School of Engineering, University of Leicester, Leicester, LE1 7RH, UK; Department of Biomedical Engineering, College of Engineering, Al-Nahrain University, Baghdad, Iraq.
State Key Laboratory of Advanced Design and Manufacturing for Vehicle Body, Hunan University, 410082, China.
Microbiol Res. 2020 Dec;241:126587. doi: 10.1016/j.micres.2020.126587. Epub 2020 Aug 21.
Novel sampling matrices were manufactured using 3D printing for the detection of respiratory pathogens in expired air. A specific configuration of the matrices was designed using Computer-Aided Design software. Polyvinyl alcohol (PVA) was printed using fused deposition modelling to create a multilayer matrix to enhance the capture of bacteria. The performance of these matrices was compared with gelatine filters that have been used for this work to date. PVA matrices (60 mm diameter) were contaminated with bacteria either by direct inoculation, or by aerosol exposure using an Omron A3 nebuliser. Rough and smooth morphotypes of Mycobacterium abscessus, M. smegmatis and M. bovis BCG, were used in this study to contaminate the matrices. PVA matrices and gelatine sampling filters were contaminated to compare recovery rates for quantitative analyses. These were dissolved in water, bacteria pelleted and DNA extracted followed by a Mycobacterium-specific quantitative Polymerase Chain Reaction (qPCR).The results showed that 3D printed PVA matrices are very effective to capture the bacteria. 3D printed PVA matrix and gelatine filters yielded results of the same order of magnitude for mycobacterial analyses, however, PVA matrix offers several advantages over the latter material. 3D printed PVA is considered as an economic and time-effective matrix as it is cheaper than gelatine filters. PVA is sufficiently robust to be handled and loaded into the surgical masks for sampling, compared to the brittle gelatine filters that required supportive frames. PVA is a synthetic material and it is suitable for DNA-based analyses, whilst gelatine is derived from animal collagen, and carries a high bacterial DNA background that interferes with the target DNA analysis. Furthermore, PVA dissolves in distilled water without requiring chemicals or enzymes, such as the case for gelatine hydrolysis. To summarise, 3D printed PVA sampling matrix is considered a promising tool used for microbiological diagnostic purposes.
新型采样矩阵使用 3D 打印技术制造，用于检测呼出空气中的呼吸道病原体。使用计算机辅助设计软件设计了矩阵的特定配置。使用熔融沉积建模技术打印聚乙烯醇（PVA），以创建多层矩阵来增强细菌的捕获。将这些矩阵的性能与迄今为止用于此工作的明胶过滤器进行了比较。PVA 矩阵（直径 60 毫米）通过直接接种或使用欧姆龙 A3 雾化器进行气溶胶暴露来污染细菌。在这项研究中，使用粗糙和光滑形态的脓肿分枝杆菌、耻垢分枝杆菌和牛分枝杆菌卡介苗来污染矩阵。PVA 矩阵和明胶采样过滤器被污染以比较定量分析的回收率。将它们溶解在水中，沉淀细菌并提取 DNA，然后进行分枝杆菌特异性定量聚合酶链反应（qPCR）。结果表明，3D 打印的 PVA 矩阵非常有效地捕获细菌。3D 打印的 PVA 矩阵和明胶采样过滤器对分枝杆菌分析的结果具有相同的数量级，但 PVA 矩阵相对于后者具有几个优势。3D 打印的 PVA 被认为是一种经济有效的矩阵，因为它比明胶过滤器便宜。与易碎的明胶过滤器相比，PVA 足够坚固，可以进行处理和装入外科口罩进行采样。PVA 是一种合成材料，适用于基于 DNA 的分析，而明胶源自动物胶原蛋白，并且具有较高的细菌 DNA 背景，会干扰目标 DNA 分析。此外，PVA 可在不使用化学物质或酶（如明胶水解）的情况下溶解于蒸馏水中。总之，3D 打印的 PVA 采样矩阵被认为是一种有前途的用于微生物诊断目的的工具。
