• Title/Summary/Keyword: ELISA

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Evaluation of Two ELISA and Two Indirect Hemagglutination Tests for Serodiagnosis of Pulmonary Hydatid Disease

  • Eris, Fatma Nur;Akisu, Ciler;Aksoy, Umit
    • Parasites, Hosts and Diseases
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    • v.47 no.4
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    • pp.427-429
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    • 2009
  • To establish a definite diagnosis for pulmonary hydatid disease, combination of radiology and serology is useful. In this study, 19 preoperative sera from patients with surgically confirmed pulmonary hydatidosis, 40 sera from patients with other parasitosis and pulmonary diseases, and 20 sera from healthy donors were evaluated using 4 different serological tests, i.e., the commercial ELISA (ELISA-kit) test, the ELISA (ELISA-lab) test prepared in our laboratory, the commercial indirect hemagglutination assay kit (IHA-kit) test, and the IHA test using sensitized sheep red blood cells with tannic acid (IHA-TA). The ELISA-kit was the most sensitive (84.2%) and the most specific test (100.0%). The ELISA-kit also demonstrated the highest positive (100.0%) and negative (95.2%) predictive values. The sensitivity of the ELISA-lab test, that we prepared, was found to be 73.6%, whereas the IHA-kit test and the IHA-TA test were found to be 73.6% and 68.4%, respectively. The specificity of these tests was 96.6%, 98.3%, and 83.3%, respectively. When all 4 tests were assessed together, it was found that the sensitivity had risen to 94.7%. When the ELISA-kit was assessed with the IHA-kit and IHA-TA together, it was found that the sensitivity was 89.5% and 84.2%, respectively. Likewise, the combination of the ELISA-lab and IHA-kit or IHA-TA allowed us to achieve a sensitivity of 84.2% in cases of pulmonary echinococcosis. In conclusion, the diagnosis would be imminent if least 2 tests were applied together.

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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A Comparative Study of Serologic Methods for Detection of Mumps Antibody in Korean Children (한국 소아의 Mumps 바이러스 항체보유에 관한 혈청학적 진단방법의 비교)

  • Park, Hae-Kyung
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.473-480
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    • 1986
  • Mumps is an extremly common infectious disease affecting predominantly young children hut it is not a severe disease in terms of mortality. One hundred and two sera from infants of 3 different groups which are vaccinated, unvaccinated and unknown were detected to mumps antibody. The tests used were Complement Fixation(CF) test, Single Radial Hemolysis(SRH) test, Hemagglutination Inhibition(HI) test, Enzyme Linked Immunosorbent Immunoglobulin G(ELISA IgG) test, Enzyme Linked Immunosorbent Immunoglobulin M(ELISA IgM) test. 1. The rate of positivity for mumps antibody in 102 sera wera 89.16%(74/83) by Hl test, 68.83%(53/77) by ELISA IgG test, 64.58%(62/96) by SRH test, 63.24%(43/68) by ELISA IgM test and 50.00%(49/98) by CF test. 2. The rate of positivity by 5 tests for 55 sera turned out to be very similar with above results respectively. 3. The correlation coefficients(r) between ELISA IgG test ant H1 test, ELISA IgG test and ELISA IgM test were 0.34(P<0.0l) and 0.31(P<0.02), respectively. 4. The percentage of apparently natural infection of mumps seemed to be 65.15%(43/66) in infants. 5. Seroconversion rate of mumps by vaccination were 90.91%(10/11). 6. Among the 53 infants who were tested with ELISA IgG 15 were below 15 months age of(28.30%) and this percentage may be taken as a suggestion that mumps vaccination should be given earlier than present practice. 7. ELISA IgG test was found very sensitive and recommendable method for large scale screening for the presence of antibody to mumps.

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Changes of maternal antibodies in chicks vaccinated to breeder against infectious bronchitis, infectious Bursal disease, and Newcastle disease virus (모계의 전염성기관지염, 전염성 F낭병 및 뉴캣슬병 백신투여에 따른 모체이행 항체의 변동)

  • 고원석;김태중;이정원;서이원;송희종;오언평
    • Korean Journal of Veterinary Service
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    • v.21 no.2
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    • pp.133-139
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    • 1998
  • Serum samples collected from 30 breeders and their progeny 30 chicks. The antibodies against infectious bronchitis(IB), infectious bursal disease (IBD) and Newcastle disease(ND) viruses were detected by ELISA using commercial ELISA kit. The breeders were vaccinated against IB, IBD and ND viruses according to general vaccination program. Geometric mean titers(GMT) of ELISA were monitored from 1-day old to 17-day old chicks and compared with breeder chickens. The GMT of ELISA to IB, IBD and ND were declined half level of the breeder antibody titer at 6-, 8- and 7-day old. And, the GMT of ELISA to IB, IBD and ND were declined than that of protective titer at 6-, 1-, and 4-day old. Thereafter, the GMT of ELISA was declined and disappeared according to ages of chicks. Taken together, this study led to conclusion that time-course of maternal antibody titers of chicks from vaccinated breeders, and this is very important data for vaccination to chicks.

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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
    • Parasites, Hosts and Diseases
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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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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.

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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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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