Lab-on-a-Chip Multiplex Assays
Lab-on-a-chip multiplex assays allow a rapid identification of multiple parameters in an automated manner. Here we describe a lab-based preparation followed by a rapid and fully automated DNA microarray hybridization and readout in less than 10 min using
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Introduction There is an urgent need for rapid diagnostics in the field of infectious diseases where fast decisions need to be made for an effective therapy. Traditional microbiological methods take around 48 h for species identification and antibiogram results. The use of molecular tests built on multi-parameter lab-on-a-chip platforms are a good option to speed up the time taken between sampling and results. Here we describe a protocol which makes use of the Fraunhofer integrated lab-on-a-chip in vitro diagnostic (ivD) platform. The platform can be used to run multiplex immunoassays and serological assays to identify antibodies directly from blood samples [1]. In this protocol we demonstrate another type of application, consisting of a fully automated DNA microarray assay and readout, together with a lab-based preparation method. This protocol allows rapid bacterial species identification and the genotyping of relevant antibiotic resistance genes. The advantage of microarrays is the possibility to analyze a large amount of parameters at one time, making these assays particularly useful in the field of antibiotic resistance determination [2–7].
Paul C. Guest (ed.), Multiplex Biomarker Techniques: Methods and Applications, Methods in Molecular Biology, vol. 1546, DOI 10.1007/978-1-4939-6730-8_25, © Springer Science+Business Media LLC 2017
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Nonetheless, there are still some disadvantages regarding current DNA microarray protocols that hinder the broad use of this approach in routine analysis. These include long hybridization times and complex laboratory based procedures, as well as the need for skilled operational personnel. The use of lab-on-a-chip based systems may help to overcome these limitations since they offer a rapid and automated solution, which integrates many of the lab-based procedures. The combination of microarrays with microfluidics constitutes an elegant solution to automate and speed up microarray hybridization. The ivD platform consists of a microfluidic cartridge and a base unit. The credit card sized cartridge contains all relevant elements necessary for DNA microarray hybridization. These are mainly reservoirs for all of the reagents, integrated pumping systems, the specific microarray, integrated temperature control for hybridization and an optical transducer to allow integrated sensing [1, 8, 9]. The base unit contains all necessary electronics to control the cartridge, an optical readout system to analyze the microarray directly after hybridization and a touch screen to control the assay and monitor the results. With this setup, hybridization times of less than 5 min can be achieved, yielding equivalent results as found by lab-based microarray hybridizations of 1 h. Therefore, the total time to generation of results is less than 10 min for hybridization, washing and readout. Here, we describe all necessary steps from sample preparation to microarray analysis for a lab-based and lab-on-a-chip-based procedure. As an example, we use a species identification m
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