• Title/Summary/Keyword: 중합효소연쇄반응법

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Monitoring of Pulmonary Tuberculosis by Polymerase Chain Reaction After Antituberculous Treatment (항결핵제 투여후 중합효소연쇄반응으로 추적한 폐결핵 환자들의 치료반응 관찰)

  • Jeon, Chang-Ho;Suh, Hun-Suk;Lee, Sang-Chae;Hyun, Dae-Sung;Ahn, Wook-Su
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.5
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    • pp.935-941
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    • 1998
  • Background: As living and dead Mycobacteria could be amplified by polymerase chain reaction(PCR), it was considered that PCR was inappropriate for the monitoring of pulmonary tuberculosis after treatment. But we found negative conversion of PCR after successful treatment. We would like to know about the negative conversion rate of PCR and its conversion time after antituberculous treatment. Methods: We collected 113 sputums from the 16 patients of pulmonary tuberculsosis visiting Catholic University Hospital of Taegu Hyosung. We consecutively tested AFB smear, AFB culture and PCR by 2 to 4 weeks after antituberculous therapy. The patients were classified according to the chest X ray findings. Results: We detected negative conversion of PCR from all 16 patients of the pulmonary tuberculosis within 30 weeks after treatment. The average negative conversion time was $16{\pm}8$ weeks. The conversion time according to the chest X -ray findings were as follows : For the 8 cases of minimum were $9{\pm}5$ weeks, 4 cases of modreate advanced were $20{\pm}8$ weeks, and 4 cases of far advanced were $23{\pm}2$ weeks. The product of PCR was gradually decreased according to the duration of treatment. Conclusions: From the results of our study, we could utilize M. tubercuosis PCR for the prediction of therapy response and monitoring of the patient with pulmonary tuberculosis after treatment.

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Diagnostic testing for Duchenne/Becker Muscular dystrophy using Dual Priming Oligonucleotide (DPO) system (Dual Priming Oligonucleotide (DPO) system을 이용한 듀시엔/베커형 근이영양증 진단법)

  • Kim, Joo-Hyun;Kim, Gu-Hwan;Lee, Jin-Joo;Lee, Dae-Hoon;Kim, Jong-Kee;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.5 no.1
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    • pp.15-20
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    • 2008
  • Purpose : Large exon deletions in the DMD gene are found in about 60% of DMD/BMD patients. Multiplex PCR has been employed to detect the deletion mutation, which frequently generates noise PCR products due to the presence of multiple primers in a single reaction as well as the stringency of PCR conditions. This often leads to a false-negative or false-positive result. To address this problematic issue, we introduced the dual primer oligonucleotide (DPO) system. DPO contains two separate priming regions joined by a polydeoxyinosine linker that results in high PCR specificity even under suboptimal PCR conditions. Methods : We tested 50 healthy male controls, 50 patients with deletion mutation as deletion-positive patient controls, and 20 patients with no deletions as deletion-negative patient controls using DPO-multiplex PCR. Both the presence and extent of deletion were verified by simplex PCR spanning the promoter region (PM) and 18 exons including exons 3, 4, 6, 8, 12, 13, 17, 19, 43-48, 50-52, and 60 in all 120 controls. Results : DPO-multiplex PCR showed 100% sensitivity and specificity for the detection a deletion. However, it showed 97.1% sensitivity and 100% specificity for determining the extent of deletions. Conclusion : The DPO-multiplex PCR method is a useful molecular test to detect large deletions of DMD for the diagnosis of patients with DMD/BMD because it is easy to perform, fast, and cost-effective and has excellent sensitivity and specificity.

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Thermal Design of PCR Chip for LOC (랩온어칩을 위한 중합효소 연쇄반응 칩의 열설계)

  • Kim, Deok-Jong;Kim, Jae-Yun;Park, Sang-Jin;Heo, Pil-U;Yun, Ui-Su
    • 연구논문집
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    • s.33
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    • pp.17-25
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    • 2003
  • In this work, thermal design of a PCR chip for LOC is systematically conducted. From the numerical simulation of a PCR chip based on the finite volume method, how to control the average temperature of a PCR chip and the temperature difference between the denaturation zone and the annealing zone is presented. The average temperature is shown to be controlled by adjusting heat input and a cooler as well as a heater is shown to be necessary to obtain three individual temperature zones for polymerase chain reaction. To reduce the time required, a heat sink for the cooler is not included in the calculation domain for the PCR chip and heat sink design is conducted separately by using a compact modeling method, the porous medium approach.

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Sex Determination used Sex Determining Region Y Gene on the Y-chromosome of Human Teeth (사람 치아 Y염색체상의 sex determining region Y(SRY)유전자를 이용한 성별감정)

  • Kim, Sei-Youn;Ahn, Jong-Mo;Ryu, Geun-Chun;Yoon, Chang-Lyuk
    • Journal of Oral Medicine and Pain
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    • v.24 no.3
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    • pp.325-333
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    • 1999
  • 최근 중합효소연쇄반응을 이용한 분자생물학적 유전자분석기술의 발달로 성염색체상의 유전좌위 증폭을 통한 성별감정이 활발히 이루어지고 있다. 그중 사람 Y염색체상에 존재하는 남성 고환의 형성을 유도하는 sex-determining region Y(SRY) gene이 규명되어 유전질환의 조기 발견이나 예방 및 태아의 성별판정 등에 응용되고 있다. 그러나, 치아는 외부 환경에 대한 저항성이 가장 높은 장기로 성별감정 등 법의치과학적 개인식별에 널리 이용되고 있음에도 불구하고, SRY 유전자를 이용하여 치아에서의 성별감정에 대한 연구는 시도된 바 없다. 따라서, 본 연구에서는 사람 치아에서 중합효소연쇄반응법을 이용한 SRY 유전자를 검출하여 성별판정에 용용하고자 하였다. 남녀 각각 20개 치아의 치수와 상아질에서 DNA를 추출하여 중합효소연쇄반응 을 시행하고 SRY 유전자를 검색한 결과, 남성에서는 치수 13개중 8개, 상아질 7개중 4개에서 SRY 유전자가 검출되었고, 여생에서는 검출되지 않았다. 이러한 결과는 중합효소연쇄반응법을 이용하여 사람 치아에서 SRY 유전자를 검색할 때, 남성판별에 유용하고 치아를 이용한 성별감정시 기존의 성별감정에 이용되고 있는 다른 유전자와 함께 SRY 유전자를 검색함으로써 성별감정의 신뢰도를 높힐 수 있을 것으로 사료된다.

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PCR-based Identification of Eubacteirum species in endodontic infection (감염 근관에서 중합효소연쇄반응법을 이용한 Eubacterium 균종의 동정)

  • Kum, Kee-Yeon;Fouad, A.F.
    • Restorative Dentistry and Endodontics
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    • v.28 no.3
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    • pp.241-248
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    • 2003
  • Asaccharolytic Eubacterium균종은 감염 근관에서의 높은 발생 빈도와 독성으로 인해 최근 많이 연구되어지고 있다. 본 연구는 24명의 환자의 감염 근관에서 얻은 22개의 PCR 산물로부터 Eggerthella lenta를 포함한 Eubacterium 균종의 빈도 및 환자의 임상 증상이나 당뇨와의 상관성을 조사한 후 얻은 자료를 토대로 다음과 같은 결과를 얻었다. 1. 22개의 표본 중에서 16개(73%)가 Eubacterium균종을 포함하고 있었으며 이 중 9개의 시편에서 Eubacterium infirmum이 검출되었다. 2. Eggerthella lenta는 어떤 시편에서도 발견되지 않았다. 3, Odds ratio analysis 결과 Eubacterium infirmum은 당뇨병과의 높은 상관성을 보여주었다(OR=9.6, P=0.04).

Detection of RSIV (Red Sea Bream Iridovirus) in the Cultured Marine Fish by the Polymerase Chain Reaction (중합효소연쇄반응 (Polymerase Chain Reaction, PCR)법을 이용한 남해안 양식 해산어의 Red Sea Bream Iridovirus (RSIV) 보유상황 확인)

  • Oh, Myung-Joo;Jung, Sung-Ju;Kim, Young-Jin
    • Journal of fish pathology
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    • v.12 no.1
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    • pp.66-69
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    • 1999
  • Occurrences of red sea bream iridovirus disease (RSIVD) in cultured marine fishes were investigated. The infection was detected by the polymerase chain reaction (PCR) used to amplify the red sea bream iridovirus (RSIV). The RSIV infection was widely distributed in fish culture farm around the south coastal area of the Korean peninsula.

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Ethidium monoazide-PCR for the detection of viable Escherichia coli in aquatic environments (수환경에서 살아 있는 대장균의 검출을 위한 ethidium monoazide-중합효소연쇄반응법)

  • Lee, Gyucheol;Kim, Hyunjeong;Lee, Byunggi;Kwon, Soonbok;Kim, Gidon;Lee, Sangtae;Lee, Chanhee
    • Journal of Korean Society of Water and Wastewater
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    • v.23 no.2
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    • pp.199-205
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    • 2009
  • It is very important to differentiate of DNA derived from live or dead bacteria within mixed microbial communities in aquatic environments. Ethidium monoazide (EMA) is a DNA intercalating agent and the treatment of EMA with strong visible light cleaves the genomic DNA of bacteria. In dead bacterial cells, EMA intercalates into the genomic DNA, induces the cleavage of DNA, and inhibits the PCR amplification. In this study, we developed the EMA-PCR and EMA real-time PCR to detect the DNA derived from viable Escherichia coli (E.coli) in mixed cultures of live and dead E.coli. The treatment of EMA, $50{\mu}g/mL$, and 650 W visible halogen light exposure for 2 minutes cleaved the genomic DNA derived from heat killed E.coli but did not those of live E.coli. EMA-PCR could detect the DNA from live E.coli in mixed culture samples of live and dead E.coli at various ratio and there was no DNA amplification in only dead E.coli cultures. Similar results were observed in EMA real-time PCR. Further studies are needed to develop various EMA-PCR methods to detect viable waterborne pathogens such as Helicobacter pylori, Giardia lamblia, and so on.