Paper-Based Airborne Bacteria Collection and DNA Extraction Kit

Abstract

The critical risk from airborne infectious diseases, bio-weapons, and harmful bacteria is currently the highest it has ever been in human history. The requirement for monitoring airborne pathogens has gradually increased to defend against bioterrorism or prevent pandemics, especially via simple and low-cost platforms which can be applied in resource-limited settings. Here, we developed a paper-based airborne bacteria collection and DNA extraction kit suitable for simple application with minimal instruments. Airborne sample collection and DNA extraction for PCR analysis were integrated in the paper kit. We created an easy-to-use paper-based air monitoring system using 3D printing technology combined with an air pump. The operation time of the entire process, comprising air sampling, bacterial cell lysis, purification and concentration of DNA, and elution of the DNA analyte, was within 20 min. All the investigations and optimum settings were tested in a custom-designed closed cabinet system. In the fabricated cabinet system, the paper kit operated effectively at a temperature of 25–35 °C and 30–70% relative humidity for air containing 10–106 CFU Staphylococcus aureus. This paper kit could be applied for simple, rapid, and cost-effective airborne pathogen monitoring.

Paper-based sensors for bacteria detection

“…Collection can be achieved by sedimentation, filtration, centrifugation, impaction, impingement or microfluidic chips 154 . For example, air samples with airborne bacteria can be passed through a porous sampling pad on a paper-based device 155 , 156 . The collected bacteria are then lysed using a dried lysis buffer on the sampling pad, releasing DNA molecules, which are then transported through water-mediated lateral flow to a binding pad.…”

Chemical Trends in Sample Preparation for Nucleic Acid Amplification Testing (NAAT): A Review

“…Washing buffers typically resemble the chemical composition of binding buffers and may contain chaotropic salts at reduced concentration to maintain binding, but they may also include alcohols (e.g., ethanol, isopropanol). Washing buffers used in laboratory-based NA extraction kits using silanol surfaces typically contain ethanol (50-100%) [51,85,124] or isopropanol (15-80%) [34,42,45] to remove salts and contaminants while precipitating the NAs [125]. Ethanol and isopropanol have a lower dielectric constant (ε) than water (ε of 24.6 and 18.3, respectively, vs. 78.5 [78]) [126,127], making the nucleic acids less hydrophilic by decreasing ionisation.…”

Low-cost microfluidics: Towards affordable environmental monitoring and assessment

“…(C) Detection of airborne bacteria with μPAD and a 3D printed device. Reproduced with permission from (Seok et al, 2021).…”

“…The device showed selectivity for levoglucosan with variation in colorimetric signal intensity lower than 8% (Dias et al, 2019). Seok et al developed a μPAD combined with a 3D printed analysis kit for detection of airborne bacteria by collecting aerosols (Seok et al, 2021) (Figure 6C).…”