Molecular diagnostics that involve nucleic acid amplification tests (NAATs) are crucial for prevention and treatment of infectious diseases. In this study, we developed a simple, inexpensive, disposable, fully 3D printed microfluidic reactor array that is capable of carrying out extraction, concentration and isothermal amplification of nucleic acids in variety of body fluids. The method allows rapid molecular diagnostic tests for infectious diseases at point of care. A simple leak-proof polymerization strategy was developed to integrate flow-through nucleic acid isolation membranes into microfluidic devices, yielding a multifunctional diagnostic platform. Static coating technology was adopted to improve the biocompatibility of our 3D printed device. We demonstrated the suitability of our device for both end-point colorimetric qualitative detection and real-time fluorescence quantitative detection. We applied our diagnostic device to detection of Plasmodium falciparum in plasma samples and Neisseria meningitides in cerebrospinal fluid (CSF) samples by loop-mediated, isothermal amplification (LAMP) within 50 min. The detection limits were 100 fg for P. falciparum and 50 colony-forming unit (CFU) for N. meningitidis per reaction, which are comparable to that of benchtop instruments. This rapid and inexpensive 3D printed device has great potential for point-of-care molecular diagnosis of infectious disease in resource-limited settings.

涉及核酸扩增测试（NAAT）的分子诊断对于预防和治疗传染病至关重要。在这项研究中，我们开发了一种简单，廉价，一次性，全3D打印的微流体反应器阵列，该阵列能够在各种体液中进行核酸的提取，浓缩和等温扩增。该方法可以在护理点对感染性疾病进行快速的分子诊断测试。开发了一种简单的防漏聚合策略，以将流通式核酸分离膜整合到微流体设备中，从而产生多功能的诊断平台。采用静态涂层技术来改善3D打印设备的生物相容性。我们证明了该设备适用于端点比色定性检测和实时荧光定量检测。我们将诊断设备应用于检测通过回路介导的等温扩增（LAMP）在50分钟内完成血浆样本中的恶性疟原虫和脑脊液（CSF）样本中的脑膜炎奈瑟球菌。每个反应的恶性疟原虫的检出限为100 fg，脑膜炎奈瑟氏菌的检出限为50个菌落形成单位（CFU），与台式仪器相当。这种快速且便宜的3D打印设备在资源有限的环境中具有传染病分子即时诊断的巨大潜力。