Continuous polymerase chain reaction microfluidics integrated with a gold-capped nanoslit sensing chip for Epstein-Barr virus detection

Han Yun Hsieh, Ray Chang, Yung Yu Huang, Po Han Juan, Hidetoshi Tahara, Kuan Yi Lee, Di Ngoc Kha Vo, Ming Han Tsai, Pei Kuen Wei, Horn Jiunn Sheen, Yu Jui Fan

Research output: Contribution to journalArticlepeer-review

Abstract

We present the first combination of a microfluidic polymerase chain reaction (PCR) with a gold nanoslit-based surface plasmon resonance (SPR) sensor for detecting the DNA sequence of latent membrane protein 1 (LMP1). The PCR microchannel was produced through a laser scribing technique, and the SPR nanoslit chip was manufactured via hot-embossing nanoimprinting lithography. Afterward, the LMP1 DNA probe was adsorbed onto the SPR chip of the integrated device through electrostatic interactions for further detection. The device can complete the analytical procedure in around 36 min, while the traditional machine requires 105 min to achieve similar signals under the same PCR thermal cycles. The calibration curve with serially diluted LMP1 DNA exhibited the accuracy (R2 > 0.99) and sensitivity (limit of detection: ∼10−11 g/mL) of the device. Moreover, extracted DNA from Epstein-Barr virus (EBV)-positive cells were directly detected through the integrated chip. In brief, this all-in-one chip can amplify gene fragments at the front-end and detect them at the back-end, decreasing the time required for the analysis without compromising accuracy or sensitivity. We believe this label-free, real-time, low-cost device has enormous potential for rapid detection of various viruses, such as EBV and COVID-19.

Original languageEnglish
Article number113672
JournalBiosensors and Bioelectronics
Volume195
DOIs
Publication statusPublished - Jan 1 2022

Keywords

  • Epstein-barr virus
  • LMP1
  • Nanoslit
  • Polymerase chain reaction
  • Surface plasmon resonance

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

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