Labor im Zentrifugenröhrchen zur dezentralen patientennahen Diagnostik
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1 Labor im Zentrifugenröhrchen zur dezentralen patientennahen Diagnostik Felix von Stetten HSG-IMIT, Georges Köhler Alle 103, Freiburg, Germany Mobile Diagnostik am Point-of-Care, DGBMT, Düsseldorf
2 HSG-IMIT Institut für Mikro- und Informationstechnik Freiburg & Villingen-Schwenningen, Germany 120 Employees (FTE) 13 Mio turnover Quality Management DIN EN IS0 9001:2008 Business units Lab-on-a-Chip Microfluidics Sensors & Systems Prototyping & Production Freiburg Villingen-Schwenningen Felix von Stetten Mobile Diagnostik am DGBMT 2
3 Reasons for Point of Care Testing < 2 days Turnaround time Centralized laboratory Opportunities POC test Rapid decision making goal directed therapy Reduced cost for treatment, care & hospitalization Reduced morbidity, mortality & spread of antibiotic resistant bacteria Felix von Stetten Mobile Diagnostik am DGBMT 3
4 Point of Care Platforms Requirements Easy to use at the point of care Microfluidic cartridge Integration into diagnostic process chain Results comparable to those of centralized lab Components Microfluidic unit-use cartridge Assay processing Pre-stored reagents Processing device e.g. LabDisk Platform (HSG-IMIT, Qiagen Lake Constance) Processing device extremely specialized, expensive development & production Felix von Stetten Mobile Diagnostik am DGBMT 4
5 Technological approach to cost reduction POCT that can be processed by standard laboratory devices Sample Diagnostik-Kit Centrifuge tube Mixing chamber Extraction matrix Product Reagent prestorage revolver Processing revolver, rotated by pen-mechanics Collection revolver Dry reagent Detector Laboratory centrifuge Waste Felix von Stetten Mobile Diagnostik am DGBMT 5
6 Processing of LabTube cartridge Spring Force Centrifuge, spinning protocol controlled via RS232 Interface Centrifugal Force LabTube ballpen mechanism Felix von Stetten Mobile Diagnostik am DGBMT 6
7 Processing steps Example: DNA purification Sample in L P B - W1 W2 - E Mixing chamber Stepwise actuation by ball pen mechanism 8x Silica column Waste Eluate Felix von Stetten Mobile Diagnostik am DGBMT 7
8 Processing steps Supply of lysis buffer L P B - W1 W2 - E etc. 8x 8x Mixing chamber Silica column Waste Eluate Felix von Stetten Mobile Diagnostik am DGBMT 8
9 Proof of principle: Liquid handling Reagent revolver Storage L S B W 1 W 2 E Lanced sealing foil Collection revolver 3 cm Storage Lysis-buffer Sample Binding Wash 1 Wash 2 Elution Felix von Stetten Mobile Diagnostik am DGBMT 9
10 Proof of principle: DNA extraction Sample: Lysed rape seed Hands on time: 6 min 1 min Felix von Stetten Mobile Diagnostik am DGBMT 10
11 Proof of principle: Protein purification Sample: lysed insect cells; Protein: Histidyl trna synthetase Reference kd LabTube L W 1 W 2 W 3 E 1 E 2 M M L W 1 W 2 W 3 E 1 E 2 Felix von Stetten Mobile Diagnostik am DGBMT 11
12 In progress BMBF-Project Easy Tube Immunoassay based sepsis test Quantitation of sepsis biomarkers / PCT, Parallel processing of 6-8 channels 2 ml whole blood sample Photometric detection Anticipated time to result minutes Decision about medication within 1h Felix von Stetten Mobile Diagnostik am DGBMT 12
13 In progress BMBF-Project Easy Tube Sample to answer nucleic acid based MRSA sepsis testing Pathogen isolation and identification Lysis of blood cells (degradation of background DNA) Pathogen enrichment Pathogen DNA extraction DNA amplification Anticipated time to result 50 min Decision about antibiotic therapy within 1h 2 h Felix von Stetten Mobile Diagnostik am DGBMT 13
14 In progress BMBF-Project Easy Tube Sample to answer nucleic acid based MRSA sepsis testing Anticipated workflow Sample Functional packaging of reagents Laboratory centrifuge Amplification / Detection Felix von Stetten Mobile Diagnostik am DGBMT 14
15 Cost per extraction Cost evaluation of lab tube based analysis (e.g. DNA extraction) Rough evaluation includes Instrument (5 year invest) Hands-on-time Reagents Disposables LabTube is beneficial for Medium level sample throughput Number of samples per year Manual (incl. centrifuge invest) Automated via QIAcube Automated via LabTube Felix von Stetten Mobile Diagnostik am DGBMT 15
16 Summary LabTube: point of care testing based on standard laboratory centrifuge Integrated reagent prestorage Fully automated assay processing Interfaces to diagnostic process chain Rapid and less expensive development Nucleic acid extraction and protein purification demonstrated Hands on time decreased by the factor of 6 MRSA sepsis test under development Felix von Stetten Mobile Diagnostik am DGBMT 16
17 Acknowledgement Easy-Tube consortium BMBF-Projekt: 16SV5451K Laufzeit: 06/ /2014 MicroLab consortium MicroLab, FKZ HSG/50 Felix von Stetten Mobile Diagnostik am DGBMT 17
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