Point-of-care testing system for digital single cell detection of MRSA directly from nasal swabs
| Autor: | Roland Zengerle, Michael Jehle, Mara Specht, Florian Hausladen, Simon Wadle, Silvia Calabrese, Ulrike Goetz, Nadine Borst, Anna M. Sobieraj, Holger Wurm, Annerose Serr, Martin J. Loessner, Anja Gerhardts, Philip Koch, Raimund Rother, Karl Stock, Mathias Schmelcher, Felix von Stetten, Martin Schulz, Dominik Drossart, Nils Paust, Jia Li, Philipp Tepper, Martin Meyer, Marina Handel, Georg Haecker, Fritz Eichenseher |
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| Rok vydání: | 2020 |
| Předmět: |
Methicillin-Resistant Staphylococcus aureus
Staphylococcus aureus Point-of-care testing Biomedical Engineering Recombinase Polymerase Amplification Bioengineering 02 engineering and technology medicine.disease_cause 01 natural sciences Biochemistry Rapid detection Microbiology Bacterial Proteins medicine Humans Detection limit business.industry 010401 analytical chemistry General Chemistry Staphylococcal Infections 021001 nanoscience & nanotechnology 0104 chemical sciences Real-time polymerase chain reaction Point-of-Care Testing Nasal Swab 0210 nano-technology business Staphylococcus |
| Zdroj: | Lab on a Chip. 20:2549-2561 |
| ISSN: | 1473-0189 1473-0197 |
| Popis: | We present an automated point-of-care testing (POCT) system for rapid detection of species- and resistance markers in methicillin-resistant Staphylococcus aureus (MRSA) at the level of single cells, directly from nasal swab samples. Our novel system allows clear differentiation between MRSA, methicillin-sensitive S. aureus (MSSA) and methicillin-resistant coagulase-negative staphylococci (MR-CoNS), which is not the case for currently used real-time quantitative PCR based systems. On top, the novel approach outcompetes the culture-based methods in terms of its short time-to-result (1 h vs. up to 60 h) and reduces manual labor. The walk-away test is fully automated on the centrifugal microfluidic LabDisk platform. The LabDisk cartridge comprises the unit operations swab-uptake, reagent pre-storage, distribution of the sample into 20 000 droplets, specific enzymatic lysis of Staphylococcus spp. and recombinase polymerase amplification (RPA) of species (vicK) - and resistance (mecA) -markers. LabDisk actuation, incubation and multi-channel fluorescence detection is demonstrated with a clinical isolate and spiked nasal swab samples down to a limit of detection (LOD) of 3 ± 0.3 CFU μl-1 for MRSA. The novel approach of the digital single cell detection is suggested to improve hospital admission screening, timely decision making, and goal-oriented antibiotic therapy. The implementation of a higher degree of multiplexing is required to translate the results into clinical practice. |
| Databáze: | OpenAIRE |
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