• Title/Summary/Keyword: Dot-ELISA

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Studies on the mycoplasmal pneumonia in slaughter pigs. 1. Seasonal detection by gross finding of lung lesion and dot-ELISA technique (도축돈의 마이코플라즈마성 폐렴에 관한 연구 1. 육안적 폐병변과 dot-ELISA에 의한 계절별 조사)

  • Lim, Young-Taek;Seok, Ho-Bong
    • Korean Journal of Veterinary Research
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    • v.42 no.2
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    • pp.219-224
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    • 2002
  • We report the seasonal prevalence of the mycoplasmal pneumoniae of swine (MPS) in slaughter pigs from July of 1999 to June of 2000. Gross finding of lung lesion observed and examined by dot-ELISA. In gross finding of lung lesion from 750 pig samples, 465 (62.0%) was MPS, and 129 (17.2%) was single or double infection with actinobacillosis and pasturellosis. However, 156 (20.8%) had no lesion. In seasonal detection, the prevalence was found to be winter (69.5%), autumn (63.5%), summer (60.0%) and spring (54.7%) in orderly frequency. In dot-ELISA, the result was showed the positive reaction (x16>titre) with 58.0% and negative (x4

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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Dot Blot Assay for Screening of Anti-hantavirus Antibodies by Using Nucleocapsid Protein of Hantaan Virus (한탄바이러스 핵단백질을 이용한 항 한타바이러스 항체 검색용 Dot Blot Assay)

  • Cho, Hae-Wol;Chung, Yeun-Jun;Kim, Chung-Lim;Ban, Sang-Ja;Nam, Jae-Hwan;Lee, Hyeong-Woo;Lee, Yoo-Jin;Kim, Eun-Jung
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.59-65
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    • 1996
  • For easy and rapid screening of hemorrhagic fever with renal syndrome (HFRS) without any laboratory equipment, dot blot enzyme immunoassay was developed and tried to detect anti-hantavirus antibodies. The nucleocapsid protein of Hantaan virus was isolated by affinity chromatography and used for making the dot strip. 28 of 29 Hantaan virus infected sera showed positive signals and 21 of 22 HFRS negative sera showed no positive signals. Anti-Seoul virus monoclonal antibody also exibited positive signal but the intensity of colorization was approximately 5 fold less than that of anti-Hantaan monoclonal antibody. The sensitivity of dot blot assay was equal or superior to indirect immunofluorescent assay (IFA) or ELISA test. Overall, the screening results with dot blot assay showed 92.2 % of concordance with IFA or ELISA test. This results suggests that dot blot assay could be applied a tool for easy and rapid screening of HFRS.

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Immunohistochemical Detection of the Grolwth Hormone-like Substance in Sparganum of Spirometra erinacei (고충(Sparganum)에서 성장호르몬 유사물질의 면역조직화학적 검출)

  • 김명옥;최완성김창환
    • The Korean Journal of Zoology
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    • v.35 no.2
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    • pp.173-182
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    • 1992
  • 흰쥐에 Spirometro rrinoce측 제3기 유충(고충 sparganum)이 감염되었을때 유충에서 생성된 성장호르몬 유사물질이 흰쥐 성장에 미치는 영향을 조사하기 위하여, 감염 기일이 경과에 따라 흰쥐의 뇌하수체와 유충의 체내에서 성장호르몬 분비세포를 면역조직화학염색으로 검색하였다. 유충의 표피와 표피성 근육층 및 근질 근육층에서 면역반응성을 띄는 성장 호르몬이 동정되었으며 감염시기의 경과에 따라 성장호르몬 분비세포의 수가 점차 증가하였다. 이와는 대조적으로 흰쥐의 뇌하수체에서는 성장호르몬 분비세포의 수가 감염 기일의 경과에 따라 정상대조군에 비하여 점차 감소 하였으며, 감염후 3개월이 경과되면 다시 증가하여 정상대조군의 수준으로 회복되었다. 또한 유충에 감염된 기일의 경과에 따른 횐쥐의 혈중 성장호르몬 농도변화를 dot-ELISA 방법으로 추정한 결깍 정상대조군의 성장호르몬의 양과 유의한 차이는 없었다. 결론적으로 유충의 체내에서 생성된 성장호르몬 유사물질이 전이숙주인 흰쥐의 성장을 유도할 것으로 사료된다.

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Identification of Chicken Pork and Beef Meats by Chicken Specific Antibody (계육 특이항체를 이용한 원료육 단백질의 검색)

  • 임태진
    • Food Science of Animal Resources
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    • v.18 no.2
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    • pp.149-156
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    • 1998
  • Chicken beef pork meats and isolated soy protein (ISP) were heated at 10$0^{\circ}C$ for 30min and then heat-resistant proteins were fractionated to examine cross-resistant protein from chicken meat but not with beef pork or ISP. Dot blotting using the polyclonal antibody showed that the sen-sitivity for detecting chicken meat was 1$\mu$m and antibody-antigen reaction was dose-dependant. Results of dot blotting analysis to compare the amount of chicken meat present in arket meat products(Kentucky Frank sausage;chicken meat 46.52% and pork 24.92% vs Bulgogi Ham;chicken meat 28.89% and turkey 31.44%)showed that the significant differences between two meat products in terms of chicken meat concentrations. Dose-dependant dot-blotting reaction was also observed in chicken meat samples with various dilution.

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Development of ELISA for brucella abortus RB51 I. Analysis on antigens of Brucella abortus RB51 by Westeren blot (부루세라 RB51의 ELISA 진단법개발 I. Westeren blot에 의한 Brucella abortus RB51균의 항원 분석)

  • Her, Moon;Cho, Dong-hee;Jung, Byeong-yeal;Cho, Seong-kun;Jung, Suk-chan;Kim, Ok-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.43-49
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    • 2001
  • As compared with reaction of antibody for sonicated antigen of Brucella abortus strain RB51 and 1119-3 by Western blot analysis, Brucella field positive sera was detected strong reaction at 40~80 kDa LPS of strain 1119-3, but detected very weak reaction at strain RB51 partly. Otherwise, as we analyzed major immunogen of RB51 by antisera bled periodically during 6 months after RB51 vaccination. we detected strong immunological reaction at 17, 18 and 8 kDa antigen of RB51. Especially, reaction of 8 kDa antigen by Western blot coincided with reaction of dot-blot assay in RB51-antibody detection method. We also compared with reaction of field sera by STAT(standard tube agglutination test), dot-blot assay and Western blot (reaction of 8 kDa antigen of strain RB51). 16 sera of 4~5 months after RB51 vaccination are all negative by STAT, and 12 field brucellosis positive serum are all positive, and also 12 of 16 sera vaccinated RB51 are positive by dot-blot assay and reaction of 8kDa antigen by Western blot. but 1 of 15 Brucellosis negative sera reacted nonspecifically dot-blot assay.

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Antiserum Preparation of Recombinant Sweet Potato Latent Virus-Lotus (SPLV-Lotus) Coat Protein and Application for Virus-Infected Lotus Plant Detection

  • He, Zhen;Dong, Tingting;Chen, Wen;Wang, Tielin;Gan, Haifeng;Li, LiangJun
    • The Plant Pathology Journal
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    • v.36 no.6
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    • pp.651-657
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    • 2020
  • Lotus is one of the most important aquatic vegetables in China. Previously, we detected sweet potato latent virus from lotus (SPLV-lotus) and found that it has highly significant sequence diversity with SPLV-sweet potato isolates (SPLV-sp). Here, we developed serological methods for the detection of SPLV-lotus in Chinese lotus cultivation areas. Based on the high sensitivity of SPLV-lotus coat protein antiserum, rapid, sensitive and large-scale diagnosis methods of enzyme-linked immunosorbent assay (ELISA) and dot blot in lotus planting area were developed. The established ELISA and dot blot diagnostic methods can be used to detect SPLV-lotus from samples successfully. And our results also showed that the SPLV-lotus and sweet potato isolates appeared clearly distinction in serology. Our study provides a high-throughput, sensitive, and rapid diagnostic method based on serology that can detect SPLV on lotus, which is suggested to be included in viral disease management approach due to its good detection level.

Development of a Disperse Dye Immunoassay Technique for Detection of Antibodies against Neospora caninum in Cattle

  • Selahi, Fatemeh;Namavari, Mehdi;Hosseini, Mohammad Hossein;Mansourian, Maryam;Tahamtan, Yahya
    • Parasites, Hosts and Diseases
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    • v.51 no.1
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    • pp.129-132
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    • 2013
  • In this study a disperse dye immunoassay method was standardized and evaluated for detection of antibodies against Neospora caninum in cattle. Sera from 150 cattle with a recent history of abortion were collected and tested by commercial ELISA kit and a standardized in-house dye immunoassay system. The positivity rate for the sera used in this study was 34.6% for the disperse dye immunoassay (DDIA) compared to 32% obtained by ELISA kit. This study showed no significant difference between DDIA and ELISA. The results indicated that the DDIA provide an economic, simple, rapid and robust test for detection of N. caninum infection in cattle.

Development of Rapid Diagnostic Kit for Identification of Hanwoo (Korean Native Cattle) Brand Meat by Detecting BIO-TAG

  • Baek, Kyung Hoon;Park, Sung Kwon;Lee, Myung Hoon;Kim, Sung Il;Cho, Soo Hyun;Choi, Chang Bon
    • Food Science of Animal Resources
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    • v.34 no.3
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    • pp.339-345
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    • 2014
  • This study was performed to develop a rapid immuno-assay kit, by using a specific antigen to detect Hanwoo brand meat. We selected a synthetic antigen specific to our target antibody, named BIO-TAG (Tyr-D-Ala-Phe), by utilizing a computer-based analysis and literature review. BIO-TAG tagged with adjuvant was subcutaneously injected in sheep and Hanwoo. The serum and meat juice of the immunized or non-immunized animal were then analyzed, to measure the titer of antibody by ELISA and Western blot. The amount of antibodies against the BIO-TAG increased (p<0.05) in serum by vaccination. Furthermore, meat juice from the immunized Hanwoo showed greater (p<0.05) antibody titer, compared with those from non-immunized groups. To optimze the dilution factor, we performed dot-ELISA, with various combination levels of BIO-TAG. Results from dot-ELISA showed that 2 mg/mL BIO-TAG was sufficient to distinguish the immunized meat from non-immunized groups. These results support our hypothesis that simple immunization of Hanwoo generates a sufficient amount of antibodies to be detectable in the meat juice by means of the immune-assay. Therefore, specific Hanwoo brand meat can be more precisely identified by our rapid diagnostic kit. This technology can deter possible fraud of counterfeit meat brands in the Korean domestic market with ease and rapidity; and offers a new tool that guarantees consumers high quality Hanwoo brand beef.

Development of ELISA for Brucella abortus RB51 II. Purification of 8kDa antigen and development of ELISA using its antigen of Brucella abortus RB51 (부루세라 RB51의 ELISA 진단법 개발 II. Brucella abortus RB51균의 8kDa 항원 정제 및 ELISA 진단법 개발)

  • Her, Moon;Cho, Dong-hee;Jung, Byeong-yeal;Cho, Seong-kun;Jung, Suk-chan;Kim, Ok-kyung
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.51-57
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    • 2001
  • A procedure for extraction and purification of 8 kDa antigen of Brucella abortus RB51 was developed. Bacteria heat inactivated at $60^{\circ}C$, 30 min was extracted by 1% sarcosine and followed by fluid pressure liquid gel filtration chromatography of 2 series, Superose 12 HR 10/30 and Sephacryl S-100. There was produced $71.46{\mu}g/g$(wet) of 8 kDa antigen, and it resisted 1% trypsin, solved 1% triton X-100 higher than distilled water and inactivated 0.1% proteinase K. These results show that 8 kDa antigen may be a lipoprotein existed cell surface of B. abortus RB51. Also, we developed ELISA using purified 8 kDa surface antigen of Brucella abortus RB51 strain, its specificity and sensitivity was 95.0%, 98.6%, respectively. As compared with dot-blot assay using whole cell and ELISA using 8 kDa antigen, its correlation was 93.5%.

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