Rapid micro-PCR system for hepatitis C virus amplification

Yu-Cheng Lin; Ming-Yuan Huang; Kung-Chia Young; Ting-Tsung Chang; Ching-Yi Wu

doi:10.1117/12.395627

15 August 2000 Rapid micro-PCR system for hepatitis C virus amplification

Yu-Cheng Lin, Ming-Yuan Huang, Kung-Chia Young, Ting-Tsung Chang, Ching-Yi Wu

Proceedings Volume 4176, Micromachined Devices and Components VI; (2000) https://doi.org/10.1117/12.395627
Event: Micromachining and Microfabrication, 2000, Santa Clara, CA, United States

Abstract

A rapid micro-polymerase chain reaction ((mu) -PCR) system was integrated to amplify the complementary DNA (cDNA) molecules of hepatitis C virus (HCV). This system consists of a rapid thermal cycling system and a (mu) PCR chip fabricated by MEMS fabrication techniques. This rapid (mu) PCR system is verified by using serum samples from patients with chronic hepatitis C. The HCV amplicon of the rapid (mu) PCR system was analyzed by slab gel electrophoresis with separation of DNA marker in parallel. The (mu) PCR chip was fabricated on silicon wafer and Pyrex glass using photolithography, wet etching, and anodic bonding methods. Using silicon material to fabricate the raction well improves the temperature uniformity of sample and helps to reach the desired temperature faster. The rapid close loop thermal cycling system comprises power supplies, a thermal generator, a computer control PID controller, and a data acquisition subsystem. The thermoelectric (T.E.) cooler is used to work as the thermal generator and a heat sink by controlling the polarity of supplied power. The (mu) PCR system was verified with traditional PCR equipment by loading the same PCR mixture with HCV cDNA and running the same cycle numbers, then comparing both HCV amplicon slab gel electrophoresis. The HCV amplicon from the (mu) PCR system shows a DNA fragment with an expected size of 145 base pairs. The background is lower with the (mu) PCR system than that with the tradional PCR equipment. Comparing the traditional PCR equipment which spends 5.5 hours for 30 cycles to gain the detectable amount of HCV amplicon in slab gel separation, this (mu) PCR system takes 30 minutes to finish the 30 thermal cycles. This work has demonstrated that this rapid (mu) PCR system can provide rapid heat generation and dissipation, improved temperature uniformity in DNA amplification.

Citation Download Citation

Yu-Cheng Lin, Ming-Yuan Huang, Kung-Chia Young, Ting-Tsung Chang, and Ching-Yi Wu "Rapid micro-PCR system for hepatitis C virus amplification", Proc. SPIE 4176, Micromachined Devices and Components VI, (15 August 2000); https://doi.org/10.1117/12.395627

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