• 제목/요약/키워드: cELISA

검색결과 678건 처리시간 0.029초

RT-PCR 방법을 이용한 효과적인 감자 잎말림 바이러스의 검정 (An Effective Method of Diagnosis of Potato Leafroll Virus by RT-PCR)

  • 전재흥;정영희;최경화;김현순;오현우;박세원;정혁
    • 한국식물병리학회지
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    • 제12권3호
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    • pp.358-362
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    • 1996
  • 감자 잎말림 바이러스를 검정하기 위하여 ELISA 및 전자현미경에 의해 바이러스 감염이 확인된 기내 배양중인 감장의 줄기로부터 RT-PCR 분석을 수행하였다. 분리된 총 RNA들로부터 바이러스 cDNA를 합성하고 감자 잎말림 바이러스 외피단백질의 일부인 465bp를 특이하게 증폭하도록 고안한 두 primer를 사용하여 PCR 반응을 하였다. 증폭된 465pb의 DNA 절편의 염기서열을 분석한 결과 역시 감자 잎말림 바이러스임을 확인하였다. 바이러스 검정에 있어서 EL-ISA 방법과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법간의 민감도를 조사한 결과 RT-PCR 방법이 ELISA 방법보다 감자 잎말림 바이러스검정에 있어서보다 정확한 방법인 것으로 사료된다.

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An Outbreak of Trichinosis with Molecular Identification of Trichinella sp. in Vietnam

  • Nguyen, Van De;Nguyen, Vu Trung;Nguyen, Hong Ha;Vu, Thi Nga;Nguyen, Minh Ha;Pham, Thanh Thuy;Le, Van Duyet;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • 제50권4호
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    • pp.339-343
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    • 2012
  • The 5th outbreak of trichinosis occurred in a mountainous area of North Vietnam in 2012, involving 24 patients among 27 people who consumed raw pork together. Six of these patients visited several hospitals in Hanoi for treatment. Similar clinical symptoms appeared in these patients within 5-8 days after eating infected raw pork, which consisted of fever, muscle pain, difficult moving, edema, difficult swallowing, and difficult breathing. ELISA revealed all (6/6) positive reactions against Trichinella spiralis antigen and all cases showed positive biopsy results for Trichinella sp. larvae in the muscle. The larvae detected in the patients were identified as T. spiralis (Vietnamese strain) by the molecular analysis of the mitochondrial cytochrome c oxidase subunit III (cox3) gene.

전북 동부지역 한우에서의 유사산 유발 질병 항체가 조사 (Seroprevalence of abortion and stillbirth inducing disease in Hanwoo, in Jeonbuk eastern area)

  • 송지민;손구례;고원석;이정원
    • 한국동물위생학회지
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    • 제37권3호
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    • pp.179-183
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    • 2014
  • Reproductive losses in a herd can be huge. Fetal reabsorption or undefined infertility often remain undetected. Routine herds monitoring for exposure, controlling the introduction of potential agent carriers, appropriate biosafety procedures, and vaccination where possible are together the best security against abortion and stillbirth inducing disease. For biosecurity of local farms, we performed antibody titers of abortion and stillbirth related diseases such as bovine viral diarrhea virus (BVDV), infectious bovine rhinotracheitis virus (IBRV), Neospora caninum, Toxoplasma gondii and Campylobacter fetus subsp venerealis. The blood samples were collected from 500 female Hanwoo over 1 year old of 100 farms in Jeonbuk eastern area. Champhylobater serological test was evaluated by the standard tube agglutination test (STAT) and other pathogen's antibodies were detected by indirect-enzyme linked immunoassay (I-ELISA). The seroprevalence of abortion and stillbirth inducing disease were BVDV 72.4%, IBRV 13.0%, N. caninum 1.2%, T. gondii 10.4% and C. venerealis 0.6%, irrespectively.

Production and characterization of monoclonal antibodies (MAb) against flounder serum immunoglobulin (Ig)

  • Jang, Han-Na;Cho, Young-Hye;Park, Sang-Hoon
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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    • pp.446-446
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    • 2000
  • Specific polyclonal and/or monoclonal antibodies (MAbs) to immunoglobulins (Igs) and their subunits have proved to be valuable tools in immunological research and in immunological assays. In this study, we developed and characterized MAbs against flounder serum Igs. To obtain the pure flounder serum Igs, mouse IgG (mIgG) was immunized to flounder. Flounder Igs were purified by using mIgG-agarose affinity column chromatography. The structure of purified flounder Ig was observed, on denatured SDS-PAGE, to be composed of two heavy chains (77 and 72 kd) and two light chains (28 and 26 kd). MAbs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously primed with the flounder Igs. Finally, three hybridoma clones, FIM 511, FIM 519 and FIM 562 were established to recognize both 2 heavy chains, 26 kd of light chain and 28 kd of light chain, respectively. On the other hand, the flounder immune sera collected on the weekly basis were tested on ELISA and immunoblot analysis whether boosting effect is present in flounder humoral immune system. As a result, the secondary immune response in flounder was ascertained on ELISA, but not on immunoblot analysis. Further, we observed an alteration of serum protein levels following immunization. Our MAbs and basic information on flounder humoral immune system obtained in this study will be helpful to control and monitor the efficiency of fish vaccines and therapeutic process of flounder diseases.

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돼지 옴 감염증 진단을 위한 바이오센서 연구 (A Biosensor for Diagnosis of Swine Sarcoptic Mange)

  • 조한근;지차호
    • Journal of Biosystems Engineering
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    • 제30권5호
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    • pp.306-311
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    • 2005
  • In this study, a biosensor was developed to rapidly diagnose the swine sarcoptic mange (Sarcoptes scabies var. suis). The ELISA was modified to reduce the processing time for rapid diagnosis. The biosensor consists of a biological reaction part, and a measurement and control part. The biological reaction part was designed for using micro-pumps and valves for fluid transportation, and the measurement control part composed of a photodiode, a light-emitting diode fur light measurement, and a microcomputer to implement assay A polystyrene covet was used as a reaction chamber. Signal output was read as the rate of change in optical density at 645nm. Eighteen pigs diagnosed with sacroptic mange and 19 control pigs were tested. Fifteen sacbies-infested pigs showed positive results ($83.3\%$ sensitivity). Sixteen control pigs showed negative results ($84.2\%$ specificity). The system could execute a diagnosis cycle in about 45 min. The results suggest that this biosensor is useful for the rapid diagnosis of swine sacroptic mange.

Tissue parasitic helminthiases are prevalent at Cheongjin, North Korea

  • Shen, Chenghua;Li, Shunyu;Zheng, Shanzi;Choi, Min-Ho;Bae, Young-Mee;Hong, Sung-Tae
    • Parasites, Hosts and Diseases
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    • 제45권2호
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    • pp.139-144
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    • 2007
  • We investigated a small-scale serological survey to screen tissue-parasitic helminthiases of North Koreans as one of research programs for re-unification of Korea. Soil-transmitted helminthiases were found highly prevalent among North Korean residents at the border with China. ELISA using 4 tissue-parasitic helminth antigens was applied to 137 residents living in Cheongjin-shi, Hamgyeongbuk-do, North Korea and 133 female refugees in South Korea in 2004-2005. Among a total of 270 samples, 31 (11.5%), 25 (9.3%), and 11 (4.1%) were positive for specific IgG antibodies to antigens of Clonorchis sinensis, Taenia solium metacestode, and sparganum, respectively. The overall positive rate was 21.5%; 38.2% in males and 15.8% in females. The present finding suggests that tissue parasites, such as C. sinensis, T. solium metacestode and sparganum are highly prevalent in some limited areas of North Korea. These foodborne tissue-parasitic helminthiases should be considered for future control measures of parasitic diseases in North Korea.

효소가수분해에 의한 유청단백질의 항원성 저하 (Reduction of the Antigenicity of Whey Protein by Enzymatic Hydrolysis)

  • 하월규;전석락;김정완;이수원;이재영;손동화
    • 한국식품과학회지
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    • 제26권1호
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    • pp.74-80
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    • 1994
  • 효소에 의한 단백질분해가 유청단백질의 항원성의 저하에 미치는 영향을 조사하기 위한 기본연구로서, 유청단백질의 가수분해특성을 조사하고 competitive inhibition enzyme-linked immunosorbent assay(cELISA)에 의한 항원성의 변화를 검토하였다. 유청단백질의 가수분해는 chymotrypsin, trypsin, pancreatin, 그리고 Aspergillus oryzae유래의 protease를 각기 4시간 동안 행하였다. TNBS(trinitrobenzensulfonic acid)법에 의하여 측정한 유청단백질의 가수분해도(DH)는 chymotrypsin이나 trypsin을 처리한 경우$(5.05{\sim}11.47)$보다 Aspergillus oryzae유래의 protease 및 pancreatin을 처리한 경우$(15.67{\sim}20.20)$가 훨씬 높게 나타났으며, 각 효소의 처리전에 열처리($75^{\circ}C$, 20분)나 pepsin의 처리를 한 경우에 대체로 약간 높게 나타났다. High performance size exclusion chromatography(HPSEC)에 의하여 분자량분포를 조사한 결과, 가수분해물에 따라 10kDa 이상의 polypeptide가 $12{\sim}36%$ 정도 존재하였고, 평균분자량은 $4,252{\sim}9,132$ dalton, 평균길이는 아미노산 $38{\sim}83$개로 나타났다. 또한 쓴맛은 형성되지 않았다. SDS-PAGE의 결과 처리구에 따라 분자량 14.2kDa 이상의 polypeptide가 일부 존재하였으나 native 유청단백질은 대부분 가수분해에 의하여 제거되었음을 확인하였다. 토끼 항WPI항혈청에 의한 cELISA로 검토한 유청단백질 가수분해물의 monovalent 항원성은 효소처리에 의하여 약 $10^{-1.7}{\sim}10^{-4.9}$배 또는 그 이하로 저하되었으며 대체로 가수분해가 많이 일어난 분해물은 그 항원성이 낮아지는 것으로 나타났다. 또한 각 처리구내에서는 열 및 pepsin의 전처리후 다음 효소 분해한 유청단백질 가수분해물(CDP, TDP, PDP, ODP)의 경우 그 항원성이 가장 낮았다. 그중에서도 pancreatin 가수분해물(PDP)의 경우 항원성이 거의 상실된 것으로 나타났다.

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지골피(地骨皮)가 강제수영부하실험에서 HPA-Axis System에 미치는 영향 (Effects of Lycii Radicis Cortex in HPA-Axis System on the Forced Swimming Test)

  • 구범모;이태희
    • 대한한의학방제학회지
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    • 제18권2호
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    • pp.215-225
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    • 2010
  • In order to investigate the antidepressant effects of Lycii Radicis Cortex on the change of HPA-Axis system, the forced swimming test was performed. The expressions of CRF and c-Fos in the PVN were measured with immunohistochemical method and the concentration of ACTH in Serum was evaluated with ELISA method. And the results obtained were as follows. Results : 1. The duration of immobility in the forced swimming test was significantly decreased in the LRC100 group and the LRC400 group(P<0.001). 2. The expressions of CRF and c-Fos were significantly reduced in the LRC100 group and the LRC400 group(P<0.001). And the concentration of ACTH in Serum were significantly reduced in the LRC 100 group(P<0.05). According to the results above mentioned, it can be considered that Lycii Radicis Cortex has antidepressant effects.

캔디다질염에 효과가 있는 단항체에 대한 호중구의 역할 (A Role of Neutrophils in Anti-Candida Monoclonal Antibody Protection Against Vaginal Infection due to Candida albicans)

  • 한용문
    • 약학회지
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    • 제47권3호
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    • pp.190-194
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    • 2003
  • As previously reported, an immunoglobulin M (IgM) monoclonal antibody (MAb) B6.1, specific for a cell wall B-l,2-mannotriose, was protective against vaginal infection due to Candida albicans when mice were treated with the antibody. In this study, the role of neutrophil was examined in the protective effect of MAb B6.1 against vaginal infection. To deplete neutrophils, mice were given intravenously rat anti-mouse neutrophile MAb RB6-8C5 prior to intraperitoneal administration of MAb B6.1 to these mice. The mice were examined for antibody in their reproductive tract. By an ELISA, MAb B6.1 was found in the vaginal homogenates, but no antibody was detected in vaginal lavage materials. The neutropenia was induced by a single dose of the anti-neutrophil antibody, but lymphocytes were also partially depleted. The protective effect of MAb B6.1 was decreased when mice pretreated with MAb RB6-8C5 were given the anti-Candida antibody before challenge with C. albicans yeast cells intravaginally. These results show that neutrophils are involved in the MAb B6.1 protection against Candida vaginal infection.

Preparation and Characterization of Anti-GP73 Monoclonal Antibodies and Development of Double-antibody Sandwich ELISA

  • Li, Qi-Wen;Chen, Hong-Bing;Li, Zhi-Yang;Shen, Peng;Qu, Li-Li;Gong, Lai-Ling;Xu, Hong-Pan;Pang, Lu;Si, Jin
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권5호
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    • pp.2043-2049
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    • 2015
  • Background: Serum Golgi protein 73 (GP73) as a novel and potential marker for diagnosing hepatocellular carcinoma (HCC) have been found to be elevated in HCC patients and associated with clinical variables representing tumor growth and invasiveness. The aim of this study was to prepare a pair of monoclonal antibodys (mAbs) against GP73 and develop a newly designed double-antibody sandwich enzyme-linked immunosorbent assay (s-ELISA), which would be used in the detection of serum GP73 (sGP73) as well as in the diagnosis of HCC. Materials and Methods: Produced by prokaryotic expression, the purified recombinant GP73 (rGP73), produced by prokaryotic expression, was used to immunize the Balb/c mice. Two hybridoma cell lines against GP73 were obtained by fusing mouse Sp2/0 myeloma cells with spleen cells from the immunized mice. The titers of anti-GP73 mAb reached 1:243,000. Western blotting analysis and Immunohistochemistry staining revealed that anti-GP73 mAb could recognize GP73 protein. The double-antibody s-ELISA was successfully established and validated by 119 HCC and 103 normal serum samples. Results: showed that the detection limit of this method could reach 1.56 ng/ml, and sGP73 levels in HCC group (mean=190.6 ng/ml) were much higher than those of in healthy controls (mean=70.92 ng/ml). Conclusions: Results of our study not only showed that sGP73 levels of HCC patients were significantly higher than those of healthy controls, but also indicated that the laboratory homemade anti-GP73 mAbs could be the optimal tool used in evaluating sGP73 levels, which would provide a solid foundation for subsequent clinical applications.