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High-throughput roll-to-roll production of polymer biochips for multiplexed DNA detection in point-of-care diagnostics
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-10-22 , DOI: 10.1039/d0lc00751j
Pelin Toren 1 , Martin Smolka 1 , Anja Haase 1 , Ursula Palfinger 1 , Dieter Nees 1 , Stephan Ruttloff 1 , Ladislav Kuna 1 , Cindy Schaude 1 , Sandra Jauk 1 , Markus Rumpler 2 , Bettina Hierschlager 3 , Ingo Katzmayr 3 , Max Sonnleitner 3 , Manuel W Thesen 4 , Mirko Lohse 4 , Martin Horn 5 , Wilfried Weigel 5 , Matija Strbac 6 , Goran Bijelic 6 , Suhith Hemanth 7 , Nastasia Okulova 7 , Jan Kafka 7 , Stefan Kostler 1 , Barbara Stadlober 1 , Jan Hesse 1
Affiliation  

Roll-to-roll UV nanoimprint lithography has superior advantages for high-throughput manufacturing of micro- or nano-structures on flexible polymer foils with various geometries and configurations. Our pilot line provides large-scale structure imprinting for cost-effective polymer biochips (4500 biochips/hour), enabling rapid and multiplexed detections. A complete high-volume process chain of the technology for producing structures like μ-sized, triangular optical out-couplers or capillary channels (width: from 1 μm to 2 mm, height: from 200 nm up to 100 μm) to obtain biochips (width: 25 mm, length: 75 mm, height: 100 μm to 1.5 mm) was described. The imprinting process was performed with custom-developed resins on polymer foils with resin thicknesses ranging between 125–190 μm. The produced chips were tested in a commercial point-of-care diagnostic system for multiplexed DNA analysis of methicillin resistant Staphylococcus aureus (e.g., mecA, mecC gene detections). Specific target DNA capturing was based on hybridisation between surface bound DNA probes and biotinylated targets from the sample. The immobilised biotinylated targets subsequently bind streptavidin–horseradish peroxidase conjugates, which in turn generate light upon incubation with a chemiluminescent substrate. To enhance the light out-coupling thus to improve the system performance, optical structures were integrated into the design. The limits-of-detection of mecA (25 bp) for chips with and without structures were calculated as 0.06 and 0.07 μM, respectively. Further, foil-based chips with fluidic channels were DNA functionalised in our roll-to-roll micro-array spotter following the imprinting. This straightforward approach of sequential imprinting and multiplexed DNA functionalisation on a single foil was also realised for the first time. The corresponding foil-based chips were able to detect mecA gene DNA sequences down to a 0.25 μM concentration.

中文翻译:

高通量生产聚合物生物芯片,用于即时诊断中的多重DNA检测

卷对卷UV纳米压印光刻技术具有高通量,可在具有各种几何形状和配置的柔性聚合物箔上高吞吐量地制造微结构或纳米结构。我们的中试线可为经济高效的聚合物生物芯片(每小时4500个生物芯片)提供大规模的结构印迹,从而实现快速和多重检测。完整的大批量工艺链技术,可生产诸如微米级,三角形光学输出耦合器或毛细管通道(宽度:1μm至2 mm,高度:200 nm至100μm)之类的结构,以获得生物芯片(描述了宽度:25mm,长度:75mm,高度:100μm至1.5mm。压印过程是使用定制开发的树脂在聚合物箔上进行的,树脂厚度在125-190μm之间。金黄色葡萄球菌例如mecAmecC基因检测)。特异性靶标DNA捕获基于表面结合的DNA探针与样品中生物素化靶标之间的杂交。固定的生物素化靶标随后结合抗生蛋白链菌素-辣根过氧化物酶结合物,与化学发光底物一起孵育后会发光。为了增强光输出耦合从而改善系统性能,将光学结构集成到设计中。mecA的检出限具有和不具有结构的芯片的(25 bp)分别计算为0.06和0.07μM。此外,在压印后,在我们的卷对卷微阵列点样仪中将具有流体通道的基于箔的芯片DNA功能化。这种在单箔纸上进行顺序印记和DNA多重化的直接方法也是第一次实现。相应的基于箔的芯片能够检测低至0.25μM浓度的mecA基因DNA序列。
更新日期:2020-11-03
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