4.7 Article

All-in-One DNA Extraction Tube for Facilitated Real-Time Detection of Infectious Pathogens

Journal

ADVANCED HEALTHCARE MATERIALS
Volume 10, Issue 14, Pages -

Publisher

WILEY
DOI: 10.1002/adhm.202100430

Keywords

all-in-one tubes; nucleic acid extraction; pathogen detection; real-time polymerase chain reaction; tertiary-amine-containing functional polymers

Funding

  1. Ministry of Science and ICT (MSIT, Korea) - Ministry of Trade, Industry & Energy (MOTIE, Korea) [2017R1A2B3007806, 20008777, P0002007]
  2. Nano Open Innovation Lab Cooperation Project of National NanoFab Center (NNFC) in 2020

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The all-in-one tube platform developed allows genetic analysis involving DNA extraction, amplification, and detection to be performed in a single tube with high efficiency and compatibility with existing real-time PCR instruments. It provides high sensitivity, selectivity, and reliability for accurate detection of various pathogens and can be used in food safety, forensic, and clinical diagnostic applications.
y An all-in-one tube platform is developed, where the genetic analysis involving DNA extraction, amplification, and detection can be performed in a single tube. The all-in-one tube consists of a polymerase chain reaction (PCR) tube in which the inner surface is conformally modified with a tertiary-amine-containing polymer to generate a strong electrostatic interaction with DNA. The all-in-one tube provides high DNA capture efficiency exceeding 80% from Escherichia coli O157: H7 pathogen at a wide range of DNA amount from 0.003 to 3 ng. Indeed, the use of the surface-functionalized PCR tube enables direct amplification and detection of the surface-captured DNA without the modification of standard real-time PCR instrument. Besides, this platform has sensitivity, selectivity, and reliability enough for accurate detection at the minimal infective dose of both gram-positive and negative pathogens. The all-in-one tube enables the direct molecular diagnosis, substantially reducing the labor-intensive pathogen detection steps while providing high compatibility with the currently established real-time PCR instruments, and illustrates its on-site applicability with convenience expandable to various genetic analyses including food safety testing, forensic analysis, and clinical diagnosis.

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