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Nucleic Acids Research 2005 33(18):e156; doi:10.1093/nar/gni157
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Published online 12 October 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
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Methods Online

Miniature RT–PCR system for diagnosis of RNA-based viruses

Chia-Sheng Liao1, Gwo-Bin Lee1,2,*, Hsiao-Sheng Liu3, Tsung-Min Hsieh4 and Ching-Hsing Luo4

1Institute of MEMS Engineering, National Cheng Kung University Tainan, Taiwan 701, The Republic of China 2Department of Engineering Science, National Cheng Kung University Tainan, Taiwan 701, The Republic of China 3Department of Microbiology and Immunology, National Cheng Kung University Tainan, Taiwan 701, The Republic of China 4Department of Electrical Engineering, National Cheng Kung University Tainan, Taiwan 701, The Republic of China

*To whom correspondence should be addressed. Tel: +886 6 2757575 Ext. 63347; Fax: +886 6 2761687; Email: gwobin{at}mail.ncku.edu.tw

Received March 31, 2005. Revised July 23, 2005. Accepted September 22, 2005.

This paper presents an innovative portable chip-based RT–PCR system for amplification of specific nucleic acid and detection of RNA-based viruses. The miniature RT–PCR chip is fabricated using MEMS (Micro-electro-mechanical-system) techniques, and comprises a micro temperature control module and a PDMS (polydimethylsiloxane)-based microfluidic control module. The heating and sensing elements of temperature control module are both made of platinum and are located within the reaction chambers in order to generate a rapid and uniform thermal cycling. The microfluidic control module is capable of automating testing process with minimum human intervention. In this paper, the proposed miniature RT–PCR system is used to amplify and detect two RNA-based viruses, namely dengue virus type-2 and enterovirus 71 (EV 71). The experimental data confirm the ability of the system to perform a two-step RT–PCR process. The developed miniature system provides a crucial tool for the diagnosis of RNA-based viruses.


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