo-Yeon;Ko, Seung-Hee;Park, Chun-Wuk;Son, Dae-Yeul;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.366-372
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    • 2003
  • Enzyme-linked immunosorbent assay (ELISA) for assaying the 5-enolpyruvyshikimate-3-phosphate synthase from Agribacterium sp. CP4 (CP4 EPSPS) in genetically modified soybeans was developed. Polyclonal and monoclonal antibodies (Pab, Mab) specific to the CP4 EPSPS were produced. When using the Pab, the detection limit of sandwich ELISA toward CP4 EPSPS (0.03 ${\mu}g/mL$) was better than that of competitive indirect ELISA(ciELISA) (1 ${\mu}g/mL$). It was found that 2 of 3 monoclonal antibodies, Mab1 and Mab2, recognized the same antigenic determinant on CP4 EPSPS, but Mab3 recognized different antigenic determinant when competitive ELISA was performed using the Mabs. On the other hand, when the sensitivity of sandwich ELISA using combination of Pab and/or Mabs was determined, the sandiwich ELISA using Mab2 as a capture antibody and Pab-HRP as a secondary antibody showed the lowest detection limit of CP4 EPSPS (0.02 ${\mu}g/mL$). The sandwich ELISA developed in this study could be applied to detect glyphosate-tolerant soybeans.

Diagnosis of swine sarcoptic mange(Sarcoptes scabiei var. suis) by ELISA (ELISA를 이용한 돼지 옴 (Sarcoptes scabiei var. suis) 감염증의 진단)

  • Jee, Cha-Ho;Lee, Sam-sun;Chang, Lai-hun
    • Korean Journal of Veterinary Research
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    • v.41 no.4
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    • pp.565-570
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    • 2001
  • The diagnosis of swine sarcoptic mange (Sarcoptes scabiei var. suis) was investigated by ELISA in order to replace current diagnostic methods such as skin scraping, scratching index, or lesion score of dermatitis. The current methods need many efforts and much times and cost much. They can not handle many samples simultaneously. Therefore, in this research we developed ELISA that can handle many samples at a time. The antigens of swine sarcoptic mite were isolated and examined by 12.5% SDS-PAGE and silver staining. The antigenicity of antigen was confirmed by Western blotting using the swine from the artificailly-infested swine with swine sarcoptic mite. The optical density (OD) values of the artificailly-infested positive sera and the naturally-infested positive sera of sows were measured and read in order to confirm the stability of antigens, the reproducibility and validity (sensitivity and specificity) of the manufactured ELISA of swine sarcoptic antigens. In above results, the developed ELISA would be possible to use the diagnostic tool of sarcoptic mange if OD values of piglets, fattening pigs and sows are interpreted reasonably and classified as mange-free and infested.

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Survey on the distributions of swine pleuropneumonia antibodies by ELISA in Kyongbuk western area (효소면역흡착시험을 이용한 경북서부지역의 돼지 흉막폐렴에 대한 항체분포조사)

  • 서희진;배성수;김대원;김봉환
    • Korean Journal of Veterinary Service
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    • v.23 no.3
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    • pp.289-299
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    • 2000
  • The study was performed to investigate the distributions of swine pleuropneumonia in Kyongbuk western area by the enzyme-linked immunosorbent assay (ELISA). Sera collected from 400 slaughtered pigs in 3 slaughter houses during the period from May 1999 to october 1999 were tested to detect antibodies against A pleuropneumonie serotype 2 and 5. The optimal dilution of CBE antigen, conjugate and serum for this ELISA were determined 1 : 400, 1 : 20,000, 1 100, respectively. The optimal dilution of OW antigen, conjugate and serum for this ELISA were determined 1 : In, 1 : 20,000, 1 : 200, respectively. Cut-off value in this ELISA was determined by mean absorbance (at 492 nm) of negative control sera added with the triple value of the standard deviation. Cut-off value in ELISA by CBE and OMP antigen were 1.134 and 1.217, respectively. By the ELISA, positive reaction rates to A pleuropneumoniae serotype 2 and 5 for CBE antigen were 38.8% and 18.8%, and for OMP antigen were 42.8% and 23.5% of the 400 samples, respectively.

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Studies on enzyme-linked immunosorbent assay(ELISA) for detection of antibody to Brucella abortus (효소면역법을 이용한 Brucella abortus 항체 검출에 관한 연구)

  • 심항섭;국정희;정봉수;고태오;조중현;박유순
    • Korean Journal of Veterinary Service
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    • v.21 no.2
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    • pp.107-115
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    • 1998
  • In order to establish a rapid, sensitive and specific diagnostic method for detection of antibody to Brucella abortus, a enzyme-linked immunosorbent assay(ELISA) was adapted. The diagnostic efficacy of the established ELISA was compared with that of the standard tube agglutination test for B abortus. 1. It was found that the optimal concentration of antigen for this ELISA was 5$\mu\textrm{g}$/ml, the optimal dilution of conjugate was 1 : 2000, and the optimal dilution of serum was 1 : 200, respectively. 2. Cut off value in this ELISA was 1,102 that was determined by mean absorbance(at 492nm) of tube agglutination test negative serum added with the triple value of the standared devation. 3. The relationship between the tube agglutination test and ELISA was showen high corresponding rate with sensitivity(96.3%) and specificity(98.1%). 4. The efficacy of the ELISA for detection of B abortus antibody was compared with tube agglutination test In brucellosis outbreak farm. The sensivity of ELSIA was higher than tube agglutination test.

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Comparison of a new ELISA with other serodiagnostic tests for bovine brucellosis (소 브루셀라병의 혈청학적 진단법 비교실험)

  • Hur, Jin;Kakoma, Ibulaimu;Jeong, Jae-Myong;Lee, Hyeon-Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.385-391
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    • 2007
  • A novel enzyme linked immunosorbent assay (ELISA) is described and compared with other established serologic tests for bovine brucellosis, namely the rose bengal test (RBT), the complement fixation test (CFT), and the tube agglutination test (TAT) approved and used in Korea. A total of 109 bovine serum samples were tested using all the 4 assays and analyzed as to specificity, sensitivity, reproducibility and predictive value. The ELISA showed 100% agreement with the CFT. The least agreement between ELISA was observed with the TAT. The agreement between the ELISA and RBT was not significantly different from that observed between the CFT and the ELISA. It is concluded that the new assay would be a good candidate for routine serologic survey for brucellosis in Korea. A protocol combining the ELISA and the CFT would increase the power for detection of serologically positive individuals and herds.