• Title/Summary/Keyword: 분자생물학적 진단

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The Detection and Diagnosis Methods of Infectious Viroids caused Plant Diseases (식물체에 감염성 질병을 유발하는 바이로이드 검출 및 진단 방법)

  • Lee, Se Hee;Kim, Yang-Hoon;Ahn, Ji-Young
    • Journal of Life Science
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    • v.26 no.5
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    • pp.620-631
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    • 2016
  • Viroids are about 250-400 base pair of short single strand RNA fragments have been associated with economically important plant diseases. Due to the lack of protein expression capacity associated with replication, it is very difficult to diagnosis viroid diseases in serological methods. For detecting viroid at plants, molecular-based techniques such as agarose gel electrophoresis, polyacrylamide gel electrophoresis (PAGE), DNA-hybridization, blotting analysis and conventional RT-PCR are reliable. Real-time RT-PCR methods that grafted on RT-PCR methods with improved confirmation methods have been also utilized. However, they are still labor-intensive, time-consuming, and require personnel with expertise. Loop-mediated Isothermal Amplification (LAMP) method is a nucleic acid amplification method under the isothermal condition. The LAMP methodology has been reported to be simple, rapid, sensitive and field applicable in detecting a variety of pathogens. The results of LAMP method can be colorized by adding a visible material such as SYBR green I, Evagreen, Calcein, Berberine and Hydroxy naphthol blue (HNB) with simple equipment or naked eyes. The combination of LAMP method and nucleic pathogens, viroids, can be used to realize simple diagnosis platform for the genetic point-of care testing system. The aim at this review is to summary viroid-caused diseases and the simple visible approach for diagnosing viroids using Loop-mediated Isothermal Amplification (LAMP) method.

Application of Methodology for Microbial Community Analysis to Gas-Phase Biofilters (폐가스 처리용 바이오필터에 미생물 군집 분석 기법의 적용)

  • Lee, Eun-Hee;Park, Hyunjung;Jo, Yun-Seong;Ryu, Hee Wook;Cho, Kyung-Suk
    • Korean Chemical Engineering Research
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    • v.48 no.2
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    • pp.147-156
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    • 2010
  • There are four key factors for gas-phase biofilters; biocatalysts(microorganisms), packing materials, design/operating techniques, and diagnosis/management techniques. Biofilter performance is significantly affected by microbial community structures as well as loading conditions. The microbial studies on biofilters are mostly performed on basis of culture-dependent methods. Recently, advanced methods have been proposed to characterize the microbial community structure in environmental samples. In this study, the physiological, biochemical and molecular methods for profiling microbial communities are reviewed, and their applicability to biofilters is discussed. Community-level physiological profile is based on the utilization capability of carbon substrate by heterotrophic community in environmental samples. Phospholipid fatty acid analysis method is based on the variability of fatty acids present in cell membranes of different microorganisms. Molecular methods using DNA directly extracted from environmental samples can be divided into "partial community DNA analysis" and "whole community DNA analysis" approaches. The former approaches consist in the analysis of PCR-amplified sequence, the genes of ribosomal operon are the most commonly used sequences. These methods include PCR fragment cloning and genetic fingerprinting such as denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis, and random amplified polymorphic DNA. The whole community DNA analysis methods are total genomic cross-DNA hybridization, thermal denaturation and reassociation of whole extracted DNA and extracted whole DNA fractionation using density gradient.

Molecular Discrimination of Dinoflagellates Cochlodinium Polykrikoides Margalef, Gyrodinium Impudicum Fraga et Bravo and Gymnodinium Catenatum Graham using RAPD-PCR Method (RAPD-PCR 방법을 이용한 Cochlodinium polykrikoides Gyrodinium impudicum, Gymnodinium catenatum의 분자생물학적 진단)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.13 no.5
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    • pp.651-657
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    • 2003
  • Randomly amplified polymorphic DNA (RAPD) analysis was used to study genetic relationships among C. polykrikoides, G. impudicum and G. catenatum, which possess similar morphological features. Four of 12 primers were selected and 59 amplification products ranged from 0.2 kb to 3.0 kb. The number of polymorphic products in C. polykrikoides, G. impudicum and G. catenatum was 16 (27.1%), 8 (13.5%), and 16 (27.1%), respectively, while 17 were monomorphic. Number of species-specific bounds was 26 (44.0%), 34 (57.6%), 26 (44.0%) in C. polykrikoides, G. impudicum and G. catenatum, respectively. The genetic similarity between C. polykrikoides and G. impudicum/G. catenatum was 0.83, whereas G. impudicum and G. catenatum was 0.78. Our results suggest that C. polykrikoides, G. impudicum and G. catenatum are extremely different on the basis of RAPD analysis, despite similarity based on their morphology. The RAPD technique appears to be efficient in detecting genetic variation in these dinoflagellates.

리포좀의 면역학적 응용

  • 정홍석;문홍모
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.11a
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    • pp.81-90
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    • 1994
  • 리포좀은 1960년대 초 Bangham등에 의해 혈액응고시 인지질의 영향에 관한 연구 중 우연히 발견하게 되었다. 즉 인지질이 물에 분산되어 2 분자막 (Bilayer)을 형성함을 알았고, 이때 생긴 폐쇄소포체를 리포좀이라 하였다. 이러한 리포좀은 원형질 막이나 세포내의 소기관의 막과 유사한 구조를 갖고 있기 때문에 이것을 이용하여 생화학자나 생물리학자들은 생체막의 모델시스템으로 이용하여 생체막의 기능 연구에 널리 이용하여 왔다. 1970년 초반부터 리포좀을 이용한 약물의 생체내 공급(Drug Delivery System, DDS)에 대하여 많은 연구가 이루어져 왔고, 지금까지도 의약품외 전달매체로 이용할려고 많은 시도를 하고 있는 실정이다. 최근에는 리포좀을 이용하여 항원을 전달하는 면역보조제(Adjuvant System)로 이용하려는 시도를 하고 있다. 현재까지 리포좀을 이용한 상품들은 화장품과 임상용 진단시 약, DNA transfection system (Lipopectin)으로 활용되고 있다.

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Access of Anti-cancer Treatment for Advanced Colon Cancer with Metastasis (전이를 동반한 진행성대장암의 항암 치료에 대한 접근)

  • Hyun Gun Kim
    • Journal of Digestive Cancer Research
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    • v.1 no.1
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    • pp.6-16
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    • 2013
  • Recent estimates for colon cancer incidence in Korea have been increased and continue to rank as the second most common in male and the third in female. Although colonoscopy has been known as the best screening tool for colon cancer, 20-25% of patients with colon cancer was diagnosed with stage IV cancer. During the past 10 years, intensive clinical studies helped to establish the value of palliative treatment for colon cancer with metastasis. The introduction of new chemotherapeutic agents such as irinotecan and oxaliplatin has led to a significant increase in tumor response and median survival. In advanced colon cancer, impressive prolongation or overall survival can be achieved through sequential application of combined systemic chemotherapy. In addition, targeted manipulation of molecular tumor mechanisms with new substances such as monoclonal antibodies against the epidermal growth factor receptor or vascular endothelial growth factor shows promising effects. Progress in the systemic treatment of colon cancer is evident, not only because of the significant increase in life expectancy in advanced colon cancer.

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Occurrence of viral nervous necrosis(VNN) in red drum(Sciaenops ocellatus) larvae (홍민어 Sciaenops ocellatus에서의 바이러스성 신경괴사증 viral nervous necrosis)

  • Kim, Jin-Do;Kim, Sok-Ryul;Jung, Sung-Ju;Kim, Young-Jin;Jung, Tae-Sung;Cho, Tae-Jin;Park, Sung-Woo;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.14 no.2
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    • pp.91-95
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    • 2001
  • Mass mortalities occurred among red drum larvae, 20 to 30 days old, culturing at hatcheries on southern costal area. No specific external signs were observed except abnormal swimming and spinal deformity. It was, however, suspected as a viral etiology due to high mass mortalities so that histopathological and molecular biological study was performed to evaluate the agent. Both vacuoles and necrosis were observed on nerve cells of brain and eye by H-E staining, and viral particles were observed on electronmicroscopic examination. On the other hand, DNA fragment, approximately 426 bps, was amplified with RT-PCR. The above results were able to diagnosis the etiological agent of mass moralities in red drum larvae as VNN(viral nervous necrosis)virus.

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Trends in Rapid Detection Methods for Food-borne pathogenic Microorganisms by Using New Technologies (신기술 이용 식중독균 신속검출법 개발 동향 분석)

  • Kim, Hyun-Joo;Kim, Yong-Soo;Chung, Myung-Sub;Oh, Deog-Hwan;Chun, Hyang-Sook;Ha, Sang-Do
    • Journal of Food Hygiene and Safety
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    • v.25 no.4
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    • pp.376-387
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    • 2010
  • Recently, speedy, convenient and easy detection technologies have been developed rapidly and on the contrary, studies on development of traditional detectors applying biochemical characteristics has gradually been decreased. This review examined trend in current studies on detection of food-borne pathogenic microorganisms in the fields of selective media, immuno-assay, Polymerase Chain Reaction (PCR), microarray, terahertz spectroscopy & imagination and so on. Most traditional methods to detect the organisms from food matrix rely on selective media and such a method have disadvantages like long time requirement and distinguishing one species only from each selective medium although they are highly economical. Various new convenient methods such as Enzyme Linked Immuno-sorbent Assay (ELISA), paper-strip kit, fluoroimmunoassay etc. have been developed. The most ideal method for detecting food-borne pathogenic microorganisms in foods should be accurate, convenient, rapid and economical. Additionally, it is needed that capabilities of quantitative analysis and automation to be applied to industries.

Immunological Studies on the Surface Antigens of Tumor Cells (종양세포 표면항원에 대한 분자면역학적 연구)

  • 김한도;김규원
    • The Korean Journal of Zoology
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    • v.32 no.2
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    • pp.142-152
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    • 1989
  • We have produced a new monoclonal antibody detecting common acute lymphoblastic leukemia antigen (CALLA) and designated as KP-22. CALIA detected by KP-22 is expressed on the all of the various cefl lines examined including common ALL. Burkitt's lymphoma, human fibroblasts and cultured normal human fibroblasts. However out of cell lines tested, a fraction of J-ALL and all of myelocytic leukemia and all other nonleukemia cell lines except for fibroblast are CALIA negative. Immunoprecipitation of solubilized 125 I-labeled membrane proteins from cultured human fibroblasts and leukemia cell lines with KP-22 revealed a major polypeptide chain with an apparent molecular weight of approximately 100 Kd and 95 Kd, respectively. Even though a microheterogeneity in terms of molecular weight between two CALLAs, the peptide mapping patterns of them &e identical indicating that such a microheterogeneity seems to be partly due to heterogeneous terminal sialic acid compositions added by a posttranslational modification process.

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Laboratory Diagnosis of Coronavirus Disease 19 (COVID-19) in Korea: Current Status, Limitation, and Challenges (국내 중증 급성 호흡기 증후군 코로나 바이러스의 검사실 내 진단: 현재, 한계점 그리고 직면한 과제)

  • Song, Gi Seon;Lee, You-Rim;Kim, Sungmin;Kim, Wontae;Choi, Jungwon;Yoo, Dahyeon;Yoo, Jungyoung;Jang, Kyung-Tae;Lee, Jaewang;Jun, Jin Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.3
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    • pp.284-295
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    • 2020
  • In December 2019, the first coronavirus disease- 2019 (COVID-19) patient was reported in Wuhan, Hubei Province, China. Since then, the number of patients who suffered severe acute respiratory syndrome caused by the novel Coronavirus (SARS-CoV-2 or 2019-nCoV) has increased dramatically in Korea. This new variant virus induces pulmonary diseases, including cough, sore throat, rhinorrhea, dyspnea, and pneumonia. Because SARS-CoV-2 is an RNA virus, real-time reverse-transcriptase PCR has been used widely to diagnose COVID-19. As the Korea Centers for Disease Prevention and Control (KCDC) and Ministry of Food & Drug Safety (MFDS) approved emergency use authorization, clinical specimens collected from COVID-19 patients and even healthy people have been clinically diagnosed by laboratory medicine. Based on a literature search, this paper reviews the epidemiology, symptoms, molecular diagnostics approved by KCDC, a current diagnosis of COVID-19 in the laboratories, the difference between molecular and serological diagnosis, and guidelines for clinical specimens. In addition, the Korean guidelines of biosafety for clinical laboratory scientists are evaluated to prevent healthcare-associated infection. The author's experience and lessons as clinical laboratory scientists will provide valuable insights to protect the domestic and international health community in this COVID-19 pandemic around the world.

Monitoring of Bifenazate Resistant Two-spotted Spider Mite, Tetranychus urticae Using Molecular Detection Method (분자학적 진단방법을 이용한 bifenazate 저항성 점박이응애 모니터링)

  • Lee, Kyu-Ri;Shin, Yun-Ho;Cho, Sun-Ran;Koo, Hyun-Na;Choi, Jang-Jeon;Ahn, Ki-Su;Kim, Gil-Hah
    • The Korean Journal of Pesticide Science
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    • v.15 no.1
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    • pp.61-67
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    • 2011
  • In 2010, two-spotted spider mite, Tetranychus urticae was collected from the rose greenhouse and apple orchards in Cheongju (CJ), Chungju (CUJ)-1, CUJ-2, Kangjin (KJ), Yesan (YS), and Yeongju (YJ). Among them, KJ and YS strain showed high resistance to bifenazate of 964.5- and 1l30-fold, respectively. The other strains showed low resistance to bifenazate. By analyzing the mitochondrial cytochrome b (cytb) sequence, G126S point mutation was detected in KJ and YS strain. Thus, G126S point mutation in the mitochondrial cytb was available molecular detection marker for selection of bifenazate resistant T. urticae. Two molecular detection methods, quantitative sequencing (QS) and PCR amplification of specific alleles (PASA) were well detected specific G126S point mutation. Therefore, these methods can be used to monitor the resistance allele in field population of T. urticae and bifenazate resistance management strategy.