• 제목/요약/키워드: fluorescence diagnosis

검색결과 138건 처리시간 0.025초

Fluorometric Detection of Low-Abundance EGFR Exon 19 Deletion Mutation Using Tandem Gene Amplification

  • Kim, Dong-Min;Zhang, Shichen;Kim, Minhee;Kim, Dong-Eun
    • Journal of Microbiology and Biotechnology
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    • 제30권5호
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    • pp.662-667
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    • 2020
  • Epidermal growth factor receptor (EGFR) mutations are not only genetic markers for diagnosis but also biomarkers of clinical-response against tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). Among the EGFR mutations, the in-frame deletion mutation in EGFR exon 19 kinase domain (EGFR exon 19-del) is the most frequent mutation, accounting for about 45% of EGFR mutations in NSCLCs. Development of sensitive method for detecting the EGFR mutation is highly required to make a better screening for drug-response in the treatment of NSCLC patients. Here, we developed a fluorometric tandem gene amplification assay for sensitive detection of low-abundance EGFR exon 19-del mutant genomic DNA. The method consists of pre-amplification with PCR, thermal cycling of ligation by Taq ligase, and subsequent rolling circle amplification (RCA). PCR-amplified DNA from genomic DNA samples was used as splint DNA to conjugate both ends of linear padlock DNA, generating circular padlock DNA template for RCA. Long stretches of ssDNA harboring multiple copies of G-quadruplex structure was generated in RCA and detected by thioflavin T (ThT) fluorescence, which is specifically intercalated into the G-quadruplex, emitting strong fluorescence. Sensitivity of tandem gene amplification assay for detection of the EGFR exon 19-del from gDNA was as low as 3.6 pg, and mutant gDNA present in the pooled normal plasma was readily detected as low as 1% fraction. Hence, fluorometric detection of low-abundance EGFR exon 19 deletion mutation using tandem gene amplification may be applicable to clinical diagnosis of NSCLC patients with appropriate TKI treatment.

Detecting Incipient Caries Using Front-illuminated Infrared Light Scattering Imaging

  • Kim, Ji-Young;Ro, Jung-Hoon;Jeon, Gye-Rok;Kim, Jin-Bom;Ye, Soo-Young
    • Transactions on Electrical and Electronic Materials
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    • 제13권6호
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    • pp.310-316
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    • 2012
  • A new method for early caries diagnosis was proposed and tested through a home-made optical examination system that used quantitative light fluorescence (QLF) and digital imaging fiber optic transillumination (FOTI) (DIFOTI), with light sources across a wide spectral range, from 350 nm to 1,000 nm. The front-illuminated infrared light scattering image (FIR) showed similar diagnostic abilities to that of DIFOTI. The FIR method was invented based on the observation that caries lesions lose the high transmittance and low scattering properties of benign enamel tissue. There are various methods for the early diagnosis of caries, such as visual examination, exploration, X-ray radiography, QLF, FOTI, and infrared fluorescence (diagnodent). Among them, methods based on optical properties are regarded as having the most potential. A comparative study was performed between the FOTI, QLF, diagnodent, optical coherence tomography, and FIR scattering image methods, using 20 extracted teeth samples with early caries. A scale of lesion measurement based on optical image contrast was proposed. The statistical analysis showed a significant correlation between the DIFOTI and FIR methods (r = 0.35, p < 0.05). However, the QLF and diagnodent methods showed little association with FIR images, as they have different detection principles as compared with FIR. Tomographic images obtained by OCT, using 1,330 nm super luminescent LED as a gold standard of tooth structure, verified that the FOTI and FIR results correctly represented the lack of homogeneity in dental tissue. The newly proposed FIR method attained similar diagnostic results to those of FOTI, but with an easier approach.

Label/Quencher-Free Detection of Exon Deletion Mutation in Epidermal Growth Factor Receptor Gene Using G-Quadruplex-Inducing DNA Probe

  • Kim, Hyo Ryoung;Lee, Il Joon;Kim, Dong-Eun
    • Journal of Microbiology and Biotechnology
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    • 제27권1호
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    • pp.72-76
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    • 2017
  • Detection of exon 19 deletion mutation in the epidermal growth factor receptor (EGFR) gene, which results in increased and sustained phosphorylation of EGFR, is important for diagnosis and treatment guidelines in non-small-cell lung cancer. Here, we have developed a simple and convenient detection system using the interaction between G-quadruplex and fluorophore thioflavin T (ThT) for discriminating EGFR exon 19 deletion mutant DNA from wild type without a label and quencher. In the presence of exon 19 deletion mutant DNA, the probe DNAs annealed to the target sequences were transformed into G-quadruplex structure. Subsequent intercalation of ThT into the G-quadruplex resulted in a light-up fluorescence signal, which reflects the amount of mutant DNA. Due to stark differences in fluorescence intensity between mutant and wild-type DNA, we suggest that the induced G-quadruplex structure in the probe DNA can report the presence of cancer-causing deletion mutant DNAs with high sensitivity.

ICP 광원 시스템의 Ne:Xe, Ne:Ar 혼합가스의 단일탐침법을 이용한 플라즈마 진단 (Plasma Diagnosis of Ne:Xe, Ne:Ar Mixed Gases by Single Langmuir Probe in Inductively Coupled Plasma Light Source System)

  • 최용성;이우기;;이경섭;이상헌
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2006년도 영호남 합동 학술대회 및 춘계학술대회 논문집 센서 박막 기술교육
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    • pp.91-95
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    • 2006
  • In whole world consciousness of environment maintenance have increased very quickly for the end of the 20th century. To use and disuse toxic substances have been controled at the field of industry. Also the field of lighting source belong to environmental control. And in the future the control will be strong. In radiational mechanism of fluorescence lamp mechanism is the worst environmental problem. In radiational mechanism of fluorescence lamp mercury is the worst environmental problem root. In the mercury free lighting source system the Xe gas lamp is one type. And the Ne:Xe mixing gas lamp improvements firing voltage of Xe gas lamp. Purpose and subject of this study are understand, efficiency, ideal of Ne:Xe plasma which mercury free lamp. Before ICP was designed, basic parameters of plasma, which are electron temperature and electron density, were measured and calculated by Langmuir probe data. Property of electron temperature and electron density were confirmed by changing ratio of Ne:Xe.

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치아우식증의 조기진단을 위한 고감도 우식활성검사 모형개발 (DEVELOPMENT OF HIGH SENSITIVE MODEL OF CARIES ACTIVITY TEST FOR EARLY DIAGNOSIS OF DENTAL CARIES)

  • 이상호;이창섭
    • 대한소아치과학회지
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    • 제27권1호
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    • pp.169-179
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    • 2000
  • 초기 우식병소와 건전한 치질은 광학적 특성 이 다르다는 사실에 근거하여 치과 치료용으로 개발된 가시광선 영역의 아르곤 레이저의 형광현상, 즉 물체에 조사되어 반사된 및은 물체의 광학적 특성에 따라 파장이 달라지는 특성을 이용하여, 우식 치질과 건전한 치질간의 광학적 특성의 차이를 아르곤 레이저 빛을 통한 색조의 차이로 유도 전환하는 시스템을 개발하고 이를 이용하여 초기 치아우식 병소를 조기에 시각적으로 탐지하여 data화 하는 새로운 우식활성검사법을 개발하는데 본 연구의 목적이 있다. 연구는 3단계로 나누어 1단계는 아르곤 레이저 형광법의 초기 우식병소 탐지능력을 평가하였으며 2단계는 임상에서 아동들을 대상으로 아르곤 레이저 형광법을 이용하여 초기 우식병소가 있는 치아의 갯수를 집계하고 이 수치에 대한 우식활성검사법인 $Cariescreen^{(R)}$ test, 그리고 일종의 구강환경검사인 우식경험치아의 수(dDfFtT) 검사와의 상관성을 평가하였다. 3단계는 이와 같은 자료를 기준으로 하여 레이저 형광법을 이용하여 검사할 수 있는 새로운 우식활성검사 모델을 만들고 이와 같은 검사법의 민감도, 타당도, 진단력 등을 분석하여 진단학적 측면에서 활용 가능성을 평가하고자 하였으며 그 결과는 다음과 같다. 초기 우식병소의 조직학적 깊이가 증가될수록 레이저 형광법을 이용하여 측정한 광밀도가 증가하였으며 이들 사이에는 높은 상관관계 (=0.7015)가 있었다. 또한 통계학적으로 회귀분석을 시행한 결과 병소의 깊이와 광밀도와 사이에는 정비례 관계의 직선방정식 관계를 나타내었다. 레이저 형광법을 이용한 우식활성검사법은 $Cariescreen^{(R)}$ test 그리고 dDfFtT 검사 등과 높은 상관관계를 나타냈다. 또한 레이저 형광법을 이용한 우식활성 검사법은 dDfFtT검사를 기준으로 48%의 민감도, 52%의 특이도, 45%의 진단력을 보였으며 $Cariescreen^{(R)}$ test를 기준으로 48%의 민감도, 51%의 특이도, 36%의 진단력을 보였다. 이상의 결과를 종합하면 레이저 형광법을 이용하여 측정한 초기 우식병소의 광밀도는 실제 초기우식병소의 조직학적병소의 깊이를 평가하는데 높은 진단학적 신뢰성을 보여주고 있으며 레이저 형광법을 이용한 우식활성검사는 진단학적 지표를 기준으로 볼 때, 그리고 검사절차와 시간, 비용 등의 면으로 볼 때 향후 임상적으로 활용 가능성이 높은 것으로 사료된다.

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분자영상의 적용분야 및 전망 (Application and Prospects of Molecular Imaging)

  • 최규락;이상복
    • 한국방사선학회논문지
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    • 제8권3호
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    • pp.123-136
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    • 2014
  • 본 논문에서는 분자영상을 분류하고 적용 분야와 미래를 예측해 보고자 하였다. 분자영상은 생체 내에서 분자수준과 세포수준에서 일어나는 변화를 영상화하는 것으로써 분자세포생물학과 첨단영상기술이 발전하여 접목된 새로운 분야이다. 분자영상은 형광, 생물발광, SPECT, PET, MRI, Ultrasound 등의 영상 기법들을 이용하여 유전자 치료 모니터링, 세포추적, 세포 치료 모니터링, 항체영상, 약제 개발, 분자 상호작용 영상, 근적외선 형광 물질을 이용한 암 형광 영상, Bacteria 를 이용한 종양 표적 영상, 치료효과 조기 평가, 치료 효과 예측 등에 적용되고 있다. 분자 영상의 미래는 분자세포 생물학, 유전학, 화학, 약학, 물리학, 전산학, 의공학, 핵의학, 영상의학, 임상의학 등 여러 학문 분야가 융합되어 상호협조와 공동연구를 통하여 발전해 나갈 것이다. 분자영상의 태동으로 미래의 의료의 모습은 질병의 조기진단과 개인 맞춤형 치료가 가능하게 될 것이다.

LIF를 이용한 Hg-Ar(1Torr)의 플라즈마 밀도 측정 (Plasma Density Measurement of the Hg-Ar(1Torr) by LIF Method)

  • 이종찬;박대희;양종경
    • 대한전기학회논문지:전기물성ㆍ응용부문C
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    • 제54권5호
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    • pp.213-217
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    • 2005
  • In this paper, we introduced a LIF measurement method and summarized the theoretical side. When an altered wavelength of laser and electric power, lamp applied electric power, we measured the relative density of the metastable state in mercury after observing a laser induced fluorescence signal of 404.8nm and 546.2nm, and confirmed the horizontal distribution of plasma density in the discharge lamp. Due to this generation, the extinction of atoms in a metastable state occurred through collision, ionization, and excitation between plasma particles. The density and distribution of the metastable state depended on the energy and density of plasma particles, intensely This highlights the importance of measuring density distribution in plasma electric discharge mechanism study The results confirmed the resonance phenomenon regarding the energy level of atoms along a wavelength change, and also confirmed that the largest fluorescent signal in 436nm, and that the density of atoms in 546.2nm ($6^3S_1 {\to} 6^3P_2$ ) were larger than 404.8nm ($6^3S_1 {\to} 6^3P_1$). According to the increase of lamp applied electric power, plasma density increased, too. When increased with laser electric power, the LIF signal reached a saturation state in more than 2.6mJ. When partial plasma density distribution along a horizontal axis was measured using the laser induced fluorescence method, the density decreased by recombination away from the center.

1p36 deletion syndrome confirmed by fluorescence in situ hybridization and array-comparative genomic hybridization analysis

  • Kang, Dong Soo;Shin, Eunsim;Yu, Jeesuk
    • Clinical and Experimental Pediatrics
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    • 제59권sup1호
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    • pp.14-18
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    • 2016
  • Pediatric epilepsy can be caused by various conditions, including specific syndromes. 1p36 deletion syndrome is reported in 1 in 5,000-10,000 newborns, and its characteristic clinical features include developmental delay, mental retardation, hypotonia, congenital heart defects, seizure, and facial dysmorphism. However, detection of the terminal deletion in chromosome 1p by conventional G-banded karyotyping is difficult. Here we present a case of epilepsy with profound developmental delay and characteristic phenotypes. A 7-year-and 6-month-old boy experienced afebrile generalized seizure at the age of 5 years and 3 months. He had recurrent febrile seizures since 12 months of age and showed severe global developmental delay, remarkable hypotonia, short stature, and dysmorphic features such as microcephaly; small, low-set ears; dark, straight eyebrows; deep-set eyes; flat nasal bridge; midface hypoplasia; and a small, pointed chin. Previous diagnostic work-up, including conventional chromosomal analysis, revealed no definite causes. However, array-comparative genomic hybridization analysis revealed 1p36 deletion syndrome with a 9.15-Mb copy loss of the 1p36.33-1p36.22 region, and fluorescence in situ hybridization analysis (FISH) confirmed this diagnosis. This case highlights the need to consider detailed chromosomal study for patients with delayed development and epilepsy. Furthermore, 1p36 deletion syndrome should be considered for patients presenting seizure and moderate-to-severe developmental delay, particularly if the patient exhibits dysmorphic features, short stature, and hypotonia.

LIF를 이용한 Hg-Ar의 플라즈마 밀도 측정 (Plasma Density Measurement of Hg-Ar by LIF Method)

  • 최용성;황종선;박계춘;송민종;김형곤;이경섭
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2006년도 추계학술대회 논문집 광주전남지부
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    • pp.27-32
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    • 2006
  • In this paper, we introduced a LIF measurement method and summarized the theoretical side. When an altered wavelength of laser and electric power, lamp applied electric power, we measured the relative density of the metastable state in mercury after observing a laser induced fluorescence signal of 404.8nm and 546.2nm, and confirmed the horizontal distribution of plasma density in the discharge lamp. Due to this generation, the extinction of atoms in a metastable state occurred through collision, ionization, and excitation between plasma particles. The density and distribution of the metastable state depended on the energy and density of plasma particles, intensely. This highlights the importance of measuring density distribution in plasma electric discharge mechanism study. The results confirmed the resonance phenomenon regarding the energy level of atoms along a wavelength change, and also confirmed that the largest fluorcscent signal in 436nm, and that the density of atoms in 546.2nm ($6^3S_1{\rightarrow}6^3P_2$) were larger than 404.8nm ($6^3S_1{\rightarrow}6^3P_2$). According to the increase of lamp applied electric power, plasma density increased, too. When increased with laser electric power, the LIF signal reached a saturation state in more than 2.6mJ. When partial plasma density distribution along a horizontal axis was measured using the laser induced fluorescence method, the density decreased by recombination away from the center.

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서버 클라이언트 기반의 실시간 마이크로칩 형광 이미지 분석 시스템 개발 (Development of a real-time Analysis System of Microchip Fluorescence Images based on Server-Client)

  • 조미경;심재술
    • 한국정보통신학회논문지
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    • 제17권5호
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    • pp.1239-1244
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    • 2013
  • 임상 의료 분야에서 질병 진단 및 치료를 위해서는 분자 수준(프로틴, DNA 등)의 크기 뿐만 아니라, 세포 수준에 대한 분석이 필요하다. 많은 경우 실험 샘플이 시간에 따라 변질되기 때문에 정확한 분석을 위해서는 빠른 분석과 실시간 데이터가 필요하다. 본 연구에서는 나노 마이크로 크기의 세포내 단백질이나 DNA의 변화 과정 등을 촬영할 수 있는 3차원 형광 관측 장치를 제작하고 이로부터 얻은 형광 이미지를 실시간 통합 관리 및 분석하기 위한 서버 클라이언트 기반의 형광 이미지 분석 시스템을 구축하였다. 시스템은 형광 관측 장치와 소프트웨어 그리고 형광 이미지를 실시간으로 분석할 수 있는 모바일 프로그램으로 구성된다. 개발된 시스템은 의료인이 시공간의 제약 없이 응급환자의 샘플에서 획득한 형광이미지를 실시간으로 전송받아 분석 및 진단을 내릴 수 있도록 해 주므로 유비쿼터스 헬스 구현에 활용할 수 있다.