• Title/Summary/Keyword: Molecular Detection

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Coating gold nanoparticles to a glass substrate by spin-coat method as a surface-enhanced raman spectroscopy (SERS) plasmonic sensor to detect molecular vibrations of bisphenol-a (BPA)

  • Eskandari, Vahid;Hadi, Amin;Sahbafar, Hossein
    • Advances in nano research
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    • v.13 no.5
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    • pp.417-426
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    • 2022
  • Bisphenol A (BPA) is one of the chemicals used in monomer epoxy resins and polycarbonate plastics. The surface-enhanced Raman spectroscopy (SERS) method is precise for identifying biological materials and chemicals at considerably low concentrations. In the present article, the substrates coated with gold nanoparticles have been studied to identify BPA and control the diseases caused by this chemical. Gold nanoparticles were made by a simple chemical method and by applying gold salt and trisodium citrate dihydrate reductant and were coated on glass substrates by a spin-coat approach. Finally, using these SERS substrates as plasmonic sensors and Raman spectroscopy, the Raman signal enhancement of molecular vibrations of BPA was investigated. Then, the molecular vibrations of BPA in some consumer goods were identified by applying SERS substrates as plasmonic sensors and Raman spectroscopy. The fabricated gold nanoparticles are spherical and quasi-spherical nanoparticles that confirm the formation of gold nanoparticles by observing the plasmon resonance peak at 517 nm. Active SERS substrates have been coated with nanoparticles, which improve the Raman signal. The enhancement of the Raman signal is due to the resonance of the surface plasmons of the nanoparticles. Active SERS substrates, gold nanoparticles deposited on a glass substrate, were fabricated for the detection of BPA; a detection limit of 10-9 M and a relative standard deviation (RSD) equal to 4.17% were obtained for ten repeated measurements in the concentration of 10-9 M. Hence, the Raman results indicate that the active SERS substrates, gold nanoparticles for the detection of BPA along with the developed methods, show promising results for SERS-based studies and can lead to the development of microsensors. In Raman spectroscopy, SERS active substrate coated with gold nanoparticles are of interest, which is larger than gold particles due to the resonance of the surface plasmons of gold nanoparticles and the scattering of light from gold particles since the Raman signal amplifies the molecular vibrations of BPA. By decreasing the concentration of BPA deposited on the active SERS substrates, the Raman signal is also weakened due to the reduction of molecular vibrations. By increasing the surface roughness of the active SERS substrates, the Raman signal can be enhanced due to increased light scattering from rough centers, which are the same as the larger particles created throughout the deposition by the spin-coat method, and as a result, they enhance the signal by increasing the scattering of light. Then, the molecular vibrations of BPA were identified in some consumer goods by SERS substrates as plasmonic sensors and Raman spectroscopy.

Rapidly quantitative detection of Nosema ceranae in honeybees using ultra-rapid real-time quantitative PCR

  • Truong, A-Tai;Sevin, Sedat;Kim, Seonmi;Yoo, Mi-Sun;Cho, Yun Sang;Yoon, Byoungsu
    • Journal of Veterinary Science
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    • v.22 no.3
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    • pp.40.1-40.12
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    • 2021
  • Background: The microsporidian parasite Nosema ceranae is a global problem in honeybee populations and is known to cause winter mortality. A sensitive and rapid tool for stable quantitative detection is necessary to establish further research related to the diagnosis, prevention, and treatment of this pathogen. Objectives: The present study aimed to develop a quantitative method that incorporates ultra-rapid real-time quantitative polymerase chain reaction (UR-qPCR) for the rapid enumeration of N. ceranae in infected bees. Methods: A procedure for UR-qPCR detection of N. ceranae was developed, and the advantages of molecular detection were evaluated in comparison with microscopic enumeration. Results: UR-qPCR was more sensitive than microscopic enumeration for detecting two copies of N. ceranae DNA and 24 spores per bee. Meanwhile, the limit of detection by microscopy was 2.40 × 104 spores/bee, and the stable detection level was ≥ 2.40 × 105 spores/bee. The results of N. ceranae calculations from the infected honeybees and purified spores by UR-qPCR showed that the DNA copy number was approximately 8-fold higher than the spore count. Additionally, honeybees infected with N. ceranae with 2.74 × 104 copies of N. ceranae DNA were incapable of detection by microscopy. The results of quantitative analysis using UR-qPCR were accomplished within 20 min. Conclusions: UR-qPCR is expected to be the most rapid molecular method for Nosema detection and has been developed for diagnosing nosemosis at low levels of infection.

Alternative and Rapid Detection Methods for Wastewater Surveillance of SARS-CoV-2 (SARS-CoV-2의 하수조사를 위한 대체 및 신속 검출 방법)

  • Jesmin Akter;Bokjin Lee;Jai-Yeop Lee;Chang Hyuk Ahn;Nishimura Fumitake;ILHO KIM
    • Journal of Korean Society on Water Environment
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    • v.40 no.1
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    • pp.19-35
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    • 2024
  • The global pandemic, coronavirus disease caused by Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to the implementation of wastewater surveillance as a means to monitor the spread of SARS-CoV-2 prevalence in the community. The challenging aspect of establishing wastewater surveillance requires a well-equipped laboratory for wastewater sample analysis. According to previous studies, RT-PCR-based molecular tests are the most widely used and popular detection method worldwide. However, this approach for the detection or quantification of SARS-CoV-2 from wastewater demands a specialized laboratory, skilled personnel, expensive instruments, and a workflow that typically takes 6 to 8 hours to provide results for a few samples. Rapid and reliable alternative detection methods are needed to enable less-well-qualified practitioners to set up and provide sensitive detection of SARS-CoV-2 within wastewater at regional laboratories. In some cases, the structural and molecular characteristics of SARS-CoV-2 are unknown, and various strategies for the correct diagnosis of COVID-19 have been proposed by research laboratories. The ongoing research and development of alternative and rapid technologies, namely RT-LAMP, ELISA, Biosensors, and GeneXpert, offer a wide range of potential options not only for SARS-CoV-2 detection but also for other viruses. This study aims to discuss the effective regional rapid detection and quantification methods in community wastewater.

Small Animal Small Animal $[^{18}F]$FDG PET Imaging for Tumor Model Study (종양 모델 연구를 위한 소동물 $[^{18}F]$FDG PET 영상화)

  • Woo, Sang-Keun;Kim, Kyeong-Min;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.1
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    • pp.1-7
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    • 2008
  • PET allows non-invasive, quantitative and repetitive imaging of biological function in living animals. Small animal PET imaging with $[^{18}F]$FDG has been successfully applied to investigation of metabolism, receptor-ligand interactions, gene expression, adoptive cell therapy and somatic gene therapy. Experimental condition of animal handling impacts on the biodistribution of $[^{18}F]$FDG in small animal study. The small animal PET and CT images were registered using the hardware fiducial markers and small animal contour point. Tumor imaging in small animal with small animal $[^{18}F]$FDG PET should be considered fasting, warming, and isoflurane anesthesia level. Registered imaging with small animal PET and CT image could be useful for the detection of tumor. Small animal experimental condition of animal handling and registration method will be of most importance for small lesion detection of metastases tumor model.

A Study on the Validation system of Detection for Biological Agents Using Real-Time PCR (실시간 중합효소 연쇄반응을 활용한 생물작용제 검증시스템 연구)

  • Cha, Younggil;Koo, Bonwoo;Kim, Seongjoo;Kim, Namil;Park, Hanoh
    • Journal of the Korea Institute of Military Science and Technology
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    • v.20 no.5
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    • pp.726-732
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    • 2017
  • Bacillus anthracis, Vibrio cholerae, Variola virus and Shigella dysenteriae are classified as category A and B biological weapons. In this study suggest that 4 genes of Bacillus anthracis, 2 genes of Vibrio cholerae, 1 gene of Variola virus and 1 gene of Shigella dysenteriae were detective 50~500 fg of target DNA per reaction using real-time PCR based assay. Also analytical specificity did not show any cross-reactivity with other related bacteria. Reliable and one reaction could be effective early diagnostic and treatment for detection of unknown samples.

Electrochemical Determination of Dopamine Based on Carbon Nanotube-Sol-Gel Titania-Nafion Composite Film Modified Electrode

  • Park, Ji-Ae;Kim, Byung-Kun;Choi, Han-Nim;Lee, Won-Yong
    • Bulletin of the Korean Chemical Society
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    • v.31 no.11
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    • pp.3123-3127
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    • 2010
  • A highly sensitive electrochemical detection method for dopamine (DA) has been developed by relying on a multiwalled carbon nanotube (CNT)-sol-gel titania-Nafion composite film modified glassy carbon (GC) electrode. The CNT-titania-Nafion/GC electrode exhibited excellent electrocatalytic activity towards DA. Therefore, the CNT-titania-Nafion/GC electrode showed improved voltammetric and amperometric responses for DA compared to those obtained with both titania-Nafion/GC and Nafion/GC electrodes. The CNT-titania-Nafion/GC electrode gave a linear response ($R^2$ = 0.999) for DA from $0.5\;{\mu}M$ to 0.5 mM with a detection limit (S/N = 3) of $0.1\;{\mu}M$ and a good sensitivity of 150 mA/M while other electrodes such as CNT-Nafion/GC, titania-Nafion/GC, and a bare GC gave a sensitivity of 89, 39, and 36 mA/M, respectively. Besides, the CNT-titania-Nafion/GC electrode displayed very fast response time within 2 s. The modified electrode showed good selectivity against ascorbic acid. The modified electrode showed good stability and reproducibility. The CNT-titania-Nafion/GC electrode was applied to the determination of DA in urine and serum samples.

Live Cell Detection of Monoclonal Antibody Light and Heavy Chain mRNAs using Molecular Beacons (분자 비컨을 이용한 살아 있는 세포에서 단일클론항체 경쇄와 중쇄 mRNA 검출에 의한 세포주 선별방법)

  • Jeong, Seunga;Rhee, Won Jong
    • KSBB Journal
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    • v.31 no.1
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    • pp.33-39
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    • 2016
  • Developing the method for the selection of animal cell line producing therapeutic monoclonal antibody (mAb) is invaluable as its market is rapidly growing. Although the quality of produced mAb is as important as quantity, however there is no method developed for the selective screening of cell lines on the basis of both quantity and quality. From recent reports, the ratio of light and heavy chain mRNAs of mAb in the cell is a key parameter for the indication of product quality. Therefore, it is obvious that developing the novel method that can detect both light and heavy chain mRNAs in single live cell will provide unprecedented opportunities in bio-industry. Here, we have constructed oligonucleotide probes, molecular beacons for the detection of light or heavy chain mRNAs, respectively, in the live cells producing mAbs. Both beacons showed increased fluorescent intensity after transient transfection of plasmid expressing mAbs analyzed by fluorometer. Flow cytometric analysis clearly demonstrated that both molecular beacons can simultaneously detect the expression of light and heavy chain mRNAs of mAb in the same cell. The technique described in the thesis provides the new direction and concept for developing the method for the smart selection of cell lines producing recombinant proteins including therapeutic mAbs.

Comparative study of linear and cyclic forms of apoptosis-targeting peptide

  • Ha, Yeong Su;Soni, Nisarg;Huynh, Phuong Tu;Lee, Byung-Heon;An, Gwang Il;Yoo, Jeongsoo
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.2 no.2
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    • pp.96-102
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    • 2016
  • Apoptosis, a genetically determined process of programmed cell death, is considered a vital component of various processes including normal cell turnover, animal development, and tissue homeostasis. It has a crucial role in many medical disorders and hence the development of non-invasive imaging tool is highly demanded. Recently, we have developed a peptide-based radioactive probe (ApoPep-1) for apoptosis detection. In that work the potential of probe for apoptosis detection was verified, however in vivo stability of radiolabeled peptide was not enough to monitor apoptosis for extended period. In current study, we prepared cyclic ApoPep-1 peptides to improve the stability of origianl linear ApoPep-1 and carried out direct comparison studies in vitro and in vivo. A targeting efficacy of newly synthesized cyclic ApoPep-1 peptide for apoptosis was confirmed in acute myocardial infarct model.