• Title/Summary/Keyword: Colloidal gold

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The Localization of Lectin Receptors in the Tissue of the Paragonimus westermani (폐흡충조직내 Lectin(WGA) 수용체의 분포)

  • Kim, Soo-Jin;Nahm, Heun-Woo;Lee, Joon-Sang;Joo, Kyung-Whan
    • Applied Microscopy
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    • v.30 no.1
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    • pp.101-111
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    • 2000
  • In this study, the distribution of lectin receptors in Paragonimus westermani tissue was explored using colloidal gold label complexed with lectin WGA purified from wheat germ (Triticum vulgare). The lectin WGA gold complex, shown to recognize GlcNAc (N-acetylgalactosamine) and NeuNAc (N-acetylneuraminic acid) regions, was applied to detect binding sites in Lowicryl HM 20 sections viewed under electron microscope. Labeled sections of the metacercaria revealed gold particles specifically distributed on the tegumental syncytium and lamella of the excretory canal. Labeling of young adult tissue was then quantified and compared to that of adult worm tissue. Adult worm tissue sections resulted in specific gold particle distribution on the lamella of caecal epithelium and excretory canal. These results indicate that lectin WGA receptors are located in the tegumental syncytium and lamella of the excretory canal of the metacercariae, and in the lamella of the caecum and excretory canal of the young adult and adult. Therefore, the GlcNAc and NeuNAc regions in the tegumental syncytium appear to be functionally associated with cell-recognition and protection from the immune system of the host, and linked with membrane transport and absorption of nutrients in the lamella of the excretaory canal and caecal epithelia.

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Development of One-step Simultaneous Immunochromatographic Assay for Rapid Analysis of Aflatoxin B1 and Ochratoxin A

  • Shim, Won-Bo;Kim, Gyeong-Yeol;Ryu, Hee-Jung;Nam, Min-Ji;Chung, Duck-Hwa
    • Food Science and Biotechnology
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    • v.18 no.3
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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.

Development of Immunochromatography for Detection of Escherichia coli O157:H7 Contaminated in Meats and Sprouts (육류 및 새싹채소에 오염된 Escherichia coli O157:H7의 신속검출을 위한 면역크로마토그래피법의 개발)

  • Lee, Won-Gyeong;Ryu, Hee-Jeong;Song, Jeong-Un;Kim, Kyeong-Yeol;Chung, Do-Yeong;Kim, Jeong-Sook;Shim, Won-Bo;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.43 no.5
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    • pp.618-623
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    • 2011
  • Escherichia coli O157:H7 has been considered as a significant food-borne pathogen since its role in causing hemorrhagic colitis and hemolytic uremic syndrome in humans was recognized. In this study, we developed an immunochromatography (ICG) assay for the detection of E. coli O157:H7. E. coli O157:H7 monoclonal antibody (EC MAb) and colloidal gold were conjugated and its specificity was determined by the ICG treated with EC MAb and antimouse IgG at test and control lines, respectively. The detection limit of the ICG was $1{\times}10^5$ CFU/mL, and no crossreactivity was observed to other E. coli strains and major food-borne pathogens. To determine the minimum enrichment time for the ICG, meats and sprouts were inoculated with $1{\times}10$ CFU/100 ${\mu}L$ of E. coli O157:H7. After enrichment time of 10 and 2 h for meats and sprouts, respectively, up to $1{\times}10$ CFU/100 ${\mu}L$ of E. coli O157:H7 could be detected by ICG.

Rapid Detection Kit for Salmonella typhimurium (살모넬라 식중독균 신속 검출용 간이 진단키트)

  • Kim, Gi-Young;Yang, Gil-Mo;Park, Saet-Byeol;Kim, Yung-Hwun;Lee, Kang-Jin;Son, Jae-Yong;Kim, Hyuck-Joo;Lee, Sae-Rom
    • Journal of Biosystems Engineering
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    • v.36 no.2
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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.

Rapid Detection Kit for Staphylococcus aureus (황색포도상구균 신속 검출용 간이 진단키트)

  • Kim, Gi-Young;Park, Saet-Byeol;Moon, Ji-Hea;Lee, Sang-Dae;Lee, Sea-Rom;Jang, Youn-Jung
    • Korean Journal of Agricultural Science
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    • v.40 no.2
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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.

An Immuno-Electronmicroscopic Study on the Synoviocytes in the Knee Joint of the Human (인체 무릎관절 윤활세포에 관한 면역전자현미경적 연구)

  • Hwang, Douk-Ho;Chang, Ka-Young;Lee, Wang-Jae;Park, Kyung-Han;Lee, Jong-Bum
    • Applied Microscopy
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    • v.26 no.1
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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.

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Synthesis and Characterization of Photosensitizer-conjugated Gold Nanorods for Photodynamic/Photothermal Therapy (광역학적/광열치료 응용을 위한 광증감제가 결합한 골드 나노로드의 합성 및 특성분석)

  • Choi, Jongseon;Kim, So Yeon
    • Applied Chemistry for Engineering
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    • v.27 no.6
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    • pp.599-605
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    • 2016
  • Recently, photodynamic and photothermal therapies have received increasing attention as an effective cancer treatment. In this study, a gold nanorod (AuNR) colloidal solution was synthesized as a hyperthermia agent for photothermal therapy and also modified with photosensitizer (PS) for photodynamic therapy. PEG (polyethylene glycol) and FA (folic acid) ligand were also introduced into AuNR for the long circulation in human body and efficient targeting of cancer cells, respectively and AuNRs were modified with FA-PEG and poly-${\beta}$-benzyl-L-aspartate (PBLA) block copolymers through a 3,4-dihydroxy hydrocinnamic acid (HCA) linker. A series of AuNRs with various aspect ratios were synthesized by controlling the feeding ratio of $AgNO_3$. The physicochemical property and morphology of synthesized AuNR100 and FA-PEG-$P(Asp)_{50}$-HCA-AuNR100 were analyzed by UV-visible spectrophotometer, $^1H$ NMR, XPS measurements, TEM. The surface modified AuNR carrier with biocompatibility could be applied for the effective diagnosis as well tumor phototherapy.

Preparation and Characterization of Thiolate-Protected Gold Nanoparticles Using Modified One-Phase Method (개선된 단일상 합성법을 이용한 티올화 나노 금의 합성 및 확인)

  • Park, Jisu;Kim, Youhyuk
    • Journal of the Korean Chemical Society
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    • v.61 no.4
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    • pp.191-196
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    • 2017
  • One-phase method to prevent the initial formation of Ag(I) thiolate layered materal from the mixture of $AgNO_3$ and thiols was previously developed to generate TP (Thiolate-Protected)-nanosilver. In this modified method, $AgNO_3$ is added to the mixtures of $NaBH_4$ and thiols in ethanol. This method was so successful that it was applied to synthesize TP-nanogold and nanoplatinum. The synthesis and characterization of these nanoparticles by ultraviolet-visible (UV-vis) absorption spectra, transmission electron microscopy (TEM) pictures, X-ray powder differaction (XRD) patterns and infrared(IR) spectra are described. The results show that colloidal nanoparticles are spherical or oval shape and the mean sizes for TP-nanogold and nanoplatinum are about 3~7 nm and below 2 nm, respectively. The conformation of polymethylene [$-(CH_2)_7-$] sequence in octanethiolate attached to nanogold was elucidated as trans.

Development of a Rapid Diagnostic Test Kit to Detect IgG/IgM Antibody against Zika Virus Using Monoclonal Antibodies to the Envelope and Non-structural Protein 1 of the Virus

  • Kim, Yeong Hoon;Lee, Jihoo;Kim, Young-Eun;Chong, Chom-Kyu;Pinchemel, Yanaihara;Reisdorfer, Francis;Coelho, Joyce Brito;Dias, Ronaldo Ferreira;Bae, Pan Kee;Gusmao, Zuinara Pereira Maia;Ahn, Hye-Jin;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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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.

White Sport Syndrome Virus Disease of Shirmp and Diagnostic Methods

  • Zhan, Wen-Bin
    • Journal of Aquaculture
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    • v.15 no.1
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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.