Evaluating commercial thermoplastic materials in fused deposition modeling 3D printing for their compatibility with DNA storage and analysis by quantitative polymerase chain reaction
Evaluating commercial thermoplastic materials in fused deposition modeling 3D printing for their compatibility with DNA storage and analysis by quantitative polymerase chain reaction
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DOI：
10.1039/d2ay00772j
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发表时间：
2022-06-20
影响因子：
3.1
通讯作者：
Anderson，Jared L.
中科院分区：
文献类型：
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作者：
Eitzmann，Derek R.;Anderson，Jared L.
核酸在生物样品中普遍存在，并且可以使用核酸扩增测定法灵敏地检测。为了获得高度准确和可靠的结果，通常需要核酸分离和纯化，并且可能限制这些测定的可及性。将这些工作流封装到单个设备上可以通过以商用三维（3D）打印机为特征的制造方法来实现。本研究的目的是表征熔融沉积成型（FDM）丝的基础上，其与核酸储存的兼容性，使用定量聚合酶链反应（qPCR）。为了研究核酸的吸附，使用六种常见的热塑性塑料制造储存容器，包括：聚乳酸（PLA）、尼龙、丙烯腈丁二烯苯乙烯（ABS）、共聚酯（CPE）、聚碳酸酯（PC）和聚丙烯（PP）。短的98个碱基对和较长的830个碱基对片段的DNA吸附到容器的壁中显示出在聚合物材料之间以及相同聚合物的颜色品种之间显著变化。在2.5 M NaCl TE缓冲液中储存12小时后，发现PLA储存容器吸附最少量的98碱基对DNA，而ABS和PC容器吸附高达97.2 ± 0.2%和97.5 ± 0.2%。DNA吸附可以通过降低3D打印物体的层高来减少，从而增加ABS储存容器的功能。发现尼龙将qPCR抑制组分解吸到储存溶液中，导致qPCR分析的DNA定量数据错误。
Nucleic acids are ubiquitous in biological samples and can be sensitively detected using nucleic acid amplification assays. To achieve highly accurate and reliable results, nucleic acid isolation and purification is often required and can limit the accessibility of these assays. Encapsulation of these workflows onto a single device may be achieved through fabrication methodologies featuring commercial three-dimensional (3D) printers. This study aims to characterize fused deposition modeling (FDM) filaments based on their compatibility with nucleic acid storage using quantitative polymerase chain reaction (qPCR). To study the adsorption of nucleic acids, storage vessels were fabricated using six common thermoplastics including: polylactic acid (PLA), nylon, acrylonitrile butadiene styrene (ABS), co-polyester (CPE), polycarbonate (PC), and polypropylene (PP). DNA adsorption of a short 98 base pair and a longer 830 base pair fragment to the walls of the vessel was shown to vary significantly among the polymer materials as well as the color varieties of the same polymer. PLA storage vessels were found to adsorb the least amount of the 98 base pair DNA after 12 hours of storage in 2.5 M NaCl TE buffer whereas the ABS and PC vessels adsorbed up to 97.2 ± 0.2% and 97.5 ± 0.2%. DNA adsorption could be reduced by decreasing the layer height of the 3D printed object, thereby increasing the functionality of the ABS storage vessel. Nylon was found to desorb qPCR inhibiting components into the stored solution which led to erroneous DNA quantification data from qPCR analysis.