• 제목/요약/키워드: enzyme-linked immunosorbent assay(ELISA)

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Chitooligosaccharides in Korean Commercial Salt-Fermented Shrimps, Determined by Enzyme-Linked Immunosorbent Assay

  • Shim, Youn-Young;Shon, Dong-Hwa;Chee, Kew-Mahn
    • Journal of Microbiology and Biotechnology
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    • 제14권4호
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    • pp.877-880
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    • 2004
  • In this study, we determined the content of chitooligosaccharides (COS) in Korean commercial salt-fermented shrimps by competitive direct enzyme-linked immunosorbent assays (cdELISAs), using anti-COS mixture (COSM) antibody and COSM horseradish peroxidase (HRP) conjugate. When COS6 was spiked into salt-fermented shrimps at the level of $10-300\mu{g/g,}$ the average recovery was $120\pm19%$ ($mean\pmS.D.$). The COS contents of the 92 samples of Korean commercial salt-fermented shrimps collected during February 2000 and August 2002 were $36.3\pm20.7\mug$ COS6 equivalent/g (expressed as "$\mug/g$" hereafter). Among the samples, the COS contents of yuk-jeot ( $40.3 \pm 22.5 \mug/g, n=27$) and buksaewoojeot ($40.2 \pm 21.6 \mug/g, n=5$) were higher than the others. The COS contents of salt-fermented shrimps produced at Gwangcheon ($47.1 \pm 20.7 \mug/g, n=18$) and Gomso ($44.1 \pm 21.8 \mug/g, n=6$) areas were higher than those produced at the other areas. This is the first report to determine COS of salt-fermented shrimps by cdELISA.

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

  • 서희진;배성수;김대원;김봉환
    • 한국동물위생학회지
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    • 제23권3호
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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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ELISA와 RT-PCR에 의한 국내재배난에서 심비디움 모자이크 바이러스와 오돈토글로섬 윤문 바이러스이 검정 (Detection of Cymbidium Mosaic Virus and Odontoglosum Ringspot Virus by ELISA and RT-PCR from Cultivated Orchids in Korea)

  • 박원목;심걸보;김수중;류기현
    • 한국식물병리학회지
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    • 제14권2호
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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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살균제 Iprovalicarb 잔류물의 신속한 검출을 위한 효소면역분석법 (Enzyme Immunoassay for Rapid Detection of the Fungicide Iprovalicarb Residues)

  • 조한근;경기성;이은영
    • Journal of Biosystems Engineering
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    • 제31권6호
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    • pp.535-540
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    • 2006
  • For a biosensor development, an enzyme-linked immunosorbent assay (ELISA) of the fungicide iprovalicarb was developed by minimizing the processing time. The time for whole incubation process was reduced from 135 minutes to 15 minutes. The concentration of antibody was varied to improve sensitivity. The total processing time was reduced from 2.5 hours to 20 minutes, the final sensitivity ($IC_{50}$ value) of 7.93 ng/mL and the lowest detection limit of 0.045 ng/mL were obtained. This ELISA was applied to potatoes and onions, and the recoveries were in the range of 98.85 $\sim$ 101.20% and 87.97 $\sim$ 102.70%, respectively. Accordingly, this method can be used as basis for a biosensor for rapid monitoring of iprovalicarb residues in crops.

살충제 이미다크로프리드 잔류물의 실시간 측정용 효소면역분석법 (Enzyme Immunoassay for On-line Sensing of the Insecticide Imidaclopird Residues)

  • 송석진;조한근
    • Journal of Biosystems Engineering
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    • 제28권6호
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    • pp.505-510
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    • 2003
  • In Korea, due to its broad efficacy as a systemic insecticide, imidacloprid has been widely used in rice paddies to control sucking insects, soil insects, and some chewing insects and in apple orchards to control various insects pests. To quantify the imidacloprid residue concentrations, samples are assayed in vitro using enzyme-linked immunosorbent assays(ELISA). These assays generally require several hours to perform. As a biosensor, a competitive imidacloprid ELISA was modified to measure insecticide concentrations. It was found that a total assay time of 15 min(10-min antibody-antigen binding, and 5-min substrate development) is sufficient for monitoring imidacloprid concentrations. Further work is needed to improve the sensitivity of the measurement protocol.

Comparison of an Immunochromatographic Assay Kit with DAS-ELISA for Large-Scale Diagnosis and Molecular Discrimination of Satsuma Dwarf Virus Collected from Citrus Orchards

  • Kato, Mitsuhiro;Tomimura, Kenta;Ishii, Kanako
    • The Plant Pathology Journal
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    • 제36권5호
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    • pp.509-514
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    • 2020
  • Satsuma dwarf virus (SDV) seriously damages citrus production by reducing the quality and yield of fruit. To avoid contamination with SDV, mother trees are checked to be SDV-free in advance of nursery tree distribution. In this study, we compared an immunochromatographic assay (ICA) kit with double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for large-scale diagnosis of SDV in orchardgrown trees in Shizuoka Prefecture, Japan. The two methods gave conflicting results for 11 of 1,705 samples, all of which were negative by DAS-ELISA but positive by ICA. The samples scored as positive by either DAS-ELISA or ICA were analyzed by reverse transcription polymerase chain reaction and all were confirmed to be positive. These results validate the use of ICA as a screening method for large-scale diagnosis. Strain discrimination revealed that 16 of 22 isolates belonged to SDV, while citrus mosaic virus (CiMV) infection only and co-infection (SDV and CiMV) were in a minority.

추출물의 희석에 의한 옥수수 중 Fumonisin의 효소면역측정법 (An Enzyme-Linked Immunosorbent Assay for Fumonisins in Corn without Cleanup Procedure)

  • 손동화;김영목
    • 한국식품과학회지
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    • 제28권5호
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    • pp.953-958
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    • 1996
  • 발암성 진균 독소의 하나인 fumonisin을 신속, 간편하게 분석할 수 있는 효소면역측정법 (ELISA)을 개발하고자 하였다. 먼저 Fumonisin $B_1\;(FB_{1})$에 특이적인 항체를 생산하기 위하여 $FB_1$을 keyhole lympet hemocyanin (KLH)에 공유결합시키고 이를 Freund's adjuvant와 함께 토끼에 3차례 피하주사하여 면역하였다. 높은 항체가와 가장 양호한 경합반응을 보인 항혈청으로부터 항체를 정제하였으며, 이 항체의 fumonisin 유사독소에 대한 교차반응은 fumonisin $B_1,\;B_2$$B_3$에 대하여 각각 100%, 69% 및 166%이었다. $FB_1$의 검출을 위하여 확립한 직접법 경합ELISA (cdELISA)로 옥수수에 인위적으로 오염시킨 $FB_1$을 정제없이 ELISA로 분석하는 경우, 75% methanol로 시료의 추출 후 완충액으로 1/100 희석하였을 때 양호한 회수율을 보였다. 이 조건하에서 행한 ELISA의 FB₁분석회수율은 $1-30\;{\mu}g/g$의 오염농도범위에서 평균 67%이었으며, 농도별 회수율의 분산은 3.4%로 분석치가 매우 안정하였다. 본 연구에서 개발한 fumonisin 분석용 ELISA system은 수입 및 국내산 농산물로부터 이 독소의 정제없이 손쉽게 다량의 시료를 일차로 분석하는 데 유용할 것으로 판단된다.

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희석에 의한 우유 중 $Aflatoxin\;M_1$의 효소면역측정법 (An Enzyme-Linked Immunosorbent Assay for $Aflatoxin\;M_1$ in Cow's Milk without a Cleanup Procedure)

  • 손동화;임선희;이인원
    • 한국식품과학회지
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    • 제28권6호
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    • pp.1184-1187
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    • 1996
  • 우유 중에 존재할 수 있는 발암성 진균독소의 하나인 $aflatoxin{\;}M_1{\;}(AFM_1)$을 신속, 간편하게 분석할 수 있는 효소면역측정법(ELISA)을 개발하고자 하였다. 소혈청알부민에 공유결합한 $AFM_1\;(AFM_1-BSA)$을 토끼에 면역하여 항체를 생산하고 정제하였다. 이 항체의 유사독소와의 교차반응율은 29.9%이하의 비교적 낮은 교차율을 나타냈다. $AFM_1$의 검출을 위하여 확립한 직접경합ELISA (cdELISA)로 우유에 인위적으로 오염시킨 $AFM_1$을 정제과정없이 ELISA로 분석하는 경우, 우유를 PBS로 40%되게(2:3) 희석하였을 때 양호한 회수율을 보였다. 이 조건하에서 행한 ELISA의 $AFM_1$분석 회수율은 0.3-3.0 ng/ml의 오염농도 범위에서 농도별 회수율로 평균 113%, 그 분산은 8.2%였다. 본 연구에서 개발한 ELISA system은 우유 중의 0.5 ppb이상의 $AFM_1$을 정제과정없이 손쉽게 분석하는데 유용할 것으로 판단된다.

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Detection of PCB77 by Indirect Competitive Enzyme-linked Immunosorbent Assay in Sea Sediment Samples

  • Chen, Han-Yu;Zhuang, Hui-Sheng;Yang, Guang-Xin
    • Bulletin of the Korean Chemical Society
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    • 제34권3호
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    • pp.922-926
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    • 2013
  • 3,3',4,4'-Tetrachlorobiphenyl (IUPAC PCB77) is one of seven indicative polychlorinated biphenyls (PCBs) in the surface sediments. The current study presents a novel polyclonal antibody for the determination of the PCB77 using indirect competitive enzyme-linked immunosorbent assay. Under optimum conditions, PCB77 was determined within the concentration range of 0.01-100 ${\mu}g\;L^{-1}$, with a detection limit of 0.057 ${\mu}g\;L^{-1}$. The assays were tested for their cross-reactivity profiles using 3 selected congeners and 4 Aroclor products. The assays were highly specific for coplanar PCB congeners, but less specific for Aroclor1248. The spiked recoveries from five sediment samples were 86%-114% for PCB77 from ELISA, which were satisfactory. The current study demonstrated that the developed antiserum and immunoassay procedure can be used to detect PCB77 in environmental samples. The results of the sediment analysis were confirmed by conventional GC/ECD.

Seroprevalence of Babesia gibsoni in Companion Dogs in Korea by Enzyme Linked Immunosorbent Assay using Recombinant BgTRAP Antigen

  • Kim, Mi-jung;Chung, Tae-ho;Na, Yun-hee;Choi, Ul-soo
    • 한국임상수의학회지
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    • 제34권3호
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    • pp.185-188
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    • 2017
  • This study assessed the seroprevalence of Babesia gibsoni in companion dogs in Korea by enzyme linked immunosorbent assay using recombinant BgTRAP antigen. Dogs were randomly selected from those admitted for various reasons to local private veterinary hospitals and the Animal Medical Center of Chonbuk National University. With the owners' permission, extra blood was drawn from each dog for serological assays. Of the 188 selected dogs, seven (3.72%) were positive for B. gibsoni, including six of 167 (3.59%) indoor and one of 12 (8.33%) outdoor dogs. Of the seven dogs positive for B. gibsoni, four were aged > 10 years, two were < 1 year, and one was between 1 and 10 years; and two were Yorkshires and one each was Shih-tzu, Maltese, Pekinese, beagle and mixed. Concurrent diseases or chief complaints were anemia in two dogs, both of which had a history of confirmed babesiosis by polymerase chain reaction, and non-anemic diseases in five. Geographically, four dogs were from Jeonbuk/Jeonju, and one each from Seoul, Gyounggi-do, and Jeonnam/Gwangju. To our knowledge, this is the first report of companion dogs in Korea being seropositive for B. gibsoni. Serologic screening of subclinical or carrier dogs can detect this potentially dangerous disease and assess its epidemiology.