• Title/Summary/Keyword: enzyme-linked immunosorbent assay(ELISA)

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The Production of Antibody Against Sterigmatocystin Produced by Aspergillus vericolor (Aspergillus vericolor가 생산하는 sterigmatocystin에 대한 항체생산)

  • 윤원한;하우송;강진순;여명재;전향숙;정덕화
    • Journal of Food Hygiene and Safety
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    • v.10 no.1
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    • pp.1-6
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    • 1995
  • In order to establish the enzyme linked immunosorbent assay(ELISA) of sterigmatocystin produced by Aspergillus versicolor, we experimented and obtained following results. Two of three rabbits which had been immunized with sterigmatocystin-hemiacetal-BSA produced antibodies against sterigmatocystin at 15 weeks. The produced antibodies were specific for sterigmatocystin and sterigmatocystin-hemiacetal but didn't cross react with other sterigmatocystin analogues in a significant degree. DMF : 4% KC1 (18 : 2) mixed solution was most effective to dissolve sterigmatocystin. For the preparation of sample solution to determine sterigmatocystin by ELISA, sample was extracted with CHC13 and dried, than the dried sample was redissolved with 100 ${mu}ell$ DMF + 4% KC1 mixture. 10~1,000 ng/$m\ell$ level of standard sterigmatocystin could be applied to the established ELISA. When artifically contaminated rice were assayed by the ELISA, the average recovery of sterigmatocystin spiked to 25~500 ng/g was 109% (97~116%), and mean interwell coefficient of variation was 21% (11~28%).

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Development of an ELISA for the Organophosphorus Insecticide Chlorpyrifos

  • Cho, Young Ae;Lee, Hye-Sung;Park, Eun-Yeong;Lee, Yong-Tae;Hammock, Bruce D.;Ahn, Ki-Chang;Lee, Jae-Koo
    • Bulletin of the Korean Chemical Society
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    • v.23 no.3
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    • pp.481-487
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    • 2002
  • A selective enzyme-linked immunosorbent assay (ELISA) for the insecticide chlorpyrifos was developed. Four haptens for chlorpyrifos were synthesized and two of them were used as immunogens after coupling to keyhole limpet hemocyanin by two differe nt approaches. Rabbits were immunized with either of them and the sera were screened against 4 haptens coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigencoated ELISA was developed, which shows an I50 of 160 ppb with a detection limit of 10 ppb. An antibody-coated ELISA was also developed, which shows an $I_{50}$ of 20 ppb with a detection limit of 0.1 ppb. The antibodies showed negligible cross-reactivity with other organophosphorus pesticides except for insecticides chlorpyrifos-methyl and bromophos-ethyl, which makes these assays suitable for the selective detection of chlorpyrifos.

Feminization and reduction of testicular weight in mouse sparganosis

  • Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.44 no.2 s.138
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    • pp.167-169
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    • 2006
  • After infection of male mice with the plerocercoids (spargana) of Spirometra mansoni, serum levels of estrogen and testicular weight were analyzed by enzyme-linked immunosorbent assay (ELISA) and weighing machine, respectively. The serum level of estrogen increased progressively in infected mice compared with normal controls, whereas the testicular weight of infected mice decreased significantly (P < 0.05). These results suggest that certain substances from spargana change the steroid hormone metabolisms in the host by unknown pathways, and chronic infection may contribute to change of the function of steroid hormone target organ, i.e., testis, in male mice.

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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    • v.6 no.2
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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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Evaluation on diagnostic efficiency of the standard tube agglutination test for bovine brucellosis (소 브루셀라병 표준시험관응집반응법 진단효율 평가)

  • Sung, So-Ra;Kim, Ji-Yeon;Her, Moon;Lee, Kichan;Gu, Jeong-Hui;Kang, Sung-Il;Lee, Hyang-Keun;Kim, Suk-Mi;Jung, Suk-Chan
    • Korean Journal of Veterinary Service
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    • v.35 no.4
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    • pp.269-273
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    • 2012
  • A confirmatory serological test, the standard tube agglutination test (STAT) is evaluated for the diagnostic efficiency in brucellosis Korea. A total of 345 bovine samples were collected from regional veterinary branch under national brucellosis monitoring program from January 2010 to June 2012 in Korea. These samples were diagnosed as suspected serum and brucellosis positive by the Rose Bengal test (RBT) and the STAT, respectively. The STAT was compared and evaluated with three serological test such as the indirect-enzyme linked immunosorbent assay (I-ELISA), competitive-enzyme linked immunosorbent assay (C-ELISA) and fluorescence polarisation assay (FPA) prescribed for international trade by OIE. Among the 345 bovine serum samples, 302 (87.5%) were diagnosed as positive in the STAT, while 215 (62.3%), 223 (64.6%) and 194 (56.2%) serum samples were diagnosed as positive for brucellosis in the I-ELISA, C-ELISA and FPA, respectively. The STAT showed quite high positive results as compared with three prescribed tests of OIE. FPA, I-ELISA and C-ELISA have shown 60.6%, 64.9% and 67.2% correlation, respectively as compared to the STAT. However correlations of three prescribed tests ranged high 84.1~97.7%. Especially, correlation between I-ELISA and C-ELISA is quite high, 97.7%. These results suggest that the STAT has shown many false-positive reactions. Therefore, additional serological test, such as ELISAs and FPA, would be necessary to adopt as a confirmatory test in the national surveillance program of bovine brucellosis in Korea.

옥수수중 Deoxynivalenol의 검출을 위한 효소면역측정법의 개발

  • Lee, Hyang-Burm;Shon, Dong-Hwa;Kosaka, Kunio;Ueno, Yoshio
    • Microbiology and Biotechnology Letters
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    • v.25 no.4
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    • pp.414-419
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    • 1997
  • In order to develop an enzyme-linked immunosorbent assay (ELISA) for deoxynivalenol (DON) in com, we produced a specific monocl- onal antibody and established ELISA conditions. After the spleen cells from mice immunized with DON-bovine serum albumin conjugate were fused with S$_{p}$2/0 myeloma cells, a hybridoma cell 3G7 producing anti-DON antibody was screened by ELISA. From the standard curve of competitive direct ELISA (cdELISA) using 3G7 monoclonal antibody and DON-HRP conjugate, the detection range of DON showed 3-3,000 ng/ml (ppb). The monoclonal antibody showed some cross-reactivities against DON analogues such as 15 acetyl-DON (110%), nivalenol (5.0%), 3 acetyl-DON (1.7%), fusarenon-x (0.72%), and T-2 (0.59%). When the cdELISA was applied to the spiked coms after extracting with 60% methanol and diluting 5- fold with washing buffer, the assay recoveries of DON were 313, 163, 106, and 88.9% (av., 168%) in the levels of 200, 600, 2,000, and 6,000ng/g, respectively. For the quantitation of DON in coms, 30 samples kept under two different storage conditions of cold and room temperature were assayed by cdELISA. The mean detection concentrations were 595 (detection range, 0-2,750) and 2,448 (detection range, 0-4,500) ppb, respectively.

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Comparison of a new ELISA with other serodiagnostic tests for bovine brucellosis (소 브루셀라병의 혈청학적 진단법 비교실험)

  • Hur, Jin;Kakoma, Ibulaimu;Jeong, Jae-Myong;Lee, Hyeon-Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.385-391
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    • 2007
  • A novel enzyme linked immunosorbent assay (ELISA) is described and compared with other established serologic tests for bovine brucellosis, namely the rose bengal test (RBT), the complement fixation test (CFT), and the tube agglutination test (TAT) approved and used in Korea. A total of 109 bovine serum samples were tested using all the 4 assays and analyzed as to specificity, sensitivity, reproducibility and predictive value. The ELISA showed 100% agreement with the CFT. The least agreement between ELISA was observed with the TAT. The agreement between the ELISA and RBT was not significantly different from that observed between the CFT and the ELISA. It is concluded that the new assay would be a good candidate for routine serologic survey for brucellosis in Korea. A protocol combining the ELISA and the CFT would increase the power for detection of serologically positive individuals and herds.

Comparison between indirect immunofluorescent antibody (IFA) test and enzyme-linked immunosorbent assay (ELISA) for the detection of antibody to porcine reproductive and respiratory syndrome virus(PRRSV) (돼지 생식기호흡기증후군 바이러스항체 검색에 있어 간접형광항체법(IFA) 과 효소면역법(ELISA)의 진단효율 비교)

  • Park, Choi-kyu;Lyoo, Young-soo;Lee, Chang-hee;Jung, Jong-wook
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.314-318
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    • 1998
  • An establishment of effective control measures to PRRSV infection in swine industry depends on a sensitive and specific diagnosis to detect either viral antigen and/or antibodies to PRRSV. Several diagnostic methods are available to detect antibodies against PRRSV, including IPMA, IFA and ELISA tests have been successfully developed. Sensitivity of the indirect immunofluorescent assay in MA-104 cells using Korean field isolate PL96-1 was superior to that of VR-2332 and field isolate PL96-2. Sensitivity and specificity of the IFA test with PL96-1 were comparable to those of commercial ELISA test kit but ELISA test was more sensitive for the detection of declining antibodies to PRRSV in finishing pigs. In this study we concluded that IFA and ELISA test could be utilized to detect antibodies to PRRSV and the results generated from these two tests were comparable and there were no significant difference between these two tests.

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Development of an Indirect Enzyme-Linked Immunosorbent assay for Rapid Detection of Adulteration of Food Allergen Mackerel in Processed Marine Foods (수산가공식품 중 알레르겐 고등어 혼입여부 신속 검출을 위한 간접효소면역분석법의 개발)

  • Lee, Jeong-Eun;Kim, Ah-Yoon;Kim, Sol-A;Kim, Hyo-In;Park, Ji-Hye;Shim, Won-Bo
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.185-192
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    • 2018
  • The purpose of this study was to develop an indirect enzyme-linked immunosorbent assay (indirect ELISA) based on a monoclonal antibody (MAb) that is specific to mackerel thermal stable-soluble protein (TSSP), that can be used for the rapid detection of mackerel in processed marine foods. Among the four MAbs (3A5-1, 2, 9, and 12) developed in previous studies, the 3A5-2 MAb that showed high specificity and sensitivity were selected and used to develop the indirect ELISA method. The detection range of the indirect ELISA was 0.02%-0.001% and the detection limit of 0.001% was shown. No cross-reaction to other marine products and food ingredients was observed by the indirect ELISA. Processed marine foods containing mackerel with ${\geq}0.3$ O.D. value at 405 nm were estimated as positive samples by the indirect ELISA. Therefore, the indirect ELISA can be used as a rapid and sensitive method to identify mackerel authenticity and adulteration in processed marine foods.

효소면역측정법에 의한 우유중의 Aflatoxin M$_{1}$ 분석

  • 손동화;임선희;이인원
    • Microbiology and Biotechnology Letters
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    • v.24 no.5
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    • pp.630-635
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    • 1996
  • For a survey of the occurrence of aflatoxin M$_{1}$ (AFM$_{1}$) in domestic cow's milk, we developed an enzyme-linked immunosorbent assay (ELISA) system, and quantitated the toxin in cow's milk. In order to produce specific antibodies AFM, conjugated to bovine serum albumin (AFM$_{1}$-BSA) and Freund's adjuvant were immunized subcutaneously to rabbits. By use of the antiserum showing the highest titer and AFB$_{1}$-HRP conjugate, we established a competitive direct ELISA (cdELISA) for AFM$_{1}$, whose detection limit was 0.003 ppb. The cross-reactivities of the antiserum against aflatoxin M$_{1}$ M$_{2}$, B$_{1}$, B$_{2}$, G$_{1}$, G$_{2}$, B$_{2a}$, and G$_{2a}$, were 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, and 0%, respectively. When the cdELISA was applied to the cow's milk spiked with AFM$_{1}$ and followed by cleanup with C$_{18}$ cartridge, the mean recovery of the assay was 104% (mean of CV, 6.4%) in the final concentration of 0.01-1 ppb (10-1, 000 ppt). When cow's milk samples gathered from markets and farms were assayed by the cdELISA, the mean concentration and SD of AFM$_{1}$ was 80.4 $\pm$ 55.0 ppt (n=64; range, 5.6-280 ppt).

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