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KMID : 0380020090240050432
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Korean Journal of Biotechnology and Bioengineering
2009 Volume.24 No. 5 p.432 ~ p.438
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Highly Sensitive Detection of Pathogenic Bacteria Using PDMS Micro Chip Containing Glass Bead
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Won Ji-Yeong
Min Jun-Hong
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Abstract
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Here, we demonstrated simple nucleic acid, RNA, concentration method using polymer micro chip containing glass bead (). Polymer micro chip was fabricated by PDMS (, in the height) including pillar structure (, gap size ) for blocking micro bead. RNA could be adsorbed on micro glass bead at low pH by hydrogen bonding whereas RNA was released at high pH by electrostatic force between silica surface and RNA. Amount of glass beads and flow rate were optimized in aspects of adsorption and desorption of RNA. Adsorption and desorption rate was measured with real time PCR. This concentrated RNA was applied to amplification micro chip in which NASBA (Nucleic Acid Sequence Based Amplification) was performed. As a result, E.coli O157 : H7 in the concentration of 10 c.f.u./10 mL was successfully detected by these serial processes (concentration and amplification) with polymer micro chips. It implies this simple concentration method using polymer micro chip can be directly applied to ultra sensitive method to measure viable bacteria and virus in clinical samples as well as environmental samples.
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KEYWORD
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RNA, Nucleic Acid Sequence Based Amplification (NASBA), PDMS, nucleic acid concentration
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