• 대한바이러스학회지
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• 제26권1호
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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.

• 대한수의학회지
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• 제31권4호
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• pp.403-409
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• 1991

Progesterone의 단크론성 항체를 생산, 이용하여 감도가 높으면서도 신속히 측정할 수 있는 ELISA 기법을 처음으로 개발코져 실시하였다. 단크론성 항체는 종래의 면역방법에 의해 획득한 항혈청에 비해 약 10배의 결합율을 보였고 titer 역시 높았다. Dot-blot 분석 결과 단크론성 항체는 IgM이었다. 경합반응은 2시간으로 충분하였고, progesterone 표준용액을 이용한 표준 곡선은 0~1000pg/well에서 거의 직선적이었다. Progesterone의 단크론성 항체를 이용한 ELISA는 임상적으로는 물론 연구용으로도 신속한 항체의 기능 측정에는 물론 각종 번식 관련의 지표로 충분히 활용될 수 있을 것으로 판단된다.

• 대한수의학회지
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• 제41권1호
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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.

• 한국동물학회지
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• 제35권2호
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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.

• The Plant Pathology Journal
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• 제34권2호
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• pp.85-92
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• 2018

Enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR), widely used for the detection of plant viruses, are not easily performed, resulting in a demand for an innovative and more efficient diagnostic method. This paper summarizes the characteristics and research trends of biosensors focusing on the physicochemical properties of both interface elements and bioconjugates. In particular, the topological and photophysical properties of quantum dots (QDs) are discussed, along with QD-based biosensors and their practical applications. The QD-based Fluorescence Resonance Energy Transfer (FRET) genosensor, most widely used in the biomolecule detection fields, and QD-based nanosensor for Rev-RRE interaction assay are presented as examples. In recent years, QD-based biosensors have emerged as a new class of sensor and are expected to open opportunities in plant virus detection, but as yet there have been very few practical applications (Table 3). In this article, the details of those cases and their significance for the future of plant virus detection will be discussed.

• 한국축산식품학회지
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• 제34권3호
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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.

• The Plant Pathology Journal
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• 제36권6호
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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.

• The Plant Pathology Journal
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• 제26권1호
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• pp.25-31
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• 2010

Papaya ringspot virus (PRSV) causes the most widespread and devastating disease in papaya. Isolates of PRSV originating from different geographical regions in south India were collected and maintained on natural host papaya. The entire coat protein (CP) gene of Papaya ringspot virus-P biotype (PRSV-P) was amplified by RTPCR. The amplicon was inserted into pGEM-T vector, sequenced and sub cloned into a bacterial expression vector pRSET-A using a directional cloning strategy. The PRSV coat protein was over-expressed as a fusion protein in Escherichia coli. SDS-PAGE gel revealed that CP expressed as a ~40 kDa protein. The recombinant coat protein (rCP) fused with 6x His-tag was purified from E.coli using Ni-NTA resin. The antigenicity of the fusion protein was determined by western blot analysis using antibodies raised against purified PRSV. The purified rCP was used as an antigen to produce high titer PRSV specific polyclonal antiserum. The resulting antiserum was used to develop an immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) assay and compared its sensitivity levels with ELISA based assays for detection of PRSV isolates. IC-RT-PCR was shown to be the most sensitive test followed by dot-blot immunobinding assay (DBIA) and plate trapped ELISA.

• 한국작물학회지
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• 제44권4호
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• pp.339-344
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• 1999

Immunological method has been applied in biochemical genetic analysis of seed storage proteins. We developed and characterized anti-gliadin polyclonal antibody (AGPab) specific to gliadin fractions whose quality and quantity were known to be associated with wheat end-use quality. Reactions of anti-gliadin polyclonal antibody (AGPab) to gliadin were linearly decreased as AGPab and antigen were diluted. Dot-blot and immunoblot assay showed that produced AGPab specifically reacted to gliadin and mainly $\alpha$-, $\beta$-, and ${\gamma}$-gliadin subunits. Enzyme-linked immuno- sorbent assay (ELISA) was applied for quantifi-cation of gliadins in Korean wheat cultivars and breeding lines by using AGPab. High reactions between AGPab and gliadins were found in wheat cultivars Olmil and Olgeurumil. Significant difference of optical densities for alcohol soluble proteins among crop species was found, as wheat showed the highest value (0.697) followed by rye (0.295), and barley (0.066).

• 대한수의학회지
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• 제41권1호
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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%.