Integrated DNA and RNA extraction and purification on an automated microfluidic cassette from bacterial and viral pathogens causing community-acquired lower respiratory tract infections
| Autor: | Tina Roeser, Marta Camps, Antoni Homs Corbera, Surbhi Malhotra-Kumar, Carmen Schwind, Peter Spang, Francesc Guasch, Herman Goossens, Benjamin Nieto, Bryan Landgraf, Josep Samitier, Liesbet Van Heirstraeten, Klaus Stefan Drese, Marion Ritzi-Lehnert |
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| Přispěvatelé: | Publica |
| Rok vydání: | 2014 |
| Předmět: |
DNA
Bacterial Lysis Biomedical Engineering Analytic Sample Preparation Methods Bioengineering 02 engineering and technology Biology Bacterial Physiological Phenomena 01 natural sciences Biochemistry Microbiology chemistry.chemical_compound Automation Lab-on-a-Chip Humans Sample preparation Respiratory Tract Infections Respiratory tract infections Bacteria 010401 analytical chemistry Nucleic acid methods General Chemistry DNA Microfluidic Analytical Techniques 021001 nanoscience & nanotechnology 0104 chemical sciences Community-Acquired Infections chemistry Influenza A virus Nucleic acid extraction RNA RNA Viral Human medicine RNA extraction 0210 nano-technology |
| Zdroj: | Lab on a chip |
| ISSN: | 1473-0189 1473-0197 |
| Popis: | In this paper, we describe the development of an automated sample preparation procedure for etiological agents of community-acquired lower respiratory tract infections (CA-LRTI). The consecutive assay steps, including sample re-suspension, pre-treatment, lysis, nucleic acid purification, and concentration, were integrated into a microfluidic lab-on-a-chip (LOC) cassette that is operated hands-free by a demonstrator setup, providing fluidic and valve actuation. The performance of the assay was evaluated on viral and Gram-positive and Gram-negative bacterial broth cultures previously sampled using a nasopharyngeal swab. Sample preparation on the microfluidic cassette resulted in higher or similar concentrations of pure bacterial DNA or viral RNA compared to manual benchtop experiments. The miniaturization and integration of the complete sample preparation procedure, to extract purified nucleic acids from real samples of CA-LRTI pathogens to, and above, lab quality and efficiency, represent important steps towards its application in a point-of-care test (POCT) for rapid diagnosis of CA-LRTI. |
| Databáze: | OpenAIRE |
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