4.7 Article

PCR-coupled Paper-based Surface-enhanced Raman Scattering (SERS) Sensor for Rapid and Sensitive Detection of Respiratory Bacterial DNA

期刊

SENSORS AND ACTUATORS B-CHEMICAL
卷 326, 期 -, 页码 -

出版社

ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2020.128802

关键词

Bacterial DNA detection; Surface Enhanced Raman Scattering; Point-of-Care test (POCT); Molecular Diagnostics

资金

  1. Fundamental Research Program of the Korean Institute of Materials Science (KIMS) [PNK 6800, PNK 6970]
  2. Nano Product Upgrading Program using Electron Beam through the Gyeongsangnamdo Gimhae

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A paper-based surface-enhanced Raman scattering (SERS) substrate was developed for rapid detection of respiratory bacterial DNA, coupled with polymerase chain reaction (PCR). The PCR-coupled SERS method showed enhanced detection capability after 10 amplification cycles, enabling low-cycle amplification and sensitive DNA sensing.
A paper-based surface-enhanced Raman scattering (SERS) substrate composed of silver-nanowires (AgNWs) was coupled with polymerase chain reaction (PCR) for rapid and sensitive determination of respiratory bacterial DNA. EvaGreen dye, which is a DNA intercalating molecule, was introduced to the low-cycled PCR product, and the difference in dye intercalation in the DNA structure was quantified by Raman spectroscopy to verify the presence of a target gene. Mycoplasma pneumoniae (M. Pne), was selected as a target for the sensing demonstration. At various gene concentrations and amplification cycles, detection capability of fluorescence-based realtime PCR (RT-PCR) and the PCR-coupled SERS method were compared to develop a DNA detection system capable of low-cycle amplification and sensitive DNA sensing. After 10 cycles, the PCR-coupled SERS method showed enhanced detection capability with a DNA detection limit of 3.12 pg/mu L. Then, the SERS substrate was prepared as a rapid kit that included a test line and negative and positive control lines. Target DNA after 10 cycles of amplification was successfully discriminated compared to non-target DNA and it was statistically relevant. The developed system is expected to be used as a detection method for various bacteria and viruses, and also be integrated with diverse POC diagnostic systems.

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