• Title/Summary/Keyword: polymerase chain reaction

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A Case of Type 1 Herpes Simplex Virus Encephalitis Detected by Polymerase Chain Reaction (중합효소연쇄반응으로 확진된 Herpes Simplex virus 뇌염 1례)

  • Park, Dae Young;Lee, Joon Soo;Lee, Young Ho;Sohn, Young Mo
    • Pediatric Infection and Vaccine
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    • v.3 no.2
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    • pp.207-213
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    • 1996
  • Herpes simplex virus(HSV) infections of the CNS are associated with significant morbidity and mortality even when appropriate antiviral therapy is administered. HSV infections of the brain can be subdivided into two categories : neonatal HSV infections, which usually are caused by HSV type 2, and herpes simplex encephalitis(HSE), which occur in patients over 3 months old and is nearly uniformly caused by HSV type 1. The clinical presentation of HSE is one of the focal encephalopathic process associated with altered levels of consciousness, fever, focal seizures and hemiparesis. But because of the lack of pathognomic clinical presentation and diagnostic procedure, the efforts to develop alternative diagnostic procedure have led to the use of new diagnostic technique such as polymerase chain reaction(PCR). We report a case of HSV type 1 encephalitis in 13 month old male infant who presented with altered level of consciousness, fever and focal seizures. With the use of the PCR, HSV-1 DNA was detected in cerebrospinal fluid from the patient. The symptoms and signs of encephalitis subsided by treatment with acyclovir in 14 days.

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A STUDY ON MUTATIONS OF P53 TUMOR SUPPRESSOR GENE IN ORAL TUMORS (구강종양에서 p53 종양억제 유전자의 돌연변이에 관한 연구)

  • Joo, Seong-Chai;Pyo, Sung-Woon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.1
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    • pp.45-52
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    • 2000
  • Nowadays, there are a lot of evidence that mutation of the p53 tumor suppressor gene is one of the most common genetic abnormalities in neoplastic progression. In this study, we analyzed 20 specimens of oral tumors(squamous cell carcinoma 14 cases, ameloblastoma 3 cases, adenoid cystic carcinoma 2 cases, malignant schwannoma 1 case)using polymerase chain reaction and direct sequencing which used an automated DNA sequencer and software for detection of mutations. Polymerase chain reactions were performed with 4 sets of primers encompassing exon 5, 6, 7, 8, and direct sequencing method was employed. The results were as followings. 1. We detected 10 point mutations out of 20 specimens (50%). 2. The genetic alterations included 7 mis-sense mutations resulting in single amino acid subtitutions, 2 silent mutations, 1 non-sense mutations encoding a stop codon. 3. Mutations were mostly in exon 7(7 out of 10 mutations, 70%) and involved codons 225, 234, 235, 236, 238, 247. 4. Therse were 4 cases of $T{\rightarrow}A$ transversion, 2 cases of $C{\rightarrow}A$ transversion, $A{\rightarrow}G$ transition, 1 case of $C{\rightarrow}G$, $T{\rightarrow}G$ transversion respectively. 5. We could find out point mutations more conveniently using PCR - Automated Direct Sequencing method.

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A Simple Detection of Sweetpotato Feathery Mottle Virus by Reverse Transcription Polymerase Chain Reaction

  • Jeong Jae-Hun;Chakrabarty Debasis;Kim Young-Seon;Eun Jong-Seon;Choi Yong-Eui;Paek Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.5 no.2
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    • pp.83-86
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    • 2003
  • A reverse transcription polymerase chain reaction (RT-PCR) protocol was developed using two specific 22-mer primers located in coat protein gene of SPFMV. A 411 bp PCR-product was detected in virus infected plants as well as tissue culture raised sweet potato but not in healthy plants. For optimization of RT-PCR protocol, the optimum crude nucleic acid concentration, annealing temperature, primer concentration and numbers of PCR-cycle for maximum sensitivity and specificity were determined. The optimum condition for RT-PCR was as follows: RT-PCR reaction mixture was one-step mixture, containing 50 pmol of primer, 30 units of reverse transcriptase, 5 units of RNasin, and the crude nucleic acid extracts (200 ng). In RT-PCR, cDNA was synthesized at $42^{\circ}C$ for 45 min before a quick incubation on ice after pre-denaturation at $95^{\circ}C$ for 5 min. The PCR reaction was carried out for 40 cycles at $96^{\circ}C$ for 30 see, $63^{\circ}C$ for 30 sec, $72^{\circ}C$ for 1 min, and finally at $72^{\circ}C$ for 10 min. The viral origin of the amplified product was confirmed by sequencing, with the sequence obtained having $95-98\%$ homology with published sequence data for SPFMV. The benefits of this RT-PCR based detection of SPFMV would be simple, rapid and specific.

Detection of DNA Sequence Polymorphism by Polymerase Chain Reaction in Fraxinus mandshurica Rupr Growing in Korea (P.C.R 기법(技法)을 이용(利用)한 들메나무 DNA sequence의 변이조사(變異調査))

  • Na, Chun Soo;Noh, Eun Woon;Kim, Young Joong;Shin, Chang Ho;Song, Won Sup;Kim, Sea Hyun
    • Journal of Korean Society of Forest Science
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    • v.81 no.4
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    • pp.320-324
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    • 1992
  • It has been reported that there are two distinct phenotypes in Fraxinus mandshurica Rupr. growing in Korea. Recently developed polymerase chain reaction(PCR) was used to detect DNA sequence polymorphism in the species. Using a thermostable DNA polymerase and synthetic DNA primers, unknown DNA sequences from the species were randomly amplified. The two types of the species produced different DNA amplification pattern with three different primers tested. Although DNA polymorphism was detected among individuals within types, each type has its own distinct pattern. The two types could be easily differentiated by trier characteristic predominant bands.

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Slipchip Device Development in Molecular Diagnostics

  • Qingtian Yin;Huiwen Bai;Ruijie Li;Youngung Seok
    • Korean Journal of Materials Research
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    • v.34 no.2
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    • pp.63-71
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    • 2024
  • Slipchip offers advantages such as high-throughout, low cost, and simple operation, and therefore, it is one of the technologies with the greatest potential for high-throughput, single-cell, and single-molecule analyses. Slipchip devices have achieved remarkable advances over the past decades, with its simplified molecular diagnostics gaining particular attention, especially during the COVID-19 pandemic and in various infectious diseases scenarios. Medical testing based on nucleic acid amplification in the Slipchip has become a promising alternative simple and rapid diagnostic tool in field situations. Herein, we present a comprehensive review of Slipchip device advances in molecular diagnostics, highlighting its use in digital recombinase polymerase amplification (RPA), loop-mediated isothermal amplification (LAMP), and polymerase chain reaction (PCR). Slipchip technology allows users to conduct reliable droplet transfers with high-throughput potential for single-cell and molecule analyses. This review explores the device's versatility in miniaturized and rapid molecular diagnostics. A complete Slipchip device can be operated without special equipment or skilled handling, and provides high-throughput results in minimum settings. This review focuses on recent developments and Slipchip device challenges that need to be addressed for further advancements in microfluidics technology.