• Asian-Australasian Journal of Animal Sciences
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• 제9권3호
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• pp.299-302
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• 1996

Milk samples(n = 78) were taken 19d, 20d, 21d, 22d after artificial insemination(AI) for early pregnancy diagnosis by using the Enzyme immunoassay(EIA) kit. The progesterone ($P_4$) concentration in the whole milk was measured on the same day of pregnancy diagnosis. Rectal palpation(RP) was accomplished between 60d and 70d after AI to estimate the ovary condition and pregnancy status. Milk progesterone concentrations measured by Radio-immunoassay(RIA) method, in the pregnant cows at 17d, 19d, 21d after insemination were $17.10{\pm}0.91$, $17.60{\pm}0.46$, and $18.43{\pm}0.79nmol/l$, whereas those in the not-pregnant cows were $6.57{\pm}1.03$, $2.63{\pm}0.29$, and $0.67{\pm}0.08nmol/l$, respectively. When the progesterone concentration was less than 7 nmol/l, the color of the EIA kit was lighter and when the progesterone concentration was ${\geq}16nmol/l$, the color of the EIA kit was darker compared to the standard color. The detection rates of error by judging the color differences were 5.1% and 20.7%, respectively. In the early pregnancy diagnosis by the EIA kit and RIA method, the accuracy rates in the pregnancy of cows were 82% and 87%, and those in not-pregnant cows were 86% and 91%, respectively. For ovarian status estimated by the RIA method and certified by RP, the accuracy rates of the ovarian atrophy, follicular cyst and luteal cyst were 80, 91 and 83% and the progesterone concentrations were 2.51, 2.03, and 26.7 nmol/l, respectively.

• Parasites, Hosts and Diseases
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• 제21권2호
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• pp.265-269
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• 1983

간흡충증 환자 혈청에서 Thin layer immunoassay에 의하여 항체를 정량적으로 측정하여 감수성과 특이성을 알아보고, 효소표식면역 법(ELISA)에 의한 성적과 비교하였다. 냉동 건조한 간흡충 성충의 생리식염수 추출물을 항원으로 사용하였고, TIA는 Elwing등(1976)의 방법으로, 효소표식면 역법은 Veiler등(1974)의 방법에 의해 실시하였다. 1. 간홉충중 환자 60명의 혈청에서 TIA zone 크기의 평균은 4.14mm였고, 3mm 이상을 양성으로 판정하였을 때 감수성은 100%, 특이성은 61.1%였다. 2. 효소표식면역 법에 의하면 ELISA치 0.8이상을 양성으로 판정하였을 때 감수성은 88.3%, 특이성은 87.3%였다. 3. 혈청내 IgG 농도는 TIA zone 크기와 상관이 없었으나 좌소표식면역법에 의한 ELISA치와는 상관이 있어 그 계수는 0.69였다. 4. TIA zone 크기는 효소표식면역 법에 의한 ELISA치와 그 분포에 있어 상관이 없었다.

• Asian-Australasian Journal of Animal Sciences
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• 제10권6호
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• pp.603-608
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• 1997

To improve animal production, a simple and accurate pregnancy diagnosis plays a very important role. Therefore, the purpose of this study was to develop an on-farm blood progesterone enzyme immunoassay (EIA) system for monitoring the early pregnancy in sows. Star tubes coated with mouse monoclonal anti-progesterone antibody were used for this proposed EIA system which was tested in field trials. The results could be obtained within 30 minutes either by spectrophotometry or the naked eye. Heparinized fresh blood samples collected from the ear vein of sows 17-22 days after breeding (day 0) were tested qualitatively to diagnose sows as pregnant or non-pregnant with high ( > 3 ng/ml) or low ($${{\leq_-}}3ng/ml$$) progesterone in the blood. To provided a double check data, plasma progesterone levels were also measured quantitatively by the same EIA system with some modification. Total agreement of diagnosis by the on-farm EIA kit and by farrowing or abortion from 128 tested sows was found to be 92.2% accuracy (93.1% on pregnant diagnosis and 83.3% on non-pregnant diagnosis). It was concluded that the on-farm EIA blood progesterone test is a very useful method for monitoring the early pregnancy status of sows.

• 한국식품영양학회지
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• 제19권3호
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• pp.296-300
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• 2006

시유, 분유, 발효유에 존재하는 우유알레르겐인 ${\beta}$-Lactoglobulin (BLG)를 측정하기 위해 면역 blotting 및 간접결합면역 분석법(Competitive-indirect Enzyme linked Immunosorbent Assay; Ci-ELISA)으로 분석하였다. Immunoblotting 결과, 발효유에서는 우유 알레르기 환자의 IgE와의 반응이 나타나지 않았고 분유 및 시유에서는 약하지만 반응이 나타났다. 다클론 항체 및 우유 알레르기 환자의 IgE로 이들의 BLG 함량을 정량한 결과, BLG 함량은 시유가 가장 높았으며 발효유는 매우 낮았다. 이상의 결과에서 발효유에서는 가장 낮은 BLG 함량이 측정되어 우유 알레르기 환자에 대한 저알레르기원성 식품으로 사용될 수 있을 것으로 사료된다.

• 동의생리병리학회지
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• 제18권3호
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• pp.874-879
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• 2004

Alakaline phosphatase (ALP)- or horseradish peroxidase (HRP)-antibody conjugate was used frequently on the immunological detection methods such as enzyme-linked immunosobent assay (ELISA), immunobolt, immunohistochemistry. The classical enzyme-antibody coupling method by one-step (direction) injection of glutaraldehyde bring into being disadvantage such as low sensitivity of antigen detection because of homopolymers. This study was modified with the dialysis glutaraldehyde method to provide simple coupling through E-amino residues present in most protein. The dialysis glutaraldehyde coupling effects were better than the classical one-step glutaraldehyde injection in antigen detection of ELISA and immunobolt. Optimal dose of the dialysis glutaraldehyde solution was 0.10-0.25 %. This results suggest that the dialysis glutaraldehyde coupling method can readily applied to antigen detection of in vitro and in vivo.

• Journal of Animal Science and Technology
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• 제64권4호
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• pp.621-639
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• 2022

Cortisol and corticosterone, hormones traditionally considered biomarkers of stress, can be measured in fluid biomatrices (e.g., blood, saliva) from live animals to evaluate conditions at sampling time, or in solid biomatrices (e.g., hair, feather) from live or dead animals to obtain information regarding long-term changes. Using these biomarkers to evaluate physiological stress responses in domestic animals may be challenging due to the diverse characteristics of biomatrices for potential measurement. Ideally, a single measurement from the biomatrix should be sufficient for evaluating chronic stress. The availability of appropriate and cost-effective immunoassay methods for detecting the biomarkers should also be considered. This review discusses the strengths and limitations of different biomatrices with regard to ensuring the highest possible reliability for chronic stress evaluation. Overall, solid biomatrices require less frequent sampling than other biomatrices, resulting in greater time- and cost-effectiveness, greater ease of use, and fewer errors. The multiplex immunoassay can be used to analyze interactions and correlations between cortisol and other stress biomarkers in the same biomatrix. In light of the lack of information regarding appropriate biomatrices for measuring chronic stress, this review may help investigators set experimental conditions or design biological research.

• 한국환경농학회지
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• 제11권1호
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• pp.59-66
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• 1992

Endosulfan을 복잡한 시료조제과정이 필요치 않는 ELISA기법으로 분석법을 정립하고자 하였다. 항원조제를 위한 endosulfan alcohol의 합성은 86%의 수율을 보였고 endosulfan hemisuccinate는 HPLC로 그 합성을 확인하였다. Endosulfa-hemocyanin(KLH) 결합체의 수율은 3.57mg/ml였으며 이 항원은 1.1mg/ml로 희석하여 토끼에 대해 피하주사로서 면역시켰고 8주 후에 채혈된 항체 용액은 indirect ELISA로 1 : 24,000의 역가를 결정하였다. 또한 endosulfan-peroxidase 결합체는 약 40%의 결합수율인 2mg/ml로 조제되었으며 이 결합체는 높은 흡광도와 민감도를 나타내는 200ng/ml의 농도로 사용하였다. ELISA 분석에 의한 endosulfan의 검출한계는 5 ppb였으며 분해산물인 endosulfan ether와 endosulfan alcohol의 endosulfan 항체에 대한 민감도는 각각 57.14%, 105.20%였다. 유사한 유기염소계 농약들에 대한 교차반응성은 captan이 2.2%로 가장 낮게 나타났고 Captafol이 29.33%로 가장 높게 나타났다. ELISA에 의한 잔류 분석을 토양과 사과에서 응용 시험한 결과 토양에서는 10 ppb와 100 ppb에서 각각 97.45%, 98.40%의 회수율을 보였고 사과에서는 각각 98.40%, 100.55%의 회수율을 나타냈다.

• Journal of Microbiology and Biotechnology
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• 제13권4호
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• pp.509-516
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• 2003

A model system for the immnunochemical detection of Escherichia coli O157:H7 using a combined immunomagnetic separation (IMS) and test-strip liposome immunoassay (LIA) procedure was developed. Immunomagnetic beads coated with anti-E. coli O157 IgG antibodies were used to separate the E. coli O157 (including the H7 serotype) from culture. Immunoliposomes, whose surface was conjugated to goat anti-E. coli O157:H7 IgG and which encapsulated the marker dye, sulforhodamine B, were used as a detection label. The test strip, onto which antibodies to goat IgG were immobilized, was the immunosensor capturing immunoliposomes that did not bind to E. coli O157:H7 on the immunomagnetic bead-E. coli O157:H7 complexes. In experiments, pure cell culture suspensions of $10^5 E.$ coli O157:H7 organisms per ml produced a measurable signal inhibition, whereas a weak yet detectable signal inhibition occurred with $10^3CFU/ml$. The inhibition signals increased, when the incubation time for IMS was extended to 90 min and higher IgG-tag density (0.4mol%) was used on the liposomes. With 0.2 and 0.4mol% IgG-tagged liposomes, the IMS-LIA procedure showed more improved signal inhibitions than those of a direct (no IMS) LIA. The combined assay, which measures the instantaneous signal from immunoliposomes, can be completed within 90 min, making it significantly faster than conventional plating methods and enzyme-linked immunosorbent assay (ELISA). Accordingly, it is quite feasible to use the combined immunoassay format of IMS and dye-loaded immunoliposomes for the detection of E. coli O157:H7.

• Journal of Microbiology and Biotechnology
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• 제31권5호
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• pp.705-709
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• 2021

Porphyromonas gingivalis (P. gingivalis) is a major bacterial pathogen that causes periodontitis, a chronic inflammatory disease of tissues around the teeth. Periodontitis is known to be related to other diseases, such as oral cancer, Alzheimer's disease, and rheumatism. Thus, a precise and sensitive test to detect P. gingivalis is necessary for the early diagnosis of periodontitis. The objective of this study was to optimize a rapid visual detection system for P. gingivalis. First, we performed a visual membrane immunoassay using 3,3',5,5'-tetramethylbenzidine (TMB; blue) and coating and detection antibodies that could bind to the host laboratory strain, ATCC 33277. Antibodies against the P. gingivalis surface adhesion molecules RgpB (arginine proteinase) and Kgp (lysine proteinase) were determined to be the most specific coating and detection antibodies, respectively. Using these two selected antibodies, the streptavidin-horseradish peroxidase (HRP) reaction was performed using a nitrocellulose membrane and visualized with a detection range of 10³-10⁵ bacterial cells/ml following incubation for 15 min. These selected conditions were applied to test other oral bacteria, and the results showed that P. gingivalis could be detected without cross-reactivity to other bacteria, including Streptococcus mutans and Escherichia fergusonii. Furthermore, three clinical strains of P. gingivalis, KCOM 2880, KCOM 2803, and KCOM 3190, were also recognized using this optimized enzyme immunoassay (EIA) system. To conclude, we established optimized conditions for P. gingivalis detection with specificity, accuracy, and sensitivity. These results could be utilized to manufacture economical and rapid detection kits for P. gingivalis.

• Bulletin of the Korean Chemical Society
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• 제23권4호
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• pp.605-609
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• 2002

A competitive enzyme-linked immunosorbent assay (ELISA) was developed for the quantitative detection of organophosphorus insecticide cyanophos. An analogue (hapten) of cyanophos was synthesized and was coupled to BSA to produce polyclonal antibodi es from rabbits. The antisera were screened against another hapten coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigen-coated ELISA was developed, which showed an I50 of 310 ng/mL with the detection limit of 20 ng/mL. The antibodies showed negligible cross-reactivities with other organophosphorus pesticides except for parathion-methyl, which makes the assay suitable for the selective detection of cyanophos.