| Autor: |
Markus Jeziorski, Hans Attig, Gerd Grosshauser, Andy Wende, Klaus Stefan Drese, Thomas Rothmann, Marion Ritzi-Lehnert, Eva Schaeffer, Ralf Himmelreich, Jan Claussen, Isabell Wick, Rainer Dahlke, Ole Wiborg |
| Rok vydání: |
2010 |
| Předmět: |
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| Zdroj: |
ASME 2010 8th International Conference on Nanochannels, Microchannels, and Minichannels: Parts A and B. |
| DOI: |
10.1115/fedsm-icnmm2010-31048 |
| Popis: |
Early diagnosis followed by personalised efficient therapy of infectious diseases (e.g. respiratory diseases, meningitis, sepsis) can lead to considerable reduction of costs in health care. Point-of-care testing (POCT) can provide early detection since this kind of decentralised analysis can be done by unskilled personnel at any time. Other advantages of automated miniaturised Lab-on-a-Chip systems (LoC) are reduction of time and reagents, elimination of cross-contamination and enhanced reproducibility due to enhanced process control. Such Lab-on-a-Chip systems will establish themselves on market only when sensitivity and specificity meet clinical requirements. An integrated cost-efficient lab-on-a-chip system is presented which allows performing all diagnostic process steps for pathogen analysis of respiratory viruses from nasopharyngeal samples. The microfluidic disposable chip comprises structures for lysis of nasopharyngeal swab samples, preparation of total nucleic acids using magnetic silica beads, reverse transcription followed by QIAplex PCR technology and labelling of the nucleic acids by hybridisation with LiquiChip Beads and streptavidin-R-phycoerythrin. Labelled target sequences are transferred for analysis into a QIAGEN LiquiChip 200 workstation. The core of the instrument is a construction based on rotating heating bars allowing for fast cycling. All chemicals needed for performing of 24 analyses are either stored freeze-dried on the single-use disposable microfluidic chip (processing cartridge) or as liquids in a separate reagent cartridge. After introducing the sample into the lysis chamber of the microfluidic chip and inserting the chip into the device all steps are done automatically. To realise these steps, fluidic control in terms of light barriers and turning valves are integrated into the injection moulded disposable chip. This includes metering structures as well as magnetic stir bars for mixing. The functionality was proven by direct comparison of samples processed manually vs. automatically using the “ResPlex Panel II” for detection of respiratory viruses from nasopharyngeal samples. The efficiency of the automated LoC system yields at about 30–60% as compared to the manually performed reference experiments. Comparing the performance of the instrument with commercially available kits and nucleic acid preparation devices showed slightly weaker but clearly positive final signal intensities obtained from the prototype device even without protocol optimization.Copyright © 2010 by ASME |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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