Pneumatic nano-sieve for CRISPR-based detection of drug-resistant bacteria

Nanoscale Horiz. 2023 Nov 20;8(12):1677-1685. doi: 10.1039/d3nh00365e.

Abstract

The increasing prevalence of antibiotic-resistant bacterial infections, particularly methicillin-resistant Staphylococcus aureus (MRSA), presents a significant public health concern. Timely detection of MRSA is crucial to enable prompt medical intervention, limit its spread, and reduce antimicrobial resistance. Here, we introduce a miniaturized nano-sieve device featuring a pneumatically-regulated chamber for highly efficient MRSA purification from human plasma samples. By using packed magnetic beads as a filter and leveraging the deformability of the nano-sieve channel, we achieved an on-chip concentration factor of ∼15-fold for MRSA. We integrated this device with recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas detection system, resulting in an on-chip limit of detection (LOD) of approximately 100 CFU mL-1. This developed approach provides a rapid, precise, and centrifuge-free solution suitable for point-of-care diagnostics, with the potential to significantly improve patient outcomes in resource-limited medical conditions.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacteria
  • Children's Health Insurance Program*
  • Humans
  • Limit of Detection
  • Methicillin-Resistant Staphylococcus aureus* / genetics
  • Nucleotidyltransferases

Substances

  • Nucleotidyltransferases