Rotary-based platform with disposable fluidic modules for automated isolation of nucleic acids

We describe the development and evaluation of a rotary-based platform with multiple disposable fluidic modules for simultaneous automatic nucleic acid (NA) isolation from up to 24 biological samples. The procedure is performed inside insulated individual disposable modules, which minimizes both the risk of infection of personnel and laboratory cross-contamination. Each module is a segment of a circular cylinder containing a leak-proof inlet port for sample input, reservoirs with lyophilized chemicals and solvents, fluidic channels, stoppers, valves, a waste reservoir and an outlet port equipped with the standard micro test tube for NA collection. The entire platform, apart from the rotor that accommodates 24 modules, consists of functional elements that provide spinning of the rotor, reagent mixing, pressure delivery, and heating of reaction mixtures. The transfer of the reaction mixtures inside the modules is performed either with rotation of the rotor or with excessive air pressure applied to the module’s reservoirs. The entire process takes less than 40 min, starting from the sample loading to the recovery of the purified NA, and it allows NA isolation both from bacterial cells and viral particles. The feasibility and reproducibility of the developed platform was demonstrated by the NA isolation from suspensions of Bacillus thuringiensis and Mycobacterium tuberculosis cells within a concentration range of 108 to 102 cells/ml. Isolation of NAs from blood plasma samples with varying concentration of hepatitis B and C viruses from 107 to 102 particles/ml were also successful. The purity and integrity of the extracted NAs were both reliable for performing quantitative PCR.

[1]  M. Ratain,et al.  A Novel Fully Automated Molecular Diagnostic System (AMDS) for Colorectal Cancer Mutation Detection , 2013, PloS one.

[2]  Tae Seok Seo,et al.  A rotary microsystem for simple, rapid and automatic RNA purification. , 2012, Lab on a chip.

[3]  Marc Madou,et al.  A novel, compact disk-like centrifugal microfluidics system for cell lysis and sample homogenization. , 2007, Colloids and surfaces. B, Biointerfaces.

[4]  R. Kaiser,et al.  Extraction of viral nucleic acids: comparison of five automated nucleic acid extraction platforms. , 2012, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[5]  Marc Madou,et al.  Lab on a CD. , 2006, Annual review of biomedical engineering.

[6]  M. Salimans,et al.  Rapid and simple method for purification of nucleic acids , 1990, Journal of clinical microbiology.

[7]  A. Makarov,et al.  Microfluidic module for automated isolation and purification of nucleic acids from biological samples , 2010, Doklady Biochemistry and Biophysics.

[8]  M. Kodani,et al.  Comparison of commercial systems for extraction of nucleic acids from DNA/RNA respiratory pathogens , 2010, Journal of Virological Methods.

[9]  Jintae Kim,et al.  Centrifugal microfluidics for biomedical applications. , 2010, Lab on a chip.

[10]  A. Kloke,et al.  The LabTube - a novel microfluidic platform for assay automation in laboratory centrifuges. , 2014, Lab on a chip.

[11]  Jack T Trevors,et al.  Perspective: microfluidic applications in microbiology. , 2010, Journal of microbiological methods.

[12]  R. Zengerle,et al.  Microfluidic lab-on-a-chip platforms: requirements, characteristics and applications. , 2010, Chemical Society reviews.

[13]  Yoon-Kyoung Cho,et al.  One-Step Pathogen Specific DNA Extraction from Whole Blood on a Centrifugal Microfluidic Device , 2007 .

[14]  A. Mirzabekov,et al.  Evaluation of hybridisation on oligonucleotide microarrays for analysis of drug-resistant Mycobacterium tuberculosis. , 2005, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[15]  S. Berensmeier Magnetic particles for the separation and purification of nucleic acids , 2006, Applied Microbiology and Biotechnology.

[16]  K. Peñuelas-Urquides,et al.  Measuring of Mycobacterium tuberculosis growth. A correlation of the optical measurements with colony forming units , 2013, Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology].

[17]  Martin A M Gijs,et al.  Microfluidic applications of magnetic particles for biological analysis and catalysis. , 2010, Chemical reviews.

[18]  Dominic Eicher,et al.  Microfluidic devices for diagnostic applications , 2011, Expert review of molecular diagnostics.

[19]  Manjima Dhar,et al.  Research highlights: microfluidic point-of-care diagnostics , 2014 .

[20]  R. Zengerle,et al.  Frequency-dependent transversal flow control in centrifugal microfluidics. , 2005, Lab on a chip.

[21]  Mauro Ferrari,et al.  Point-of-care technologies for molecular diagnostics using a drop of blood. , 2014, Trends in biotechnology.

[22]  Young Ki Choi,et al.  A microbead-incorporated centrifugal sample pretreatment microdevice. , 2013, Lab on a chip.

[23]  A. Zasedatelev,et al.  Gel-based microarrays in clinical diagnostics in Russia , 2011, Expert review of molecular diagnostics.

[24]  M. Bergeron,et al.  Validation of a centrifugal microfluidic sample lysis and homogenization platform for nucleic acid extraction with clinical samples. , 2010, Lab on a chip.

[25]  Roland Zengerle,et al.  Centrifugal LabTube platform for fully automated DNA purification and LAMP amplification based on an integrated, low-cost heating system , 2014, Biomedical microdevices.

[26]  Roland Zengerle,et al.  Centrifugal gas-phase transition magnetophoresis (GTM)--a generic method for automation of magnetic bead based assays on the centrifugal microfluidic platform and application to DNA purification. , 2013, Lab on a chip.

[27]  James P Landers,et al.  Purification of nucleic acids in microfluidic devices. , 2008, Analytical chemistry.

[28]  D. Gryadunov,et al.  Method for automated extraction and purification of nucleic acids and its implementation in microfluidic system , 2011, Applied Biochemistry and Microbiology.

[29]  R. Álvarez,et al.  Comparison of the QIAsymphony automated nucleic acid extraction and PCR setup platforms with NucliSens easyMAG and Corbett CAS-1200 liquid handling station for the detection of enteric pathogens in fecal samples. , 2011, Journal of microbiological methods.