• Title/Summary/Keyword: Enzyme immunoassay

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Novel Liposome Immunoassay for Detection of Ultratrace Amount of Bioactive Substances : an Assay for Insulin

  • Lim, Soo-Jeong;Kim, Chong-Kook
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.281-281
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    • 1996
  • The immunoassay method is frequently used for the identification and quantitation of ultratrace amount of bioactive substances. Homogeneous liposome immunoassays, which can avoid the use of radioisotopes and separation steps, have recently been reported in many publications. Cytolysin-mediated liposome immunoassay using melittin ever been studied but showed limited applications. Here, we designed a homogeneous liposome immunoassay using Clostridium perfringens phospholipase C (PLC), an enzyme which catalyzes the hydrolysis of phosphatidylcholine in biological membranes, as a cytolysin.

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Development of an enzyme immunoassay for detection of Escherichia coli O157 in meat (식육중 Escherichia coli O157 검출을 위한 enzyme immunoassay 기법 개발)

  • Jung, Byeong-yeal;Jung, Suk-chan;Cho, Dong-hee;Kim, Jong-yeom;Park, Yong-ho;Shin, Sang-jae;Kim, Sung-guk;Kim, Bong-hwan
    • Korean Journal of Veterinary Research
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    • v.38 no.4
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    • pp.745-750
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    • 1998
  • A sensitive and rapid enzyme immunoassay(EIA) to detect Escherichia coli O157 in ground beef was developed by using a sandwich type assay with polyclonal antibodies to E coli O157. E coli O157 in ground beef could be detected within 15hr, including incubation for 12hr in enrichment broth and 3hr in immunoassay. The EIA could detect $1.3{\times}10^5$ cells of E coli O157/g of ground beef without enrichment. The lowest limit of detection was 0.23 E coli O157 per g of meat after enrichment. Confirmation was required in the positive specimens in the EIA by culture method even though the negative specimens were not. These results suggested that the immunoassay could be a very efficient method for the screening E coli O157 in meat.

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Polymerase Chain Reaction Detection of Mycobacterium tuberculosis and Fine Needle Aspiration Cytology for the Diagnosis of Tuberculous Lymphadenitis (결핵성 림프절염의 진단를 위한 세침흡인 세포검사 및 중합효소연쇄 반응과 효소면역법을 이용한 Mycobacterium tuberculosis의 검출)

  • Kim, Joo-Heon;Kim, Nam-Hoon;Kang, Dong-Wook;Park, Mee-Ja;Moon, Sang-Kyoung;Yu, Tae-Cho;Jang, Eun-Ju
    • The Korean Journal of Cytopathology
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    • v.12 no.1
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    • pp.25-30
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    • 2001
  • Tuberculous lymphadenitis is not uncommon in Korea. Therefore, an inexpensive, safe and rapid method is needed to diagnose the tuberculous lymphadenitis. Flne needle aspiration cytology Is a good method for this purpose, but has several limitations in the diagnosis of tuberculous lymphadenitis, especially when the presence of acid-fast bacilli is not proved. To evaluation the usefulness of the polymerase chain reaction with enzyme immunoassay technique in the detection of Mycobacterium tuberculosis (M. tuberculosis) In the cervical Iymph node asplrates, the authors performed fine needle aspiration cytology and M. tuberculosis PCR with enzyme immunoassay for mycobacterial DNA sequences from 15 cases of the fine needle aspirates. Cytomorphologically, the cases were categorized into three types: predominantly necrotic materials; typical epithelioid cell granulomas with or without slant cells and caseous necrosis; and non-tuberculous lesions, such as reactive lymphadenitis, abscess, metastatic carcinoma and malignant lymphoma. M. tuberculosis DNA was found in 8 of 15 cases by PCR with enzyme immunoassay. Negative findings on PCR were achieved in 7 cases, which revealed non-tuberculous tymphadenopathy. In conclusion, we suggest that M. tuberculosis PCR with enzyme immunoassay using the fine needle aspirates is a very useful tool for the diagnosis of tuberculous lymphadenitis.

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Development and validation of a simple, sensitive enzyme immunoassay for quantification of androstenedione in bull plasma

  • Mallick, Smrutirekha;Kumar, BS Bharath;Prakash, BS;Aggrawal, Anjali;Pandita, Sujata
    • Journal of Animal Science and Technology
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    • v.57 no.4
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    • pp.13.1-13.5
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    • 2015
  • As an alternative to radioimmunoassay a simple and highly sensitive enzyme immunoassay (EIA) was developed and validated for androstenedione quantification in plasma of Karan Fries bulls using second antibody coating technique. The wells of the microtitreplate were coated with affinity-purified goat immunoglobulin (antirabbit IgG) that binds the hormone specific antibody. The EIA was performed to analyze androstenedione directly in $40{\mu}l$ of bull plasma. The androstenedione standards ranged from 0.20 to 200 pg/$40{\mu}l$/well and the sensitivity of the assay was 5 pg/ml plasma. Serially diluted bull plasma containing high endogenous androstenedione showed good parallelism with bovine androstenedione standard curve. Intra- and inter-assay coefficients of variation (CV) were found to be 8 and 9%, respectively. Peripheral plasma androstenedione concentrations determined in young and adult bull samples ranged between 104-990 pg/ml and 184-2040 pg/ml, respectively.

Development of an enzyme immunoassay for determination of steroid hormones to improve the reproductive efficiency of domestic animals (가축(家畜)의 번식효율증진(繁殖效率增進)을 위한 steroid hormones 의 효소면역분석법(酵素免疫分析法) 개발(開發))

  • Choi, Han-sun;Kang, Byong-kyu
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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    • pp.611-615
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    • 1993
  • A rapid, solid-phase microtitre plate enzyme immunoassay(EIA) to determine the concentration of progesterone and testosterone in dairy cow is described. Both steroid hormones were analysed employing antibodies against $11{\alpha}$-hemisuccinate-progesterone bovine serum albumin and 4-androsten-$17{\beta}$-ol-3-one-carboxymethyloxime bovine serum albumin, respectively. as primary antibodies and sheep Ig G as secondary antibody. The conjugated used as labels for progesterone and testosterone was progesterone-$11{\alpha}$-hydroxy-hemisuccinate- horseradish peroxidase and 4 ${\alpha}$-androsten-$17{\beta}$-ol-3-hemisuccinate- horseradish peroxidase, respectively. Detection limit of microtitre plate EIA was 6.7 pg/well for progesteone and 1.0 pg/well for testosterone.

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Detection of Peanuts in Commercially Processed Foods by an Enzyme-Linked Fluorescent Immunoassay (Enzyme-linked fluorescent immunoassay에 의한 가공식품 중 땅콩의 검출)

  • Kim, Mi-Hye;Kim, Hyun-Jung;Shon, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.41 no.1
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    • pp.111-115
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    • 2009
  • In this study we analysed for peanuts in processed foods using an enzyme-linked fluorescent immunoassay (ELFA), and compared the results with labeled ingredients. Crude peanut protein (CPP) was immunized into rabbits to produce specific antibodies(Ab). A sandwich ELFA was established using anti-CPP Ab and Ab-horseradish peroxidase (HRP) conjugate. The cross-reactivities of the Ab toward CPP, peanuts, almonds, soybeans, and walnuts were 100, 9.8, $1.1{\times}10^{-2},\;4.4{\times}10^{-3}$, and 0%, respectively. The samples included 19 items consisting of biscuits, snacks, chocolates, and so on. The results from the sandwich ELFA showed that peanuts were contained in 7 of the processed food items, among which, 5 items were labeled as having peanuts present but 2 items were not. One of the 2 items that was peanut-detected but unlabeled was a biscuit labeled to contain almonds and assayed to contain $2.1{\times}10^{-3}%$ peanuts, which might have been due to the weak cross-reactivity of the Ab toward almonds. The other item was a snack labeled to contain soybeans and assayed to contain 0.098% peanuts, which might have been due to peanut cross-contamination during processing, since the crossreactivity of the Ab toward soybeans was very weak. These results suggest that ELFA is a good tool to detect peanuts in processed foods, and allergens in certain processed foods should be labeled correctly.

Analysis of Cow Hide Glue Binder in Traditional Dancheong by Enzyme-linked Immunosorbent Assay

  • Yu, Jia;Chung, Yong Jae
    • Journal of Conservation Science
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    • v.35 no.4
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    • pp.363-372
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    • 2019
  • Animal glue has been used as a binder in Dancheong since the Joseon dynasty. Binders play an important role in determining the physical characteristics of a painting layer. The analysis of binders can be used to identify the materials and techniques used in traditional Dancheong. Binders can be investigated using physicochemical component analyses methods such as gas chromatography/mass spectrometry, pyrolysis-gas chromatography/mass spectrometry, and fourier transform infrared spectroscopy, but the detection characteristics vary depending on the degradation properties of the pigment and binder. Therefore, cross-validation using a combination of physicochemical analysis and enzyme immunoassay is used to increase the reliability of the results. In this study, we present an enzyme-linked immunosorbent assay (ELISA) as an example of an enzyme immunoassay as a method for analyzing animal glue, a traditional binder used in Korea. The applicability of ELISA was tested using commercial animal glue, in addition to animal glue produced using a variety of extraction conditions. The animal glue was analyzed in a Noerok-additionally coated-replica sample to evaluate the possibility of analyzing the animal glue in a paint layer mixed with pigment. Based on the results, we performed an assay on the use of animal glue in the Dancheong sample of the temples of the Joseon dynasty, that are estimated to have been built in the 17th century.

Simple Electrochemical Immunosensor for the Determination of Rabbit IgG Using Osmium Redox Polymer Films

  • Choi, Young-Bong;Lee, Seung-Hwa;Tae, Gun-Sik
    • Journal of the Korean Electrochemical Society
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    • v.10 no.3
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    • pp.229-232
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    • 2007
  • An amperometric immunosensor for the determination of rabbit IgG is proposed. The immunoassay utilizes a screen-printed carbon electrode on which osmium redox polymer is electrodeposited. This immunoassay detects 0.1 ng/ml of rabbit IgG, which is ${\sim}10^2$ fold higher than the most sensitive enzyme amplified amperometric immunoassay. The assay utilizes a screen-printed carbon electrode which was pre-coated by a co-electrodeposited film of an electron conducting redox hydrogel and a rabbit IgG. The rabbit IgG in the electron conducting film conjugates captures, when present, the anti-rabbit IgG. The captured anti-rabbit-IgG is labeled with horseradish peroxidase (HRP) which catalyzes the two-electron reduction of $H_2O_2$ to water. Because the redox hydrogel electrically connects HRP reaction centers to the electrode, completion of the sandwich converts the film from non-electrocatalytic to electro-catalytic for the reduction of $H_2O_2$ to $H_2O$ when the electrode is poised at 200 mV vs. Ag/AgCl.