• 공업화학
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• 제22권5호
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• pp.522-525
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• 2011

본 연구에서는, enzyme-linked immunosorbent assay (ELISA)와 면역크로마토그래픽 기법을 결합하여 Staphylococcus aureus 검출을 위한 면역스트립을 제작하였다. 면역스트립은 4개의 서로 다른 기능을 가진 멤브레인을 이용하여 만들어졌다. 니트로셀룰로오스 멤브레인은 항체와의 결합력이 높기 때문에, 포획항체를 고정화하였고, 다공성 멤브레인들을 통하여 모세관 현상으로 인해 시료흐름을 유도하였다. 효소반응에 의해 생성된 발색신호는 디지털카메라와 자체 제작된 소프트웨어를 이용하여 정성, 정량분석 하였다. 최적의 분석조건 하에서 30 min 이내에 $2.7{\times}10^4{\sim}2.7{\times}10^7CFU/mL$ 범위의 S. aureus 농도를 측정할 수 있었다.

• 대한수의학회지
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• 제33권3호
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• pp.539-545
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• 1993

This study was carried out the determine the progesterone concentration for serum by enzyme-linked immunosorbent assay(ELISA) in bovine adult at estrous, pregnant, after patuation and male, female calves of 1 month old, respectively. The results obtained were summarized as follows ; 1. The assay has a sesitivity of $0.1ng/m{\ell}$. 2. Intra and inter-assay coefficient of variation were 4.5%, 6.1~9.4% when used for the determination of progesterone in samples of bovine serum. 3. The percentages of recovery for progesterone added were between 88.0 to 88.9%. 4. Progesterone concentration of adult bovine serum at estrus, pregnant and after 1 day of parturition were $0.37{\pm}0.16$, $7.1{\pm}1.0$, $0.13{\pm}0.02ng/m{\ell}$, respectively. 5. There was no differences in serum progesterone concentration of calves both male($0.16{\pm}0.03ng/m{\ell}$) and female($0.15{\pm}0.04ng/m{\ell}$) on 1 month old. From these results, progesterone determination by ELISA is very applicable to detect of early pregnancy diagnosis, factorial analysis of reproductive disorder, and also reproductive physiological functions such as veterinary therapeutic measures and monitoring of cyclicity.

• 한국식품위생안전성학회지
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• 제9권3호
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• pp.123-131
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• 1994

An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

• 대한수의학회지
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• 제57권1호
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• pp.31-36
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• 2017

Japanese encephalitis (JE) is an important zoonosis caused by the mosquito-transmitted JE virus (JEV), which is a causative agent of reproductive failure in pregnant sows. Detection of JEV antibodies in swine is performed by hemagglutination inhibition (HI), virus neutralization (VN), and the plaque reduction neutralization test (PRNT). The most stringent PRNT is the 90% endpoint PRNT ($PRNT_{90}$). These conventional assays are difficult to carry out in diagnostic laboratories with insufficient instruments or cell culture systems. An alternative assay that is easily conducted and time efficient is required. In this study, we improved the indirect enzyme-linked immunosorbent assay (I-ELISA) with clarified antigen for the detection of JEV antibodies. The I-ELISA results obtained from 175 swine serum samples were compared with HI, VN, and $PRNT_{90}$ results. The sensitivity of I-ELISA was 91.8%, 95.0%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. The specificity of I-ELISA was 92.2%, 94.7%, and 94.7% compared with HI, VN, and $PRNT_{90}$ results, respectively. Moreover, the I-ELISA results were significantly correlated with the HI (r = 0.93), VN (r = 0.95), and $PRNT_{90}$ (r = 0.92) results. These results suggest that the improved I-ELISA is useful for serosurveillance of JEV in swine.

• 대한수의학회지
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• 제37권3호
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• pp.613-618
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• 1997

곰팡이에 오염된 저질사료에서 T-2 독소의 존재확인 및 양을 측정하기 위한 Direct Competitive Enzyme Linked Immunosorbent Assay(ELISA)의 Kits를 개발하기 위하여 T-2HS, T-2HS-BSA, T-2HS-HRP 및 T-2 단크론 항체 동을 개발하고저 연구하여 좋은 결과를 얻었다. 분말 옥수수내에 인위적으로 혼합된 T-2 독소의 평균회수율은 83%이었으며, T-2 독소의 추출가능범위는 60ng에서 $6{\mu}g$이었다. 분말옥수수에서 인위적으로 혼합된 T-2 독소의 회수결과에 의하면 이 연구는 T-2 독소의 존재를 확인하기에 적합하고, T-2 독소의 양을 측정하기 위한각종 ELISA Kits 제조의 기틀이 마련되었다.

• 약학회지
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• 제38권3호
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• pp.351-357
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• 1994

The levels of maternal immunity enhancing factor(MIEF), which is an immunomodulatory protein identified from bovine colostrum, were determined by indirect competitive enzyme-linked immunosorbent assay(ELISA) for the colostrum and normal milk collected during the first two weeks of lactation. The mean concentration of MIEF in the colostrum of the first day of lactation was $109\;{\mu}g/ml$, and fell from the third day of lactation to $3{\sim}4\;{\mu}g/ml$. The molecular weight of the purified MIEF determined by reducing SDS-PAGE and TSK G2000SW column chromatography was 22,000 and 24,000 daltons, respectively, showing that MIEF is a monomeric peptide in its native form. To examine the capacity of MIEF to induce differentiation of B Lymphocytes, human tonsillar Iymphocytes were cultured in the presence of different concentrations of MIEF, and then antibody secreting cells were enumerated by enzyme-linked immunospot(ELISPOT) assay. When added to cultures of human tonsillar Lymphocytes, MIEF induced differentiation of resting B Iymphocyte to antibody secreting plasma cells as efficiently as LPS.

• Journal of Microbiology and Biotechnology
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• 제18권11호
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• pp.1858-1861
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• 2008

A PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed for the rapid and sensitive detection of L. monocytogenes. PCR primers generating a 132-bp amplicon and a capture probe able to hybridize to the PCR amplicon were designed based on the L. monocytogenes-specific hly gene encoding listeriolysin. The detection limit of PCR-ELISA for L. monocytogenes was determined to be as low as 10 cells per PCR reaction, and this level of detection was achieved within 5 h. These results indicate that the PCR-ELISA provides a valuable tool for the rapid and sensitive detection of L. monocytogenes for the ready-to-eat food industry.

• 한국식품과학회지
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• 제32권3호
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• pp.538-543
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• 2000

가열 처리된 우육, 돈육, 계육, 양육과 염소육과의 종판별을 위한 효소면역측정법(ELISA)을 확립하기 위하여 염소육을 $98^{\circ}C$에서 15분간 열처리해서 추출액을 분리하였다. 열에 안정한 주요 단백질(TS)은 분자량이 36과 38kDa이었고 두 분자량대의 단백질의 pI값은 4.5로 확인되었으며 면역원으로 사용하기 위해 이온교환 (DEAE-Sephadex A-50)과 겔 여과(Sephadex G-75) 크로마토그래피로 정제하였다. 분리, 정제된 TS단백질을 토끼에 면역하여 염소육에 대해 특이성을 갖는 항체를 생산하고 항TS 항체를 이용하여 간접경합 ELISA를 확립하였다. 항TS 항체는 가열 처리된 계육, 우육, 돈육, 양육과는 반응성이 전혀 없었고 동일하게 처리된 염소육과 지표단백질인 TS에만 반응성을 보였다. 따라서, 본 연구에서 확립한 간접경합 효소면역측정법은 가열 처리된 육류 중 염소육 판별에 웅용 가능함을 시사하였다.

• 한국어병학회지
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• 제9권2호
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• pp.169-176
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• 1996

연어과 어류의 이상유영 원인 바이러스 RVS의 ELISA법에 의한 신속 진단 방법을 개발하였다. 주화세포를 이용한 바이러스 배양액 및 감염 무지개송어의 뇌조직 마쇄액을 사용하여 실험하였다. 바이러스 배양액을 이용한 ELISA법의 검출 감도 조사에서 최소 바이러스 감염가 검출 한계치는 $10^{2.6}$ $TCID_{50}/100{\mu}l$ 이었다. 또한, 인공감염어의 뇌조직 마쇄액 내의 RVS 항원도 검출 되었다. 본 방법은 현장에서의 RVS 감염어 조사에 효과적으로 사용되어질 수 있는 방법으로 생각 되어진다.

• Environmental Analysis Health and Toxicology
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• 제28권
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• pp.16.1-16.8
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• 2013

Objectives Fish vitellogenin (VTG) is produced in the female liver during oogenesis through the estradiol cycle and produced in the male liver by endocrine disrupting chemicals (EDCs) such as alkylphenols. In this study, we propose that the VTG concentration in the pale chub could be detected using monoclonal antibodies and polyclonal antibodies against vitellin (Vn) in a VTG enzyme-linked immunosorbent assay (ELISA) system. Methods Monoclonal antibodies and polyclonal antibodies were produced using the Vn extracted from the matured ovum of the ovary. The VTG was extracted from the plasma of the male pale chub. The Vn and VTG were confirmed by measuring the molecular weight of their proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the specificity of the antibodies was checked through western blotting methods. The assay system was validated with respect to optimal assay concentrations, specificity, recovery, and intra- and inter-assay variations. Results The Vn consisted of two protein bands with apparent molecular weights of 64 and 37 kDa. The SDS-PAGE indicated protein weights of 146 and 77 kDa in the VTG. The assay range was 15.6 ng/mL to 2,000 ng/mL, and the value of the intra- and inter-assay variations were within 10.0% and 14.7%, respectively. The recovery rate was $99.5{\pm}5.5%$. Conclusions A sandwich ELISA was developed that could be used to qualify the VTG of pale chub in screening for EDCs. Pale chub is an ideal species for observing estrogen activity in the environment because of its extensive habitat and extensive food chain. The ELISA developed here would be more favorable than those for other species for determining the effect of long-term food chain accumulation of EDCs in aquatic environments.