Micromachined polymerase chain reaction system for multiple DNA amplification of upper respiratory tract infectious diseases.
This paper presents a micro polymerase chain reaction (PCR) chip for the DNA-based diagnosis of microorganism genes and the detection of their corresponding antibiotic-resistant genes. The micro PCR chip comprises cheap biocompatible soda-lime glass substrates with integrated thin-film platinum resistors as heating/sensing elements, and is fabricated using micro-electro-mechanical-system (MEMS) techniques in a reliable batch-fabrication process. The heating and temperature sensing elements are made of the same material and are located inside the reaction chamber in order to ensure a uniform temperature distribution. This study performs the detection of several genes associated with upper respiratory tract infection microorganisms, i.e. Streptococcus pneumoniae, Haemopilus influenze, Staphylococcu aureus, Streptococcus pyogenes, and Neisseria meningitides, together with their corresponding antibiotic-resistant genes. The lower thermal inertia of the proposed micro PCR chip relative to conventional bench-top PCR systems enables a more rapid detection operation with reduced sample and reagent consumption. The experimental data reveal that the high heating and cooling rates of the system (20 and 10 degrees C/s, respectively) permit successful DNA amplification within 15 min. The micro PCR chip is also capable of performing multiple DNA amplification, i.e. the simultaneous duplication of multiple genes under different conditions in separate reaction wells. Compared with the large-scale PCR system, it is greatly advantageous for fast diagnosis of multiple infectious diseases. Multiplex PCR amplification of two DNA segments in the same well is also feasible using the proposed micro device. The developed micro PCR chip provides a crucial tool for genetic analysis, molecular biology, infectious disease detection, and many other biomedical applications.
Liao
Chia-Sheng
CS
Institute of Micro-Electro-Mechanical-System Engineering, National Cheng Kung University, Tainan 701, Taiwan.
Lee
Gwo-Bin
GB
Wu
Jiunn-Jong
JJ
Chang
Chih-Ching
CC
Hsieh
Tsung-Min
TM
Huang
Fu-Chun
FC
Luo
Ching-Hsing
CH
D004586:Electrophoresis; D006358:Hot Temperature; D016133:Polymerase Chain Reaction; D012141:Respiratory Tract Infections / Q000378:metabolism*; D013997:Time Factors