• Title/Summary/Keyword: One-step nucleic acid amplification

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What Is the Problem in Clinical Application of Sentinel Node Concept to Gastric Cancer Surgery?

  • Miyashiro, Isao
    • Journal of Gastric Cancer
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    • v.12 no.1
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    • pp.7-12
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    • 2012
  • More than ten years have passed since the sentinel node (SN) concept for gastric cancer surgery was first discussed. Less invasive modified surgical approaches based on the SN concept have already been put into practice for malignant melanoma and breast cancer, however the SN concept is not yet placed in a standard position in gastric cancer surgery even after two multi-institutional prospective clinical trials, the Japan Clinical Oncology Group trial (JCOG0302) and the Japanese Society for Sentinel Node Navigation Surgery (SNNS) trial. What is the problem in the clinical application of the SN concept to gastric cancer surgery? There is no doubt that we need reliable indicator(s) to determine with certainty the absence of metastasis in the lymph nodes in order to avoid unnecessary lymphadenectomy. There are several matters of debate in performing the actual procedure, such as the type of tracer, the site of injection, how to detect and harvest, how to detect metastases of SNs, and learning period. These issues have to be addressed further to establish the most suitable procedure. Novel technologies such as indocyanine green (ICG) fluorescence imaging and one-step nucleic acid amplification (OSNA) may overcome the current difficulties. Once we know what the problems are and how to tackle them, we can pursue the goal.

Comparison of the Real-Time Nucleic Acid Sequence-Based Amplification (NASBA) Assay, Reverse Transcription-PCR (RT-PCR) and Virus Isolation for the Detection of Enterovirus RNA. (엔테로바이러스 검출을 위한 real-time nucleic acid sequence-based amplification (NASBA), reverse transcription-PCR (RT-PCR) 및 바이러스 배양법의 비교)

  • Na, Young-Ran;Joe, Hyeon-Cheol;Lee, Young-Suk;Bin, Jae-Hun;Cheigh, Hong-Sik;Min, Sang-Kee
    • Journal of Life Science
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    • v.18 no.3
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    • pp.374-380
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    • 2008
  • Rapid detection of enterovirus (EVs) is important in the management of aseptic meningitis. We examined the relative efficiency and specificity of the real-time nucleic acid sequence-based amplification (NASBA) comparing with the established reverse transcription polymerase chain reaction (RT-PCR) and viral culture method which were used for the detection of enterovirus RNA in clinical specimens. Of the total 292 samples, 145 were found to be positive to enterovirus RNA by real-time NASBA, 101 were positive by viral culture, and 86 were positive by RT-PCR. 147 samples and 46 samples were determined to be negative and positive by all methods respectively, but 4 samples were positive only by real-time NASBA. To compare the specificity of each method, various clinical samples which were diagnosed for herpes simplex virus (HSV)-1, HSV-2, adenovirus, mumps, and rhinovirus were applied. Except one rhinovirus sample which was false positive to enterovirus RNA by RT-PCR, the other different samples were negative to all three methods. The real-time NASBA procedure can be completed within 5 hours in contrast with 9 hours for the RT-PCR and 3-14 days for the viral culture. From this study, it was suggested that the real-time NASBA assay could be a standardized, rapid, specific, and sensitive procedure for the detection of enterovirus RNA.

Rapid Detection of Fluorescent DNA on Denaturing Polyacrylamide Gel by Using Gel Scanner (겔스캐너를 이용한 변성아크릴아마이드 겔의 형광 DNA 검출)

  • Ku Ja-Hwan;Jeong Ji-Ung;Cho Young-Chan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.spc1
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    • pp.228-230
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    • 2005
  • The denature polyacrylamide gel stain silver nitrate is used for routine nucleic acid detection. More sensitive stains, such as Vistra Green, SYBR Green are available to address a broad range of DNA applications requiring lower detection limits in polyacrylamide gel formats. Gel Scanners, laser-scanning instruments, provide sensitive fluorescence detection of DNA gel stains. We established one step fluorescent impregnation enhanced sensitivity with simple, rapid and low cost. We have applied this fluorescent staining procedure for the routine analysis of DNA profiles generated by SSR amplification.