• 제목/요약/키워드: Ci-ELISA

검색결과 77건 처리시간 0.019초

판토텐산의 분석을 위한 효소면역측정법 (An Enzyme-Linked Immunosorbent Assay for Detection of Pantothenic Acid)

  • 손동화;박윤식;배근원
    • 한국식품과학회지
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    • 제32권5호
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    • pp.1009-1014
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    • 2000
  • PA를 분석하기 위하여 효소면역측정법을 개발하고자 하였다. Bc방법과 Po방법으로 BSA에 PA를 conjugation하여 각각의 PA-BSA conjugate(PA-BSA[Bc]와 PA-BSA[Po])를 제조하였으며, 이를 토끼에 면역하여 항PA-BSA 항체를 얻었다. 항PA-BSA[Po] 항체를 사용한 ciELISA의 결과에서 경합반응이 제대로 일어나지 않았기 때문에, 식품 속에 있는 PA를 검출하기 위해서 PA-BSA[Po]을 코팅한 후 항PA-BSA[Bc] 항체를 사용하였다. 이 결과에서 PA의 검출한계가 1 ppm인 것을 확인할 수 있었으며, 교차반응을 통해 PA 유도체들에 대해서도 항PA-BSA[Bc] 항체가 PA에 대해 특이성이 매우 강하였다. 또한, MBA의 결과에서는 그 검출한계가 10ppb인 것을 확인할 수 있었다. 분석시료인 계란(109%), 상추(64%), 소간(344%)의 식품시료에 대한 실험에서 상추를 제외하고는 ciELISA는 MBA의 결과와 비교해 볼 때 양호한 결과를 얻을 수 있었다. 그러므로, ciELISA는 MBA보다 분석시간, 교차반응 등의 면에서 장점이 있어 식품 중 PA의 검출에 효과적으로 활용할 수 있을 것이다.

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Immunological Assay to Detect Irradiated Beef

  • Lee, Ju-Woon;Yook, Hong-Sun;Lee, Hyun-Ja;Kim, Jung-Ok;Byun, Myung-Woo
    • Preventive Nutrition and Food Science
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    • 제6권2호
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    • pp.91-95
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    • 2001
  • Competitive indirect enzyme linked immunosorbent assay (Ci-ELISA) was used to obtain the preliminary data for the detection of irradiated beef. Ci-ELISA was individually formatted with polyclonal antibodies produced from 2 kinds of bovine proteins, myosin and bovine serum albumin (BSA). Beef round, loin and tender loin were vacuum-packaged and subdivided into 3 groups of 1) irradiation; 2) irradiation and chilled at 4$^{\circ}C$ for 7 day; 3) irradiation and frozen at 2$0^{\circ}C$ for 2 months to observe the changes under different storage and/or distribution conditions. Irradiation was performed at 3, 5 and 7 kGy. Protein solutions prepared from the sample were tested by formatted Ci-ELISA. Detected concentrations of myosin and BSA decreased with the increased irradiation dose in all samples with different reduction rates. Myosin was more susceptible to freezing than BSA. Samples irradiated at 5 kGy or above could be differentiated from non-irradiated ones by Ci-ELISA. These results indicate that immunological assay can be used as a detection method for irradiated beef.

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가공식품 중 메밀 검출을 위한 경합 ELISA의 개발 (Development of Competitive Indirect ELISA for the Detection of Buckwheat in Processed Foods)

  • 백수연;도정룡;손동화
    • 한국식품과학회지
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    • 제46권3호
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    • pp.269-275
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    • 2014
  • 메밀로부터 분리한 메밀단백질(BWP)을 토끼에게 면역하여 생산한 특이항체를 이용하여 가공식품 중 메밀의 검출을 위한 간접경합 효소면역측정법(ciELISA)을 확립하였다. 이때, BWP의 검출범위는 $0.05-100{\mu}g/mL$이었다. 특이항체 교차반응 결과, BWP, 메밀가루, 통메밀에 대한 반응성은 각각 100, 17.9, 11.8%를 나타내었으며 그 외 곡물 14종에 대한 반응성은 거의 나타나지 않아 메밀에 대한 특이성이 매우 높았다. 메밀가루는 $60-90^{\circ}C$에서 반응성이 평균 83.0%를 나타내었으며 $100^{\circ}C$에서는 44.5%로 반응성이 감소하였다. 메밀가루에 대한 spike test에서 메밀 생면, 삶은 면, 묵, 메밀곡물가루에서 평균 분석회수율이 각각 99.1, 98.6, 81.1, 104%로 매우 높게 나타났다. ciELISA에 의하여 22점의 시판시료 중 메밀의 함유량을 조사한 결과, 메밀 함유 표시된 12점에서 평균 분석회수율은 31.6%로 나타났으나 시료 중 메밀의 정성적 검출은 100%로 매우 우수하였다. 본 연구에서 개발한 ciELISA는 가공식품에 함유된 메밀을 검출하는데 매우 효과적으로 활용할 수 있을 것이다.

돼지생식기호흡기증후군(PRRS) 바이러스 감염 항체 검출 ELISA 상용 키트의 정확도 비교 (Comparison of Two Commercial Antibody Enzyme-Linked Immunosorbent Assays for Detection of Porcine Reproductive Respiratory Syndrome Virus Infection)

  • 박선일;이승환;박경애
    • 한국임상수의학회지
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    • 제33권2호
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    • pp.102-106
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    • 2016
  • More than 20 years after the first report of porcine reproductive and respiratory syndrome virus (PRRSV) in Korea, the disease is still having major impact on domestic pig health and relevant industries. Although ELISA tests are commonly used by veterinarians to guide herd management, data on diagnostic performance of the test in field settings are very limited. The objective of this study was to evaluate two commercially available PRRSV ELISA (IDEXX PRRS X3 ELISA and Bionote PRRSV ELISA 4.0) to detect antibodies against PRRSV on serum samples. To this end, a total of 1,108 sera were recruited from 35 swine farms located in Gyeonggi province and tested at the Gyeonggi Province Veterinary Service Center. All tests were performed according to the manufacturer's instructions, by laboratory technicians who routinely perform PRRS testing on blood samples. Samples were collected from two sources of swine populations with different PRRS prevalence; 60 samples (5.4%) were originated from breeding farms and the remaining 1,048 samples (94.6%) were from farrow-to-finish farms. We applied Bayesian latent class model (LCM) for two-tests in the two-population when the accuracy of the gold standard is not available. The model estimated that Bionote ELISA was a bit more specific but slightly less sensitive. The estimated sensitivity and specificity of the IDEXX ELISA were 99.8% (95% CI 98.1-100%) and 86.4% (95% CI 81.4-96.5%), respectively. Sensitivity, specificity, positive predictive value and negative predictive value for Bionote kit were 98.7% (95% CI 92.8-100%), 89.8% (95% CI 86.2-93.1%), 93.8% (95% CI 91.5-96.0%), and 97.8% (95% CI 87.1-100%), respectively. Based on the Bayesian 95% credible intervals, the sensitivity and specificity of the two ELISAs were not significantly different each other when assuming that two kits were imperfect, indicating that two kits performed equally well in terms of sensitivity and specificity in our filed setting.

Enzyme-linked Immunosorbent Assay for the Detection of Hen's Egg Proteins in Processed Foods

  • Shon, Dong-Hwa;Kim, Hyun-Jung;Kim, Soo-Ho;Kwak, Bo-Yeon
    • 한국축산식품학회지
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    • 제30권1호
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    • pp.36-41
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    • 2010
  • The Hen's egg is widely used in many processed foods as an ingredient and is one of the most prevalent food allergens in children. To detect egg proteins in processed foods, we developed a competitive indirect enzyme-linked immunosorbent assay (ciELISA) using an anti-ovomucoid (OM) antibody, which was produced by immunization of rabbits with OM, the most heat-stable component of the egg proteins. The detection limit of this quantitative assay system was 30 ng/mL. Cross-reactivity of the anti-OM antibody toward OM, ovalbumin, skim milk, casein, whey protein isolate, and isolated soy protein was 100, 0.4, 0.2, 0.04, 0, and 0%, respectively. In the spike test of egg white powder in milk replacer, commercial sausage, and in-house sausage, the assay recoveries ($mean{\pm}SD$) were $129{\pm}13.7%$, $73.9{\pm}12.5%$, and $65.5{\pm}13.6%$, respectively. When egg white in a commercial crab meat analog and sausage was determined by ciELISA, the assay recovery was found to be 108% and 127%, respectively. The combined results of this study indicate that this novel ciELISA for OM detection could be applied for the quantification of hen's egg proteins in processed foods.

Lipopolysaccharide Yields from Rhodobacter capasulatus with indirect ELISA

  • Yoo, Tae-Eun;Lee, Hyun-Soon
    • Journal of Microbiology
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    • 제34권3호
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    • pp.255-262
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    • 1996
  • The lipopolysaccharide (LPS) yields were measured in Rhodobacter capsulatus under several conditions by the ELISA method. The purification of LPS was done by affinity chromatography of IgG coupled CNBr-activated sepharose-4B instead of ultra-centrifugation. The purity of the LPS didn't show much difference between affinity chromatography and ultra-centrifugation method, but affinity chromatography method required much fewer organisms and was more convenient. LPS yield was measured in ng units by the ELISA method. Mannitol was a better single carbon source than other sugars, but mixing two carbon sources resulted in greater LPS yields than any sugar alone. LPS yield was directly proportional to $NH_ 4CI$ concentration, with optimum yields at 0.05% nitrogen. In contrest to LPS yields, which decreased at 0.005% nitrogen concentration total protein was increased 16 times. Calcium influenced LPS yields. At 0.7 mM $CaCI_ 2$, the LPS yield was 16.5 $\mu$g/mg DW, five times the yield without calcium.

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효소면역측정법을 위한 다클론 항대두단백 항체의 생산 및 특성비교 (Property Comparison of Polyclonal Anti-Soy Protein Antibodies Produced for ELISA)

  • 손동화;김현정;윤숭섭
    • 한국식품과학회지
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    • 제32권6호
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    • pp.1221-1226
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    • 2000
  • 대두단백질의 분석을 위한 효소면역측정법을 개발하고자 특이항체를 생산하고 그 항체의 특성을 비교하였다. 분리대두단백(ISP), ISP를 SDS와 urea의 첨가하에 열처리한 것(ISP(SU)), 11S 글로불린의 acidic subunit(AS)를 각기 면역하여 다클론 항체를 생산하였다. 간접 경합 효소면역측정법(ciELISA)을 실시하여 처리를 달리한 대두단백질에 대한 이들 항체의 반응성을 조사하였여 $IC_{50}$으로 나타내었다. 항AS 항체의 경우 $IC_{50}$값이 ISP, ISP(SU), ISP를 2-ME로 처리한 ISP(ME), crude 11S에 대하여 각각 20, 2, 2.5, $200\;{\mu}g/mL$로 나타났다. 또한, 항ISP(SU) 항체의 경우 같은 항원에 대해서 100, 5, 4, $220\;{\mu}g/mL$였으며 항ISP 항체의 경우는 각각 20, 30, 36, $1000\;{\mu}g/mL$로 나타났다. 이로서 생산된 3가지 항체 중에서 항AS 항체가 대두단백질에 대한 반응성이 가장 우수하여 ELISA분석법 개발에 적합하였다.

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