• Title/Summary/Keyword: 연쇄반응

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Detection of Theileria sergenti in Bovine Lymphocyte by Polymerase Chain Reaction (PCR 기법을 이용한 소 Lymphocyte 내 Theileria sergenti의 검출)

  • 박진호;이승옥;권오덕;이주묵
    • Journal of Veterinary Clinics
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    • v.15 no.1
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    • pp.146-150
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    • 1998
  • 소 림프구내의 Theileria sergenti감염을 확인하기 위하여 T. sergenti감염혈액에서 림프구를 분리한 후 중합효소연쇄반웅을 실시하였다. 또한,분리한 림프구내의 T. sergenti감 염을 증명하기 위하여 IFA test와 acridine orange stain을 실시하였다. 그 결과 다음과 같은 성적을 얻을 수 있었다. T. sergenti 감염헐액의 전혈과 림프구를 각각 생리식염수로 2배율 연 속회석하여 중합효소연쇄반응을 실시한 결과, 림프구내에서는 1,024배 회석배율까지 T. sergenti의 genomic DNA가 중폭되었으며, 전혈내에서는 256배 회석배율까지 증폭되었다. 그리 고 중합효소연쇄반응으로 T. sergenti 감염이 확인된 림프구를 이용하여 IFA test와 acridine orange 염색을 실시한 결과, 림프구내에 T. sergenti가 존재하는 것을 증명할 수있었다. 한편, 중합효소연쇄반응을 이용한 림프구내의 T. sergenti 감염의 진단 유용성을 확인하기 위하여 전 북지역에서 사육중인 소 16두를 대상으로 이들의 혈액으로 PCR 증폭을 실시하였다. 그 결과 전혈에서 genomic DNA를 취한 경우에는 3두(18.8%)만이, 그리고 림프구에서 genomic DNA를 취한 경우에는 11두(68.8%)의 소에서 T. sergenti DNA의 증폭을 관찰할 수 있었다.

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Improving Interactivity via Chained Priority Boosting for Android Smartphone (연쇄적 우선순위 상승 기법에 의한 안드로이드 스마트폰의 사용자 반응성 향상)

  • Lee, Joonghyun;Huh, Sungju;Hong, Seongsoo
    • Proceedings of the Korean Society of Computer Information Conference
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    • 2013.01a
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    • pp.1-2
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    • 2013
  • 본 논문에서는 안드로이드의 고질적인 문제점인 사용자 반응성 문제 해결을 위한 연구를 소개한다. 특히 여러 응용들이 동시에 수행되는 경우 대화형 응용이 다른 응용들에 밀려 원하는 만큼 CPU를 얻지 못하는 상황에서 발생하는 반응지연 문제에 초점을 맞추고 이를 극복하기 위한 연쇄적 우선순위 상승 기법을 제시한다. 이 기법은 대화형 웅용뿐만 아니라 기존 연구에서 고려하지 않은 터치 관련 이벤트 처리 스레드들과 대화형 응용의 자식 스레드들의 우선순위를 연쇄적으로 상향시킴으로써 터치에 대한 응답시간을 줄인다. 본 논문에서는 제안한 기법을 상용 스마트폰에 적용하여 유용성을 검증하였다. 실험 결과에 따르면 기존 안드로이드에 제안한 기법을 적용한 경우 평균반응시간이 기존의 31.91%로 감소하였다.

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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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Comparative Study of Target Genes and Protocols by Country for Detection of SARS-CoV-2 based on Polymerase Chain Reaction (PCR) (중합효소 연쇄반응 기반의 코로나-19 바이러스 검출법에 대한 국가별 목표 유전자 및 프로토콜 비교 연구)

  • Kim, Jin-Hee
    • The Journal of the Korea Contents Association
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    • v.21 no.1
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    • pp.465-474
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    • 2021
  • Corona-19, a disease caused by 'Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)', was declared a global pandemic by the World Health Organization (WHO) in March 2020, and a real-time polymerase chain reaction test is performed as a diagnostic test for screening and confirmation in most countries. However, not only the target genes and protocols differ by countries, but also the procedures for reading the diagnosis results are diverse, so the criteria for confirmed patients differ by country. Therefore, in this review, we discussed the target genes, test techniques, and diagnostic criteria for each country notified by WHO. And the specificity and sensitivity, limits of detection, positive and negative controls, false positive bacteria candidates, and specimens, and the specifics of the control setting were also described. In addition, the characteristics of Korea's test were compared to each country's one. Finally, in order to obtain the same diagnosis result for SARS-CoV-2 in the future, standardized diagnosis methods and result interpretations for Corona-19 diagnosis were proposed.

Development of Real-time PCR Assays for Detection of Dirofilaria immitis from Infected Dog Blood (심장사상충에 감염된 개의 혈액에서 심장사상충 유전자를 검출할 수 있는 실시간 중합효소연쇄반응 기법 개발)

  • Oh, In Young;Kim, Kyung Tae;Jun, Jin Hyun;Shin, Jae-Ho;Sung, Ho Joong
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.88-93
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    • 2016
  • Dirofilaria immitis is a filarial nematode parasite that causes cardiopulmonary dirofilariasis in dogs. The purpose of this study was the development of real-time PCR assays for efficient detection of D. immitis. The D. immitis-specific primers confirmed in our previous study and a newly designed TaqMan probe were used for quantitative diagnostics. First, SYBR Green real-time PCR was performed using the specific primers and serially diluted genomic DNA or plasmid DNA, and melting curve analyses were performed after amplification. The melting curve showed one specific peak in each of the genomic and plasmid DNA reactions, suggesting that the primers specifically amplify the D. immitis cytochrome c oxidase subunit I gene. Comparison of SYBR Green and TaqMan real-time PCR using serially diluted plasmid DNA showed higher efficiency and specificity with TaqMan real-time PCR. The real-time PCR assays developed in this study will provide improved diagnostic methods to overcome the limitations of conventional diagnostic tools and facilitate more rapid and accurate diagnoses.

Two-enzyme coupled fluorometric assay of urinary dipeptidase (이원효소 연쇄반응의 형광분석에 의한 Urinary Dipeptidase의 활성도 측정)

  • Park, Haeng Soon;We, Jeoung Soon
    • Analytical Science and Technology
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    • v.8 no.3
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    • pp.359-364
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    • 1995
  • Urinary dipeptidase(Udpase) was assayed by fluorometric analysis of NADH which was produced from an indicator enzyme, L-alanine dehydrogenase. The reaction mixture was consisted of a dipeptide(L-ala-L-ala), ${\beta}-NAD^+$, L-alanine dehydrogenase in 12.5 mM sodium carbonate buffer, pH 9.0, and urinary dipeptidase which initiated the reaction. The fluorescence intensity of NADH was measured as a function of time with the excitation wavelength at 340nm and emission at 460nm. Comparison of this fluorometric method with the conventional spectrophotometric method utilizing glycyldehydrophenylalanine(Gdp) as substrate provided the correlation coefficient of 0.996 and increased the sensitivity more than ten times.

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Detection of Enterohemorrhagic Escherichia coli O157:H7 Strains Using Multiplex Polymerase Chain Reaction (Multiplex PCR을 이용한 장출혈성 대장균 O157:H7의 검출)

  • 엄용빈;김종배
    • Biomedical Science Letters
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    • v.4 no.1
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    • pp.43-56
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    • 1998
  • A multiplex PCR method was designed by employing primers specific for the eaeA gene, conserved sequences of Shiga-like toxins (SLT-I.II), and the 60-MDa plasmid of enterohemorrhagic E. coli (EHEC) O157:H7 strain. A set of six synthetic oligonucleotide primers derived from sequences of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 were used in a multiplex PCR amplification procedure to detect these genes in the same enteric pathogens. In two enterohemorrhagic E. coli O157:H7 (ATCC 35150, ATCC 43894) reference strains, PCR products of 317bps (eaeA), 228bps (SLT-I.II), and 167bps (60-MDa plasmid) were successfully amplified simultaneously in a single reaction. However, the specific PCR products were not amplified in control strains of other enteric bacteria. The sensitivity of the multiplex PCR assay for detection of the SLT-I.II, eaeA, and 60-MDa plasmid genes of E. coli O157:H7 was found to be 2.5$\times$10$^{6}$ of bacteria in diarrheal stool to amplify all three bands. The multiplex PCR technology will allow large-scale screening of many clinical specimens or contaminated foods, and will be a very useful method for the detection of a wide range of microorganisms present in the environment, including EHEC O157:H7 in various types of specimens. The multiplex PCR assay has the potential to be used as a specific and rapid method for clinical diagnosis of disease caused by EHEC O157:H7.

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Simulation of DNA/DNA Hybridization Chain Reaction Using Thermodynamic Data (열역학적 데이터에 기반한 DNA/DNA 연쇄 결합 반응 시뮬레이션)

  • 장하영;신수용;장병탁
    • Proceedings of the Korean Information Science Society Conference
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    • 2003.10b
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    • pp.772-774
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    • 2003
  • DNA/DNA의 연쇄 결합 반응에 대한 시뮬레이션을 열역학적 데이터를 이용하여 구현하였다. 1-Base의 non Watson-Crick 결합과, dangling end(결합이 이루어진 두개의 DNA strand 중 한쪽 끝이 다른 쪽 끝보다 길거나 짧은 경우)를 허용하는 nearest-neighbor model을 사용하여 구현된 이 모델에서는 한번의 hybridization만을 예측하는 것이 아니라 연속적인 결합 반응의 시뮬레이션이 가능하다. 이를 통해서 분자 알고리즘의 설계와 검증이 가능할 뿐만 아니라, cross-homology의 검사를 통한 시퀀스의 검증까지도 가능하다. 이러한 in silico 에서의 접근 방식은 효율적인 분자 알고리즘의 개발과 신뢰성 있는 시퀀스의 설계에 도움이 될 수 있다.

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PCR-Based Sensitive Detection and Identification of Xanthomonas oryzae pv. oryzae (중합효소연쇄 반응에 의한 벼 흰잎마름병균의 특이적 검출)

  • Lee, Byoung-Moo;Park, Young-Jin;Park, Dong-Suk;Kim, Jeong-Gu;Kang, Hee-Wan;Noh, Tae-Hwan;Lee, Gil-Bok;Ahn, Joung-Kuk
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.256-264
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    • 2004
  • A new primer set was developed for the detection and identification of Xanthomonas oryzae pv. oryzae, the bacterial leaf blight (BLB) pathogen in rice plant. The nucleotide sequence of hpaA gene was determined from X. o. pv. oryzae str. KACC10331, and the sequence information was used to design primers for the application of the polymerase chain reaction (PCR). The nucleotide sequence of hpaA from X. o. pv. oryzae str. KACC 10331 was aligned with those of X. campestris pv. vesicatoria, X. campestris pv. campestris, X. axonopodis pv. citri, and X. axonopodis pv. glycines. Based on these results, a primer set(XOF and XOR) was designed for the specific detection of hpaA in X. o. pv. oryzae. The length of PCR products amplified using the primer set was 534-bp. The PCR product was detected from only X. o. pv. oryzae among other Xanthomonas strains and reference bacteria. This product was used to confirm the conservation of hpaA among Xanthomonas strains by Southern-blotting. Furthermore, PCR amplification with XOF and XOR was used to detect the pathogen in an artificially infected leaf. The sensitivity of PCR detection in the pure culture suspension was also determined. This PCR-based detection methods will be a useful method for the detection and identification of X. o. pv. oryzae as well as disease forecasting.