Title page for etd-0811117-060846


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URN etd-0811117-060846
Author Hui-wen Huang
Author's Email Address No Public.
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Department Biological Sciences
Year 2016
Semester 2
Degree Master
Type of Document
Language zh-TW.Big5 Chinese
Title Using M-class Ultra-Pure Liquid Chromatography/ Mass Spectrometry (UPLC/MSE) for Rapid Detection and Typing of Norovirus Capsid Protein
Date of Defense 2017-07-27
Page Count 69
Keyword
  • M-class ultra-performance liquid chromatography
  • mass spectrometry
  • matrix-assisted laser desorption ionization
  • Norovirus
  • virus-like particle
  • Abstract Norovirus is the leading etiologic pathogen of diarrheal disease worldwide. By meta-analysis, norovirus was associated with 212,000 deaths annually, the disease burden of norovirus is high. Noroviruses are non-enveloped, single-stranded RNA viruses classified as genus Norovirus of the family Caliciviridae. The norovirus capsid is composed by VP1 proteins with shell (S) and protruding (P1 and P2) domains, in the order of S-P1-P2-P1 from N to C-terminus. The most variable P2 subdomain is insertion within the P1 domain. Based on the VP1 sequence, norovirus were divided into genogroups GI–GVII. GI, GII, and GIV are known to be the human pathogens, with GII being the most common in massive outbreaks. Due to norovirus are uncultivable in clinical laboratory, the combined reverse transcription polymerase chain reaction (RT- PCR) methods directly detect norovirus from specimens are widely used. However, the highly sensitive amplification might lead to false-positive results, while the high mutation rates of RNA viruses might lead to a negative result in the sequence dependent amplification. A GII.4 2006b norovirus like particle (VLP) was used to analysis for the signature signal of peptide profile. The matrix-assisted laser desorption ionization (MALDI) MS has used for examine the molecular weight of VLP. Total proteins from VLP were digested with trypsin, and separated on an M-class ultra-performance liquid chromatography (UPLC). Parallel ion fragmentation was programmed and signals were collected from 50 to 1500 m/z. Data was processed with ProgenesisTM QI-P for identification and quantification. Thus, the results in this study reveal that combination with more sensitive and rapid method for M-class UPLC and Xevo® G2 Q-TOF / MS has the potential to replace current detection methods. Keywords: Norovirus; virus-like particle; M-class ultra-performance liquid chromatography; mass spectrometry; matrix-assisted laser desorption ionization
    Advisory Committee
  • Chuan-Fa Chang - chair
  • Chung-Ren Lin - co-chair
  • Liang-Yin Ke - advisor
  • Jiin-Tsuey Cheng - advisor
  • Files
  • etd-0811117-060846.pdf
  • Indicate in-campus at 5 year and off-campus access at 5 year.
    Date of Submission 2017-09-11

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