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

Abstract

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.

Advances in deoxyribonucleic acid extraction techniques and point-of-care molecular diagnosis of foodborne pathogens

“…from the lysate. 13,[21][22][23] Significantly, it incorporates simple automation, high selectivity, and relatively inexpensive equipment. Due to its unique benefits, SPE has become a popular extraction technique and served as a model for miniaturized sample preparation methods.…”

“…87 In terms of eliminating high-end instruments and facilities, organic polymers are exclusively involved in the POC platform to detect infectious diseases. 23,33,91 Recently, a cationic copolymer whose surface efficiently captures DNA was synthesized. A customized device with an all-in-one function was designed to apply the sample-in-answer-out format of E. coli.…”

CRISPR-Cas12a Coupled with DNA Nanosheet-Amplified Fluorescence Anisotropy for Sensitive Detection of Biomolecules

“…However, the sensitivity of this DNS-amplified FA strategy needs to be further improved because a target just can cause an FA change of one fluorophore group. In some traditional biosensing methods, catalytic hairpin assembly (CHA), , hybridization chain reaction (HCR), − polymerase chain reaction (PCR), − and other cyclic amplification technologies − are usually used to improve the sensitivity. However, these cyclic amplification technologies require specialized instruments and technicians and were more complex to operate than methods that do not require cyclic amplification.…”

Polyaniline-based 3D Network Structure Promotes Entrapment and Detection of Drug-Resistant Bacteria

“…Recently, nano-topological technologies have drawn signi cant attention for recovering and diagnosing pathogenic bacteria owing to their high surface area, controllable surface characteristics, mechanical, chemical, and thermal stability, and compatibility in molecular analysis [24][25][26]. In addition, various nanostructures and chemical modi cations have been employed to entrap different types of pathogens and recover their nucleic acids via physicochemical interactions [27][28][29][30][31]. In particular, two-dimensional (2D) nanoarrays and three-dimensional (3D) hierarchical nanostructures exhibit excellent performance by introducing topographical interactions with extracellular matrices of bacteria [32][33][34].…”