• Applied Microscopy
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• 제30권1호
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• pp.101-111
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• 2000

기생충에서 세포 표면에 존재하는 당단백 말단 Glc-NAc(N-acetylglucosa-mine)와 NeuNAc (N-acetylneuraminic acid)가 숙주 면역계를 인식하고 영양물질 흡수와 막 투과성에 중요한 역할을 하는 것으로 알려졌다. 이들 당단백 말단 GlcNAc와 NeuNAc의 폐흡충 피낭유충과 유약성충, 성충의 충체 조직세포에 분포를 확인하기 위하여 WGA 황금입자 복합체를 반응시켰다. WGA 황금입자 복합체에 반응시킨 폐흡충 발육 단계별 조직을 전자현미경으로 관찰한 결과 폐흡충 피낭유충의 표피합포체에는 황금입자가 고밀도로 표지된 것이 관찰되어 폐흡충 피낭유충 표피세포질에 lectin수용체들이 고밀도로 분포하는 것으로 관찰되었다. 유약성충과 성충의 맹관 막구조물에 WGA 황금입자가 고밀도로 표지되어 맹관의 상피세포 막 구조물에 WGA 수용체들이 고밀도로 분포하는 것으로 확인되었다. 폐흡충 피낭유충과 유약성충 그리고 성충의 배설관 상피의 막구조물에 WGA 황금입자가 표지되어 배설관상피의 막구조물에 WGA 수용체가 분포하는 것으로 확인되었다. 따라서 피낭유충의 표피합포체와 표피세포의 세포질에 분포하는 WGA 수용체인 GlcNAc와 NeuNAc는 숙주의 면역계와 세포인식에 관여하며 유약성충과 성충은 표피세포의 GlcNAc와 NeuNAc는 소멸되고 맹관과 배설관 상피에 GlcNAc와 NeuNAc가 분포하여 영양물질 흡수와 막수송에 의한 재흡수작용이 활발한 것이 확인되었다.

• Food Science and Biotechnology
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• 제18권3호
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• pp.641-648
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• 2009

A one-step simultaneous immunchromatographic (OS-ICG) assay using colloidal gold-monoclonal antibody (gold-MAb) conjugates was developed for the rapid multianalysis of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in feed samples. Visual detection limits for AFB1 and OTA were 0.5 and 2.5 ng/mL, respectively, and the results were obtained within 15 min. Matrix interference from the feed extracts was efficiently reduced by appropriate dilution with buffer. Cut-off values of the OS-ICG assay for the feed spiked with AFB1/OTA mixtures (5/5, 10/10, 25/25, 50/55, 100/100 ${\mu}g/kg$) were 10 and 50 ${\mu}g/kg$ for AFB1 and OTA. The comparative analyses of 65 feed samples by OS-ICG, enzyme-linked immunosorbent assay (ELISA), and high performance liquid chromatography (HPLC) showed good agreement. In this study, we confirmed that simultaneous analysis based on immunoassay is possible and it can be used as an on-site multianalysis of AFB1 and OTA in feed, food, and agricultural products.

• 한국식품과학회지
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• 제43권5호
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• pp.618-623
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• 2011

E. coli O157:H7은 요독증후군, 출혈성 대장염 및 설사 등을 유발하는 원인균으로, 전세계적으로 이들의 오염에 따른 식중독 발생이 증가하고 있어, 본 연구에서는 E. coli O157:H7을 신속 정확하게 분석할 수 있는 면역크로마토그래피법을 개발하였고, 보다 신속한 검출을 위하여 육류와 새싹시료를 대상으로 최소 증균시간을 확인하였다. 먼저, 면역크로마토그래피법의 개발을 위해 colloidal gold와 EC MAb를 합성하여 EC MAb-gold conjugate를 제작한 다음, 접합여부를 확인한 결과, test line과 control line에 각각 처리되는 EC MAb와 anti-mouse IgG에 특이적으로 반응하였다. 제작된 EC MAb-gold conjugate를 이용하여 개발된 면역크로마토그래피법의 검출한계는 $1{\times}10^5$ CFU/mL 수준까지 검출이 가능한 것으로 확인되었고, 다른 대장균속 및 주요 식중독균과의 교차반응성은 나타나지 않았다. 개발된 면역크로마토그래피법을 이용하여 임의로 E. coli O157:H7을 오염시킨 돼지고기, 소고기, 닭고기 및 새싹채소를 증균하는 동안 분석한 결과, 돼지고기와 닭고기는 6시간, 소고기는 10시간, 특히 새싹채소는 2시간 후부터 $1{\times}10$ CFU/100 ${\mu}L$ 수준까지 검출이 가능한 것으로 나타났다. 따라서 본 연구에서 개발된 면역크로마토그래피법을 이용하여 육류와 채소 등에 오염된 E. coli O157:H7을 최대 10시간 이내로 신속히 분석할 수 있을 것으로 판단된다.

• Journal of Biosystems Engineering
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• 제36권2호
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• pp.140-146
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• 2011

This study was performed to develop a rapid test kit for pathogenic Salmonella in various samples. The rapid detection kit has been fabricated based on nitrocellulose lateral-flow strip. Colloidal gold and biotin conjugated Salmonella antibodies were used as a tag and a receptor, respectively. Manually spotted Salmonella antibody and Neutravidin on nitrocellulose membrane were used as test and control lines, respectively. Feasibility of the rapid kit to detect Salmonella typhimurium in samples were evaluated. The intensity of the color of the test line started to increase with the samples in which higher concentration of the cells were contained. The sensitivity of the sensor was $10^6$ cfu/mL Salmonella spiked in PBS. Also, the rapid test kit could detect $10^6$ cfu/mL of Salmonella in chicken meat extract.

• 농업과학연구
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• 제40권2호
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• pp.139-146
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• 2013

This study was performed to develop a rapid test kit for pathogenic Staphylococcus in various samples. The rapid detection kit has been fabricated based on nitrocellulose lateral-flow strip. Colloidal gold and Staphylococcus antibodies were used as a tag and a receptor, respectively. Manually spotted Staphylococcus antibody and anti-mouse antibody on the surface of nitrocellulose membrane were used as test and control lines, respectively. Feasibility of the rapid kit to detect Staphylococcus aureus in samples were evaluated. The intensity of the color of the test line started to increase with the samples in which higher concentration of the cells were contained. The sensitivity of the sensor was $10^6$ cfu/mL Staphylococcus spiked in PBS. Also, the rapid test kit could detect $10^5$ cfu/mL of Staphylococcus in chicken meat extract.

• Applied Microscopy
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• 제26권1호
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• pp.11-16
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• 1996

This study was designed to observe the ultrastructure of synoviocytes which are concerned with phagocytic function in the knee joint of the human. The synovia were dissected and were fixed for two hours in 0.2% glutaraldehyde and 4% paraformaldehyde solution and processed and finally infused in 2.3 M sucrose and 20% PVP solution. The tissues were cut with the cryoultramicrotome and labelled with primary antibodies (anti-tubulin, anti-vimentin) and secondary antibody-6 nm colloidal gold particles. The tissues were observed under transmission electronmicroscope. The results were followings. 1. In phagocytic synovial cells, the distributions of tubulin were cytoplasm, especially around vacuoles. 2. In phagocytic synovial cells, the distributions of vimentin were cytoplasm. 3. Both tubulin and vimentin were not located inside of vacuoles. On the basis of above findings, it is obvious that the phagocytic functions are concerned with tubulin, and the phagocytic synovial cells contain vimentin.

• 공업화학
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• 제27권6호
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• pp.599-605
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• 2016

최근 효과적인 암 치료 방법으로 광역학치료(photodynamic therapy)와 광열치료(photothermal therapy)가 주목받고 있다. 본 연구에서는 광열치료에 필요한 광열인자로써의 역할을 할 수 있는 골드 나노로드(AuNR)를 합성하고, 그 표면에 광역학치료를 위한 광증감제(photosensitizer)를 결합하였다. 즉, 골드 나노로드를 체내에 오래 머무르도록 하기 위해 PEG(polyethylene glycol) 및 효과적인 암 표적지향성을 위해 FA (folic acid) 리간드를 도입하였고, FA-PEG와 poly-${\beta}$-benzyl-L-aspartate (PBLA)로 이루어진 블록 공중합체를 3,4-dihydroxy hydrocinnamic acid (HCA) linker를 사용하여 골드 나노로드의 표면개질을 하였다. 또한 $AgNO_3$의 feeding ratio 변화를 통해 다양한 aspect ratio를 갖는 골드 나노로드를 합성하였고, UV-visible spectrophotometer, $^1H$-NMR, XPS, TEM 분석을 통해 FA-PEG-$P(Asp)_{50}$-HCA-AuNR100의 물리 화학적 특성과 morphology를 분석하였고, 성공적인 표면 개질을 확인할 수 있었다. 골드 나노로드의 표면 개질을 통한 생체 적합성 약물전달체의 합성은 효과적인 암 진단 및 다양한 광역학/광열치료 분야에 응용이 될 수 있을 것으로 기대된다.

• 대한화학회지
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• 제61권4호
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• pp.191-196
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• 2017

앞선 연구에서 TP(Thiolate-Protected)-나노은의 합성 시 초기에 형성되는 티올화 은 층간화합물의 형성을 막기 위하여 에탄올에 녹아있는 $NaBH_4$와 티올에 $AgNO_3$를 첨가하는 개선된 단일상 합성법을 보고하였다. 본 연구에서는 이 합성법을 보다 일반화하기 위하여 TP-나노금과 나노백금의 합성에 적용하였다. 합성된 생성물의 나노크기, 형상 및 금속에 부착된 티올의 배열은 UV-vis. 스펙트럼, 투과 전자 현미경 사진, X-선 회절 패턴 및 적외선 분광 스펙트럼을 이용하여 규명 하였다. TP-나노금과 나노백금은 구형 및 타원형의 형태를 하고 있으며 입자크기는 각각 약 3~7 nm와 약 2 nm 이하로 나타나고 있다. 한편 TP-나노금의 표면을 감싸는 옥탄티올 음이온의 메틸렌 사슬 [$-(CH_2)_7-$]의 메틸렌 배향은 trans임을 보여주고 있다.

• Parasites, Hosts and Diseases
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• 제56권1호
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• pp.61-70
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• 2018

We developed a Rapid Diagnostic Test (RDT) kit for detecting IgG/IgM antibodies against Zika virus (ZIKV) using monoclonal antibodies to the envelope (E) and non-structural protein 1 (NS1) of ZIKV. These proteins were produced using baculovirus expression vector with Sf9 cells. Monoclonal antibodies J2G7 to NS1 and J5E1 to E protein were selected and conjugated with colloidal gold to produce the Zika IgG/IgM RDT kit (Zika RDT). Comparisons with ELISA, plaque reduction neutralization test (PRNT), and PCR were done to investigate the analytical sensitivity of Zika RDT, which resulted in 100% identical results. Sensitivity and specificity of Zika RDT in a field test was determined using positive and negative samples from Brazil and Korea. The diagnostic accuracy of Zika RDT was fairly high; sensitivity and specificity for IgG was 99.0 and 99.3%, respectively, while for IgM it was 96.7 and 98.7%, respectively. Cross reaction with dengue virus was evaluated using anti-Dengue Mixed Titer Performance Panel (PVD201), in which the Zika RDT showed cross-reactions with DENV in 16.7% and 5.6% in IgG and IgM, respectively. Cross reactions were not observed with West Nile, yellow fever, and hepatitis C virus infected sera. Zika RDT kit is very simple to use, rapid to assay, and very sensitive, and highly specific. Therefore, it would serve as a choice of method for point-of-care diagnosis and large scale surveys of ZIKV infection under clinical or field conditions worldwide in endemic areas.

• 한국양식학회지
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• 제15권1호
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• pp.7-13
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• 2002

Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.