Continuous polymerase chain reaction microfluidics integrated with a gold-capped nanoslit sensing chip for Epstein-Barr virus detection
Journal
Biosensors and Bioelectronics
Journal Volume
195
Date Issued
2022
Author(s)
Sheen H.-J
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. ? 2021
Subjects
Epstein-barr virus
LMP1
Nanoslit
Polymerase chain reaction
Surface plasmon resonance
DNA
Gold
Microfluidics
Nanoimprint lithography
Nanostructures
Surface reactions
Viruses
Epstein-Barr virus
Hot-embossing
Laser scribing
Latent membrane protein 1
Membrane proteins
Nanoslits
Sensing chip
Surface plasmon resonance sensor
Surface-plasmon resonance
Virus detection
gold nanoparticle
latent membrane protein 1
gold
matrix protein
Article
calibration
controlled study
DNA extraction
DNA sequence
Epstein Barr virus
limit of detection
measurement accuracy
microfluidics
nonhuman
polymerase chain reaction
sensitivity analysis
static electricity
surface plasmon resonance
virus detection
Epstein Barr virus infection
genetic procedures
genetics
human
Biosensing Techniques
COVID-19
Epstein-Barr Virus Infections
Herpesvirus 4, Human
Humans
Polymerase Chain Reaction
SARS-CoV-2
Viral Matrix Proteins
Type
journal article