• 제목/요약/키워드: Rt

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PCR 및 RT-PCR을 이용한 하천수 중 Giardia lamblia 검출 (Detection of Giardia lamblia in River Water Samples Using PCR and RT-PCR)

  • 조은주;이목영;변승헌;한선희;안승구
    • 대한환경공학회지
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    • 제29권8호
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    • pp.904-908
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    • 2007
  • 병원성 원생동물인 지아디아 람블리아는 수인성 질병을 야기하는 주요 원인이 되고 있다. 본 연구는 PCR 및 RT-PCR 기법을 적용하여 한강본류 하천수 시료에서 사람감염성 종인 지아디아 람블리아를 동정하고, 활성 여부를 판별하여 현 표준시험방법을 보완하고자 하였다. PCR과 RT-PCR에는 지아디아의 복부 흡반 구성 유전자를 증폭하는 giardin primer를 사용하였으며, DNA/RNA 추출 및 PCR/RT-PCR 과정에서의 민감도 검사를 수행한 결과, 1포낭까지 검출가능한 것으로 나타났다. 또한 한강본류 및 유입지천 시료 48점에 적용하여 면역형광항체법과 PCR 및 RT-PCR 방법을 비교하였다. 면역형광항체법을 이용한 현미경관찰 결과 48개 시료의 지아디아 총포낭수의 평균 농도는 6.3 cysts/10 L이었고 양성율은 62.5%였으며, 속빈 포낭을 제외한 지아디아의 평균 농도는 4.5 cysts/10 L이었고 양성율은 52.1%였다. PCR 수행결과 48개 시료 중 24개(50%)의 시료에서 지아디아 람볼리아가 검출되었으며, RT-PCR 수행결과 10개(21%) 시료가 살아있는 G. lamblia를 포함한 것으로 나타났다. 본 연구를 통해 PCR/RT-PCR 기법이 지아디아 포낭을 저농도로 포함하고 있는 하천수 시료에 적용가능하며 원생동물 표준시험방법을 보완하여 종(species) 및 활성에 대한 정보를 제공할 수 있을 것으로 결과되었다.

태충(太衝)(LR3).합곡(合谷)(LI4) 전침자극(電鍼刺戟)이 뇌활성(腦活性) 변화(變化)에 미치는 영향(影響) - fMRI를 이용(利用)한 연구(硏究) - (A study on Brain activity induced by electro-acupuncture on Taechung(LR3) and Hapkok(LI4) using functional Magnetic Resonance Imaging)

  • 박태균;김영일;홍권의;임윤경;이현;이병렬
    • Korean Journal of Acupuncture
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    • 제21권2호
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    • pp.29-46
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    • 2004
  • Objectives and Methods : This study was performed to investigate the effect of electro -acupuncture at four gates(bilateral LR3 and LI4) on brain activity in normal subjects using fMRI. Results and Conclusions : 1. fMRI signal increase by electro-acupuncture at Lt. LR3 was observed in Rt. Middle frontal gyrus in group average as well as more than half of the subjects. 2. fMRI signal decreases by electro-acupuncture at Lt. LR3 were observed in Rt. Superior frontal gyrus, Rt. Middle temporal gyrus, Rt. Cingulate gyrus in group average as well as more than half of the subjects. 3. fMRI signal increases by electro-acupuncture at Lt. LI4 were observed in Lt. Superior frontal gyrus, Lt. Middle frontal gyrus, Lt. Inf. Semi-Lunar Lobule(cerebellum), Rt. Middle frontal gyrus, Rt. Cingulate gyrus in group average as well as more than half of the subjects. 4. fMRI signal decreases by electro-acupuncture at Lt. LI4 were observed in Lt. Middle frontal gyrus, Lt. Inferior frontal gyrus, Lt. Precentral gyrus and Rt. Middle frontal gyrus, Rt. Middle temporal gyrus, Rt. Precuneus, Rt. Inferior frontal gyrus, Rt. Postcentral gyrus in group average as well as more than half of the subjects. 5. fMRI signal increase by electro-acupuncture at Lt. LR3 and Lt. LI4 in group average as well as more than half of the subjects was not observed. 6. fMRI signal decreases by electro-acupuncture at Lt. LR3 and Lt. LI4 were observed in Lt. culmen(cerebellum), Lt. Cingulate gyrus와 Rt. Middle frontal gyrus, Rt. Cingulate gyrus, Rt. Inferior frontal gyrus in group average as well as more than half of the subjects. 7. fMRI signal increases by electro-acupuncture at four gates (bilateral LR3 and LI4) were observed in Lt. Middle temporal gyrus and Lt. Postcentral gyrus in group average as well as more than half of the subjects. 8. fMRI signal decrease by electro-acupuncture at four gates (bilateral LR3 and LI4) were observed in Lt. Middle frontal gyrus, Lt. Precentral gyrus, Lt. Inferior frontal gyrus, Lt. Middle temporal gyrus, Lt. Frontal sub-gyral and Rt. Tuber(cerebellum) in group average as well as more than half of the subjects.

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벼 바이러스(RSV, RBSDV)와 애멸구의 간편한 VC/RT-PCR 유전자 진단기술 (Convenient Genetic Diagnosis of Virion Captured (VC)/RT-PCR for Rice Viruses (RSV, RBSDV) and Small Brown Plant Hopper)

  • 김정수;이수헌;최홍수;조점덕;노태환;김진영
    • 식물병연구
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    • 제15권2호
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    • pp.57-62
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    • 2009
  • 우리나라 벼에 발생하는 주요 바이러스 중 애멸구에 의하여 전염하는 벼줄무의잎마름병(RSV)과 벼검은줄오갈병(RBSDV)에 대한 간편한 유전자 진단법인 VCHT-PCR 방법을 개발하였다. 벼 잎의 경우 즙액 추출 완충액은 0.5% sodium sulfite를 첨가한 0.01 M 인산완충액(pH 7.0)을 기본 완충액으로 이용하고 애멸구를 진단할 경우에는 기본 완충액에 2% PVP을 첨가하였을 때 VC/RT-PCR 진단이 잘 되었다. VC/RT-PCR을 이용한 진단에 적합한 RSV와 RBSDV 프라이머를 선발하였고 이것을 이용하여 동시진단으로 경기도 김포, 평택, 시흥지역에서 채집한 애멸구의 RSV와 RBSDV의 보독충을 쉽고 경제적으로 진단 할 수 있었다. ELISA에 의한 진단결과와 비교할 때 세 지역을 합하여 RSV에 대한 평균 보독충률은 9.2%로 동일하였으나 보독충 중 일부가 ELISA와 VC/RT-PCR 두 방법에 의한 진단결과가 다르게 나온 것은 RSV의 혈청학적, 유전적 계통 존재 가능성을 제시하고 있다. 포장에서 채집한 애멸구의 VC/RT-PCR 진단효율은 RSV와 RBSDV를 동시에 진단하므로 써 RSV만 진단이 가능한 ELISA 결과 보다 3.2% 높았다.

Development of a Rapid Detection Method for Potato virus X by Reverse Transcription Loop-Mediated Isothermal Amplification

  • Jeong, Joojin;Cho, Sang-Yun;Lee, Wang-Hyu;Lee, Kui-jae;Ju, Ho-Jong
    • The Plant Pathology Journal
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    • 제31권3호
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    • pp.219-225
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    • 2015
  • The primary step for efficient control of viral diseases is the development of simple, rapid, and sensitive virus detection. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has been used to detect viral RNA molecules because of its simplicity and high sensitivity for a number of viruses. RT-LAMP for the detection of Potato virus X (PVX) was developed and compared with conventional reverse transcription polymerase chain reaction (RT-PCR) to demonstrate its advantages over RT-PCR. RT-LAMP reactions were conducted with or without a set of loop primers since one out of six primers showed PVX specificity. Based on real-time monitoring, RT-LAMP detected PVX around 30 min, compared to 120 min for RT-PCR. By adding a fluorescent reagent during the reaction, the extra step of visualization by gel electrophoresis was not necessary. RT-LAMP was conducted using simple inexpensive instruments and a regular incubator to evaluate whether RNA could be amplified at a constant temperature instead of using an expensive thermal cycler. This study shows the potential of RT-LAMP for the diagnosis of viral diseases and PVX epidemiology because of its simplicity and rapidness compared to RT-PCR.

Comparison of Molecular Assays for the Rapid Detection and Simultaneous Subtype Differentiation of the Pandemic Influenza A (H1N1) 2009 Virus

  • Lee, Mi Kyung;Kim, Hye Ryoun
    • Journal of Microbiology and Biotechnology
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    • 제22권8호
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    • pp.1165-1169
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    • 2012
  • In April 2009, the H1N1 pandemic influenza virus emerged as a novel influenza virus. The aim of this study was to compare the performances of several molecular assays, including conventional reverse transcription polymerase chain reaction (RT-PCR), two real-time reverse transcription (rRT)-PCRs, and two multiplex RTPCRs. A total of 381 clinical specimens were collected from patients (223 men and 158 women), and both the Seeplex RV7 assay and rRT-PCR were ordered on different specimens within one week after collection. The concordance rate for the two methods was 87% (332/381), and the discrepancy rate was 13% (49/381). The positive rates for the molecular assays studied included 93.1% for the multiplex Seeplex RV7 assay, 93.1% for conventional reverse transcription (cRT)-PCR, 89.7% for the multiplex Seeplex Flu ACE Subtyping assay, 82.8% for protocol B rRT-PCR, and 58.6% for protocol A rRT-PCR. Our results showed that the multiplex Seeplex assays and the cRT-PCR yielded higher detection rates than rRT-PCRs for detecting the influenza A (H1N1) virus. Although the multiplex Seeplex assays had the advantage of simultaneous detection of several viruses, they were time-consuming and troublesome. Our results show that, although rRT-PCR had the advantage, the detection rates of the molecular assays varied depending upon the source of the influenza A (H1N1)v virus. Our findings also suggest that rRT-PCR sometimes detected virus in extremely low abundance and thus required validation of analytical performance and clinical correlation.

Retroperitoneal liposarcoma: the role of adjuvant radiation therapy and the prognostic factors

  • Lee, Hong Seok;Yu, Jeong Il;Lim, Do Hoon;Kim, Sung Joo
    • Radiation Oncology Journal
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    • 제34권3호
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    • pp.216-222
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    • 2016
  • Purpose: To evaluate the benefit of adjuvant radiation therapy (RT) for retroperitoneal liposarcoma (RPLS) following gross tumor removal. Materials and Methods: We reviewed 77 patients with primary RPLS surgically treated between January 2000 and December 2013. Cases with gross residual disease were excluded. Tumor grade was evaluated according to the French Federation of Cancer Centers Sarcoma Group (FNCLCC) system. Adjuvant RT was delivered to 32 patients (42%) using external beam RT alone. Median follow-up time was 36 months (range, 5 to 169). Results: Among 77 patients, 33 (43%) presented with well-differentiated, 31 (40%) with de-differentiated, 8 (10%) with myxoid/round and 4 (5%) with pleomorphic morphology. The RT group included less well-differentiated subtype than surgery group (28% vs. 53%). During follow up, 34 patients (44%) showed local recurrence. Local recurrence rate was lower in the RT group (38%) compared to the surgery group (49%). The 3-year local control rate (LC) was 55.6%, and the 3-year overall survival (OS) was 82.1%. Tumor histology and FNCLCC grade were significantly associated with local recurrence. There was no statistical significance of adding adjuvant RT in LC (p = 0.312). However, patients with tumor histology other than well-differentiated subtype showed marginally decreased local recurrence rate after adjuvant RT (3-year LC, RT 43.9% vs. no RT 35.3%; p = 0.087). Conclusion: RPLS patients receiving RT experienced less local recurrence. We suggest that the addition of adjuvant RT may be related to improvement of LCs, especially in patients with non-favorable histologic subtypes.

Role of radiotherapy for pancreatobiliary neuroendocrine tumors

  • Lee, Jeongshim;Choi, Jinhyun;Choi, Chihwan;Seong, Jinsil
    • Radiation Oncology Journal
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    • 제31권3호
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    • pp.125-130
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    • 2013
  • Purpose: We investigated the role of radiotherapy (RT) for pancreatobiliary neuroendocrine tumors (PB-NETs). Materials and Methods: We identified 9 patients with PB-NETs who received RT between January 2005 and March 2012. Of these 9 patients, 4 were diagnosed with NETs in the pancreas and 5 were diagnosed with NETs in the gallbladder. All patients received RT to the primary tumor or resection bed with a median total irradiation dose of 50.4 Gy, with or without chemotherapy. Results: The tumor response rate and tumor control rate in the RT field were 60% and 100 %, respectively. All 4 patients who underwent surgery had no evidence of disease in the RT field. Of the 5 patients who received RT to the primary gross tumor, 1 had complete response, 2 had partial response, and 2 had stable disease in the RT field. The median time to progression was 11 months. Of the 9 patients, four patients had no progression, and 5 patients had progression of disease (locoregional, 2; distant, 2; locoregional/distant, 1). Of the 4 patients without progression, 3 were treated with RT in adjuvant or neoadjuvant setting, and one received RT to primary tumor. One patient experienced radiation-induced duodenitis at 3 months after concurrent chemoradiation without treatment-related mortality. Conclusion: RT can yield local control for advanced PB-NETs. RT should be considered an essential part of multimodality treatment in management of advanced PB-NETs.

폐암 환자에서 방사선치료가 세포성 면역반응에 미치는 영향 (The Effect of Radiation Therapy on Cellular Immune Response in Patients with Squamous Cell Lung Carcinoma)

  • 어수택;김철현;정연태;김용훈;박춘식;이희발;허승재
    • Tuberculosis and Respiratory Diseases
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    • 제38권1호
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    • pp.25-33
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    • 1991
  • The immune staus is known to be decreased in malignant disease and radiation therapy (RT), used as a therapeutic tool, further decrease this-attenuated immune status. We measured the number of peripheral lymphocytes, its subsets and lymphoblast transformation for PPD, PHA, monoclonal antibodies including anti-CD3 and anti-CD2 before and after RT in 19 patients with squamous cell lung cancer to search the fine mechanism behind the RT-induced attenuation of lymphoblast transformtion for mitogens and antigen. The results were as follows; 1) The number of lymphocytes and its subsets decreased significantly after RT, but the percentages of lymhocyte subsets did not change aftr RT except interleukin-2 receptor positive T lymphocytes. 2) The function of lymphoctes, measured by lymphoblast tranformation for PHA and PPD, decrased after RT and the compositions of PBMC used for lymphoblast transformtion were not different before and after RT. 3) The mitosis of lymphocytes to anti-CD2 or anti-CD3 decreased significantly after RT. And IL-2 plus anti-CD3 increased the mitosis than that of anti-CD3 only after RT, but before RT there was no difference. In conclusion, we suggested the fine mechanism behind the RT-induced attenuation of immune response might be the dysfunction of lymphocytes in terms of impaired synthesis of IL-2 rather than the decrease of circulating lymphocyte numbers.

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간이 조즙액 추출법을 이용한 RT-PCR 방법에 의한 오이 모자이크 바이러스의 검정 (Detection of Cucumber Mosaic Virus by RT-PCR Using a Simple and Rapid Crude Sap Extraction Method)

  • 이상용;홍진성;이진상;최장경
    • 한국식물병리학회지
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    • 제12권4호
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    • pp.432-436
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    • 1996
  • 역전사 중합효소 연쇄반응(RT-PCR)을 이용하여 담배(Nicotiana glutinosa)에 증식시킨 7종의 오이 모자이크 바이러스(CMV)를 검정하였다. RT-PCR을 위한 간단하고 신속한 바이러스 핵산의 조즙액 추출법이 개발되었으며, CMV 외피단백질 유전자 부위를 기초로 하여 제작한 20개의 염기로 구성된 primer를 사용하여 RT-PCR을 실시한 결과, 약 490 염기쌍의 DNA 단편들이 이병식물의 조즙액으로부터 증폭되었다. EcoRI 및 MspI을 이용한 RT-PCR 산물의 분석에 의하여, 공시한 7종의 바이러스는 모두 CMV subgroup I으로 동정되었다. Ouchterlony 한천젤 이중 확산법을 이용한 항혈청 검정에서도 7종의 바이러스 모두 CMV-Y의 항혈청과 단일의 침강선을 형성하였다.

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Multiplex RT-PCR 기법을 이용한 소의 로타바이러스, 코로나바이러스 및 설사병바이러스의 동시진단 (A Study on Simultanious Detection of Bovine Rotavirus, Coronavirus and Virai Diarrhea virus by Multiplex RT-PCR)

  • 노환국;이장형
    • 현장농수산연구지
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    • 제5권1호
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    • pp.57-63
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    • 2003
  • The bovine rotavirus(BRV), bovine coronavirus(BCV) and bovine viral diarrhea virus(BVDV) are main viruses of bovine viral diarrhea disease. These viruses could be rapidly amplified by the reverse transcriptase polymerase chain reaction(RT-PCR). This study was conducted to develop rapid and accurate diagnostic methods of these viral diseases by multiplex RT-PCR. Specific primers were designed based on the sequences reported by Chang KO et. al. (1997) and Schroeder BA, et. al. (1990), RNA were prepared from the cultured viruses, first-stranded DNAs were synthesised by reverse transcriptase. PCR were conducted to amplify specific regions of the viruses by multiplex. Three bands such as 1,062bp for BRV, 458bp for BCV, and 300bp for BVDV were successfully produced by multiplex RT-PCR. In conclusion, this result suggested that these viruses could be diagnosed rapidly and accurately by multiplex RT-PCR.