Label-free electrical sensing of bacteria in eye wash samples: A step towards point-of-care detection of pathogens in patients with infectious keratitis.

Hardik J Pandya, Manoj Kumar Kanakasabapathy, Saloni Verma, Manjyot Kaur Chug, Adnan Memic, Mihaela Gadjeva, Hadi Shafiee
Author Information
  1. Hardik J Pandya: Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital - Harvard Medical School, Boston, MA 02115, USA.
  2. Manoj Kumar Kanakasabapathy: Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital - Harvard Medical School, Boston, MA 02115, USA.
  3. Saloni Verma: Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital - Harvard Medical School, Boston, MA 02115, USA.
  4. Manjyot Kaur Chug: Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital - Harvard Medical School, Boston, MA 02115, USA.
  5. Adnan Memic: Center for Nanotechnology, King AbdulAziz University, Jeddah 21589, Saudi Arabia.
  6. Mihaela Gadjeva: Division of Infectious Disease, Department of Medicine, Brigham and Women's Hospital - Harvard Medical School, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, 25 Shattuck Street, Boston, MA 02115, USA.
  7. Hadi Shafiee: Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital - Harvard Medical School, Boston, MA 02115, USA; Department of Medicine, Harvard Medical School, 25 Shattuck Street, Boston, MA 02115, USA. Electronic address: HSHAFIEE@BWH.HARVARD.EDU.

Abstract

The diagnosis of keratitis is based on visual exam, tissue cytology, and standard microbial culturing to determine the type of the infectious pathogen. To prescribe appropriate therapy, it is important to distinguish between bacterial, fungal, and viral keratitis, as the treatments are quite different. Diagnosis of the causative organism has a substantial prognostic importance. Further, timely knowledge of the nature of the pathogen is also critical to adapt therapy in patients unresponsive to empiric treatment options, which occurs in 10% of all cases. Currently, the identification of the nature of the pathogen that causes keratitis is achieved via microbial culture screening, which is laboratory-based, expensive, and time-consuming. The most frequent pathogens that cause the corneal ulcers are P. aeruginosa and S. aureus. Here, we report a microchip for rapid (<1h) detection of P. aeruginosa (6294), S. aureus(LAC), through on-chip electrical sensing of bacterial lysate. We evaluated the microchip with spiked samples of PBS with bacteria concentration between 10 to 10 CFU/mL. The least diluted bacteria concentration in bacteria-spiked samples with statistically significant impedance change was 10 CFU/mL. We further validated our assay by comparing our microchip results with the standard culture-based methods using eye washes obtained from 13 infected mice.

Keywords

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Grants

  1. R01 AI118502/NIAID NIH HHS
  2. R01 EY022054/NEI NIH HHS

MeSH Term

Animals
Biosensing Techniques
Electric Impedance
Equipment Design
Humans
Keratitis
Lab-On-A-Chip Devices
Limit of Detection
Mice
Mice, Inbred C57BL
Point-of-Care Systems
Pseudomonas Infections
Pseudomonas aeruginosa
Staphylococcal Infections
Staphylococcus aureus
Tears

Word Cloud

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