Rapid micro-PCR system for hepatitis C virus amplification

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

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

A rapid micro-polymerase chain reaction (μ-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 μPCR chip fabricated by MEMS fabrication techniques. This rapid μPCR system is verified by using serum samples from patients with chronic hepatitis C. The HCV amplicon of the rapid μPCR system was analyzed by slab gel electrophoresis with separation of DNA marker in parallel. The μPCR chip was fabricated on silicon wafer and Pyrex glass using photolithography, wet etching, and anodic bonding methods. Using silicon material to fabricate the reaction 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 μPCR 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 in slab gel electrophoresis. The HCV amplicon from the μPCR system shows a DNA fragment with an expected size of 145 base pairs. The background is lower with the μPCR system than that with the traditional 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 μPCR system takes 30 minutes to finish the 30 thermal cycles. This work has demonstrated that this rapid μPCR system can provide rapid heat generation and dissipation, improved temperature uniformity in DNA amplification.

Original languageEnglish
Pages (from-to)188-193
Number of pages6
JournalProceedings of SPIE - The International Society for Optical Engineering
Volume4176
Issue number1
DOIs
Publication statusPublished - 2000 Aug 15

All Science Journal Classification (ASJC) codes

  • Electronic, Optical and Magnetic Materials
  • Condensed Matter Physics
  • Computer Science Applications
  • Applied Mathematics
  • Electrical and Electronic Engineering

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