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Title (Primary) An integrated lab-on-a-chip for preconcentration of bacteria and nucleic acid extraction
Author Hügle, M.; Dame, G.; Behrmann, O.; Rietzel, R.; Karthe, D.; Hufert, F.T.; Urban, G.A.;
Journal RSC Advances
Year 2018
Department ASAM;
Volume 36
Issue 8
Language englisch;
POF III (all) T32;
Abstract To improve detection sensitivity, molecular diagnostics require preconcentration of low concentrated samples followed by rapid nucleic acid extraction. This is usually achieved by multiple centrifugation, lysis and purification steps, for instance, using chemical reagents, spin columns or magnetic beads. These require extensive infrastructure as well as time consuming manual handling steps and are thus not suitable for point of care testing (POCT). To overcome these challenges, we developed a microfluidic chip combining free-flow electrophoretic (FFE) preconcentration (1 ml down to 5 μl) and thermoelectric lysis of bacteria as well as purification of nucleic acids by gel-electrophoresis. The integration of these techniques in a single chip is unique and enables fast, easy and space-saving sample pretreatment without the need for laboratory facilities, making it ideal for the integration into small POCT devices. A preconcentration efficiency of nearly 100% and a lysis/gel-electrophoresis efficiency of about 65% were achieved for the detection of E. coli. The genetic material was analyzed by RT-qPCR targeting the superfolder Green Fluorescent Protein (sfGFP) transcripts to quantify mRNA recovery and qPCR to determine DNA background.
ID 20440
Persistent UFZ Identifier http://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=20440
Hügle, M., Dame, G., Behrmann, O., Rietzel, R., Karthe, D., Hufert, F.T., Urban, G.A. (2018):
An integrated lab-on-a-chip for preconcentration of bacteria and nucleic acid extraction
RSC Advances 36 (8), 20124 - 20130