Dataset Information

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

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 101 to 108 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.

SUBMITTER: Pandya HJ

PROVIDER: S-EPMC5323318 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature

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

Pandya Hardik J HJ Kanakasabapathy Manoj Kumar MK Verma Saloni S Chug Manjyot Kaur MK Memic Adnan A Gadjeva Mihaela M Shafiee Hadi H

Biosensors & bioelectronics 20161213

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 e ...[more]

PMID: 27987408

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Project description:Infectious keratitis is a major global cause of visual impairment and blindness, often affecting marginalized populations. Proper diagnosis of the causative organism is critical, and although culture remains the prevailing diagnostic tool, newer techniques such as in vivo confocal microscopy are helpful for diagnosing fungus and Acanthamoeba. Next-generation sequencing holds the potential for early and accurate diagnosis even for organisms that are difficult to culture by conventional methods. Topical antibiotics remain the best treatment for bacterial keratitis, and a recent review found all commonly prescribed topical antibiotics to be equally effective. However, outcomes remain poor secondary to corneal melting, scarring, and perforation. Adjuvant therapies aimed at reducing the immune response associated with keratitis include topical corticosteroids. The large, randomized, controlled Steroids for Corneal Ulcers Trial found that although steroids provided no significant improvement overall, they did seem beneficial for ulcers that were central, deep or large, non-Nocardia, or classically invasive Pseudomonas aeruginosa; for patients with low baseline vision; and when started early after the initiation of antibiotics. Fungal ulcers often have worse clinical outcomes than bacterial ulcers, with no new treatments since the 1960s when topical natamycin was introduced. The randomized controlled Mycotic Ulcer Treatment Trial (MUTT) I showed a benefit of topical natamycin over topical voriconazole for fungal ulcers, particularly among those caused by Fusarium. MUTT II showed that oral voriconazole did not improve outcomes overall, although there may have been some effect among Fusarium ulcers. Given an increase in nonserious adverse events, the authors concluded that they could not recommend oral voriconazole. Viral keratitis differs from bacterial and fungal cases in that it is often recurrent and is common in developed countries. The Herpetic Eye Disease Study (HEDS) I showed a significant benefit of topical corticosteroids and oral acyclovir for stromal keratitis. HEDS II showed that oral acyclovir decreased the recurrence of any type of herpes simplex virus keratitis by approximately half. Future strategies to reduce the morbidity associated with infectious keratitis are likely to be multidimensional, with adjuvant therapies aimed at modifying the immune response to infection holding the greatest potential to improve clinical outcomes.

Project description:The microbiological profile of infectious keratitis has shown great differences across the world. Due to the continuous shifting trends in microbiological profile and antibiotic resistance patterns reported in several studies, constant local updates are crucial to provide an adequate treatment. The propose of this study was to analyze the incidence of infectious keratitis, possible changing trends in microbiological profile, and bacteria sensitivity to commonly used antibiotics, in our tertiary center, in the last 10 years. A retrospective study was performed, based on the survey review of electronic medical records of all patients with presumed infectious keratitis, between January 1, 2009, and December 31, 2018. Microbial cultures were performed, and patients were treated according to an internal protocol. A total of 1360 samples were included. We obtained a 35.1% culture-positive rate. Bacteria accounted for 76.78% of all positive scrapes (53.34% were Gram positive and 23.44% were Gram negative), Acanthamoeba for 12.13%, fungi for 8.16%, and virus for 2.93%. The most frequent agent identified was Corynebacterium macginleyi (18.41%), followed by Staphylococcus aureus (17.78%), Streptococcus pneumoniae (9.41%), and Pseudomonas aeruginosa (9.00%). We identified at least one ophthalmologic risk factor in 410 patients (85.77%). Trauma and contact lens wear were the most common risk factors found, accounting for 34.94% (n?=?167) and 33.47% (n?=?160) of cases. Sensitivity to fluoroquinolones and aminoglycosides was tested in all bacterial isolates, presenting values of 96.66% and 98.12%. In our region, the most common bacteria are Staphylococcus aureus, Streptococcus pneumoniae, and Pseudomonas aeruginosa, and they showed high sensitivity rates to first-line antibiotics, without any modification or emergence of antibiotic resistance trends during the 10 years of the study. For this reason, we decided to maintain the same internal protocol in our tertiary centre.

Dataset Information

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

Publications

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

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