• Title/Summary/Keyword: ELISA.

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Development of an ELISA for the Organophosphorus Insecticide Isofenphos

  • Park, Han-Jin;Park, Won-Chul;Jung, Tae-Owan;Rha, Choon-Sup;Lee, Yong-Tae
    • Bulletin of the Korean Chemical Society
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    • v.23 no.4
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    • pp.599-603
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    • 2002
  • A selective enzyme-linked immunosorbent assay (ELISA) for the insecticide isofenphos was developed. Three different analogues (haptens) of isofenphos were synthesized and were coupled to carrier proteins through the pesticide thiophosphate group t o use as immunogens or coating antigens. Rabbits were immunized with one of the haptens coupled to BSA for production of polyclonal antibodies and the sera were screened against each of the other two haptens coupled to ovalbumin (OVA). Using the sera of highest specificity, an antigen-coated ELISA was developed, which showed an I50 of 96 ng/mL with the detection limit of 2 ng/mL. The antibodies showed negligible cross-reactivity with other organophosphorus pesticides and the phenol metabolite of isofenphos, which makes the developed assay suitable for the selective detection of isofenphos. An antibody-coated ELISA was also developed, which showed an I50 of 580 ng/mL with a detection limit of 70 ng/mL.

Serosurvey for antibodies against brucellosis in pigs (돼지 brucellosis에 대한 항체가 조사)

  • Hur, Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.153-157
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    • 2011
  • In order to investigate serum antibodies for detection of brucellosis in pigs, a total of 1208 sera were tested by Rose Bengal test (RBT), the standard tube agglutination test (STAT) and competitive ELISA (cELISA). The sera were collected from pigs of Gyeonggi, Chungnam, Chungbuk, Jeonnam and Jeonbuk, provinces during the period 2002 to 2004. All the sera were screened by RBT, and were confirmed by STAT and cELISA. Among 1208 sera, 26 sera (2.2%) were positive in screening test. All the 26 positive sera were positive by STAT, while all the sera were negative by cELISA. On the basis of this study, farmed pigs may be exposed to Brucella species. Furthermore, these results suggest that establishment of diagnoses for detection of porcine brucellosis is necessary.

Evaluation in ELISA for the Residue Analysis of Metalaxyl in Crops (작물체중 Metalaxyl 의 잔류분석을 위한 ELISA의 응용)

  • Lee, Kang-Bong;Im, Geon-Jae;Jung, Young-Ho;Suh, Yong-Tack
    • Korean Journal of Environmental Agriculture
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    • v.14 no.1
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    • pp.23-27
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    • 1995
  • The cross-reactivities of antibody derived from rabbit immunized with metalaxyl-HSA conjugate were identified to 44% and 28% for metalaxyl acid and metolachlor respectively. The detection range of metalaxyl in competitive ELISA was 5 ppb to 5 ppm. The recoveries of metalaxyl in ELISA for 6 crops; potato, sesame, pepper, cabbage, cucumber and onion was ranged 77.5-103.6%. In this test, CV% were calculated below 10% except for sesame sample as shown 21.5%.

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Diagnosis of canine brucellosis using recombinant ribosomal protein L7/L12 (재조합 리보솜 단백질 L7/L12을 이용한 개 브루셀라병의 진단)

  • Lee, Hyang-Keun;Kim, Jong-Wan;Ha, Yun-Mi;Her, Moon;Kim, Ji-Yeon;Lee, Kichan;Kang, Sung-Il;Jung, Suk-Chan
    • Korean Journal of Veterinary Research
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    • v.52 no.1
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    • pp.25-31
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    • 2012
  • Brucella (B.) canis is mainly transmitted by direct or indirect contact with aborted fetuses and placenta. It's also known to be able to infect human, which likely results in providing veterinarians and companion animal owners for infectious risk. To develop diagnostic ELISA, we cloned and expressed rp1L gene of B. canis, which encodes the ribosomal protein L7/L12. Using this purified recombinant protein, indirect-ELISA (iELISA) was evaluated using 78 positive and 44 negative sera. The sensitivity and the specificity of iELISA were 94% and 89%, respectively. The results indicated that indirect-ELISA using recombinant ribosomal protein L7/L12 may be useful for diagnosis of canine brucellosis.

Development of a Blocking ELISA for Measuring Rabies Virus-specific Antibodies in Animals

  • Yang, Dong-Kun;Kim, Ha-Hyun;Ryu, Jieun;Gee, Mi-ryun;Cho, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.269-276
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    • 2018
  • Rabies virus (RABV)-specific antibodies in animals and humans are measured using standard methods such as fluorescent antibody virus neutralization (FAVN) tests and rapid fluorescent focus inhibition tests, which are based on cell culture systems. An alternative assay that is safe and easy to perform is required for rapid sero-surveillance following mass vaccination of animals. Two purified monoclonal antibodies (4G36 and B2H17) against RABV were selected as capture and detection antibodies, respectively. A genetically modified RABV, the ERAGS strain, was propagated and concentrated by polyethylene glycol precipitation. Optimal conditions for the RABV antigen, antibodies, and serum dilution for a blocking enzymelinked immune sorbent assay (B-ELISA) were established. We evaluated the sensitivity, specificity, and accuracy of the B-ELISA using serum samples from 138 dogs, 71 raccoon dogs, and 25 cats. The B-ELISA showed a diagnostic sensitivity of 95.8-96.3%, specificity of 91.3-100%, and accuracy of 96.0-97.2% compared to the FAVN test. These results suggest that the B-ELISA is useful for sero-surveillance of RABV in dogs, raccoon dogs, and cats.

Enzyme-linked Immunosorbent Assays (ELISA) and Immunochromatography Assays (ICG) for Analysis of Vitellogenin in the Scorpion Fish Sebastiscus marmoratus (쏨뱅이(Sebastiscus marmoratus)의 Vitellogenin 분석을 위한 효소면역측정법(ELISA) 및 면역크로마토그래피분석법(ICG) 개발)

  • Yeo, In-Kyu;Lim, Yoon-Kyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.4
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    • pp.459-465
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    • 2015
  • We tested biomarker systems [enzyme-linked immunosorbent assay (ELISA) and immunochromatography assay (ICG) kits] for the screening of endocrine-disrupting chemicals in contaminated environments using antibodies resulting from $17{\beta}$-Estradiol-induced vitellogenin (Vtg) in the wild scorpion fish Sebastiscus marmoratus. Monoclonal antibodies of two clones (S28 and S15) were used as capture and tracer antibodies for ELISA and ICG assays. ELISA detected Vtg at levels greater than $0.1{\mu}g/mL$, while ICG detected Vtg at levels greater than $1{\mu}g/mL$. However, the ICG system was able to detect antibodies from $17{\beta}$-Estradiol-induced Vtg serum that had been diluted 1,000 times. Our results suggest that previously developed biomarker assays can be used as detection systems to detect known endocrine-disrupting chemicals in contaminated environments, and to measure their activity.

Development and Evaluation of a Competitive Enzyme Immunoassay for the Detection of Antibodies to Treponema pallidum (경쟁적 효소면역 측정법을 이용한 Treponema pallidum 항체 진단시약의 개발 및 평가)

  • 김병문;이정환;정문섭;김승철;이미용;이성희;김원배
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.344-348
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    • 1999
  • A competitive enzyme-linked immunosorbent assay(ELISA) for the detection of antibodies to Treponema pallidum(T.pallidum) was developed and evaluated. T.apllidum lysate was immobilized on the surface of microplate wells and horseradish peroxidase labeled human anti-T.pallidum lysate was immobilized on the surface of microplate wells and horseradish peroxidase labeled human anti-T.pallidum was prepared and used as a tracer. The performance of the competitive ELISA was evaluated by using different specimens. The competitive ELISA showed a sensitivity of 100% in a performance panel consisting of serum and plasma with anti-T.pallidum reactivity ranging from negative to strong positive by FTA-ABS test system and 120 plasma samples positive by TPHA. The specificity of the competitive ELISA was 100% in 1,200 plasma samples collected from healthy seronegative blood donors. These results suggest that the competitive ELISA provides an excellent assay method for the detection of antibodies to T.pallidum, and may be particularly useful for serological blood screening of syphilis.

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Epidemiological characteristics of classical swine fever outbreak at Jeonbuk area in 2003 (전북지역에서 발생한 돼지콜레라의 역학적 특성)

  • Eum Sung-Shim;Lee Jeoung-Won;Seo Lee-Won;Bea Joung-Jun;Joung Dong-Suk
    • Korean Journal of Veterinary Service
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    • v.27 no.3
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    • pp.239-247
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    • 2004
  • Classical swine fever (CSF) was confirmed in 19 herds in Jeunbuk provence (Iksan, Gimje, Wanju, Buan, and Jangsu) in Korea between March and May, 2003 and 10,263 pigs were slaughtered. Pigs contacted with CSF virus in primary outbreak farm show fever, reduced appetite, arched back and chill in company with sever respirative sign and then most infected farms also were observed to fever, reduced appetite, sudden death, and leukopenia (101 pigs). In order to detecting infectious pig with CSF virus, A total of 555 pigs were inspected in 65 herds and blood samples were collected and serological test (ELISA), antigen ELISA, and reverse transcription-polymerase chain reaction (RT-PCR) had been done. Positive rate were $74\%$ (410 pigs) in antibody ELISA, $2\%$ (11 pigs) in antigen ELISA and $33\%$ (182 pigs) in RT-PCR, respectively. As shown that the RT-PCR was useful than the ELISA for determining CSF virus in blood, meat, and other organs.

Studies on serological tests for pullorum disease (추백리의 혈청학적 진단법에 관한 연구)

  • 김정태;심항섭;김태종;고태오;우종태;유기승;박유순
    • Korean Journal of Veterinary Service
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    • v.21 no.3
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    • pp.313-323
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    • 1998
  • In order to establish a sensitive and specific diagnostic method for detection of antibody to Salmonella pullorum, a enzyme-linked immunosorbent assay(ELISA) was designed and standardized. The diagnostic efficacy of the established ELISA was compared with that of the serum plate agglutination test and immunodiffusion test for pullorum disease. 1. The chicken hyperimmune sera to Salmonella pullorum, S gallinarum, S typhimurium and S typhi were shown the cross reaction to S pullorum antigen by serum plate agglutination test. 2. When compared the cross reaction titer of microplate agglutination test for chickens hyperimmune sera, it was found that the titer were 64 in S pullorum, 32 in S gallinarum, 4 in S typhimurium and 8 in S typhi, respectively. 3. When compared the specificity of various antigen(HA, EA, PA and SA) by the immunodiffusion test, the most suitable antigen was phenol-treated bactrium. 4. The optimal concentration of S pullorum antigen for ELISA was 1 : 160 dilution of bacterium. 5. The efficacy of the ELISA for detection of S pullorum antibody was compared with serum Plate agglutination test and immunodiffusion test in chickens infected with S pullorum. The antibody was first detected at 6 days after infection using three tests examined. The antibody was alldetected at 9 days by ELISA, at 12 days by serumplate agglutination test, at 15 days by immunodiffusion test.

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Characteristics and application of monoclonal antibody to progesterone II. Development of progesterone enzyme-linked immunosorbent assay(ELISA) (Progesterone의 단크론성 항체에 관한 특성 및 활용에 관한 연구 II. ELISA 기법의 개발)

  • Kang, Chung-boo;Kim, Jong-shu
    • Korean Journal of Veterinary Research
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    • v.31 no.4
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    • pp.403-409
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    • 1991
  • This experiment was carried out to develop a sensitive, rapid, solid-phase microtitre plate assay of progesterone using the monoclonal antibody to this hormone. Monoclonal antibody to progesterone was much higher titre and binding affinity about 10 times than conventional polyclonal antibody to progesterone. Dot-blot analysis of monoclonal antibody revealed a single precipitation band when reacted with anti-mouse IgM and anti-mouse K. A competitive reaction was used with a reaction time of 2 hours. The standard dose-response curve was linear through 1,000pg/well. This ELISA system approach is applicable to evaluation for the rapid assessment of luteal function and reproductive status in both clinical and research in a wide variety of species.

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