Fully 3D printed integrated reactor array for point-of-care molecular diagnostics

Abstract

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.

An ultraportable and versatile point-of-care DNA testing platform

“…Because of our combinatory and integrated approach, the POCKET system was extremely light and small, rendering the POCKET system, to the best of our knowledge, to be one of the smallest and most portable POC DNA testing devices. Compared with other 3D-printed or microfluidic chips, our POCKET system was more versatile for a broader range of possible sample sources and achieved multiplex detections with reduced reagent costs (table S6) (6,9,11,14,17,(27)(28)(29)(30)(31).…”

3D Printed Biosensor Arrays for Medical Diagnostics

“…In previous examples, 3D printing was utilized to fabricate microfluidics that served only one Reproduced with permission from [9]. Copyright (2018) Elsevier.…”

“…3D printing impacts many design aspects including high resolution, low cost and fast fabrication of complex microfluidic devices in a continuous process [9,10,11]. 3D printed microfluidic devices have been used to fabricate semi and fully automated diagnostic approaches for diseases like cancer [12,13], infectious diseases [14,15,16], and xenobiotic genotoxicity [17].…”

“…The device was powered by micropumps that were fully automated to reduce variation and make it a good point of care diagnostic approach. Another multifunctional microfluidic device was also introduced recently by Kadimisetty et al[9], where they were able to extract, concentrate and isothermally amplify nucleic acids in different body fluids as an approach for microfluidic point of care diagnostics (Figure 8B). The microfluidic device was integrated with a membrane to isolate nucleic acids, then placed in a chamber where loop mediated isothermal amplification is induced.…”

3D-Printed Biosensor Arrays for Medical Diagnostics

“…In this review, we focus on applications of 3D printing techniques in medical diagnostics. We discuss different 3D printing techniques and how these techniques impact many design aspects including resolution, cost and fabrication of complex diagnostic devices in a continuous process [ 9 , 10 , 11 ]. 3D-printed microfluidic devices have been used to fabricate semi and fully automated diagnostic approaches for diseases like cancer [ 12 , 13 ], infectious diseases [ 14 , 15 , 16 ], and xenobiotic genotoxicity [ 17 ].…”