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

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, concentr...

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Bibliographic Details
Published in:Biosensors & bioelectronics 2018-06, Vol.109, p.156-163
Main Authors: Kadimisetty, Karteek, Song, Jinzhao, Doto, Aoife M., Hwang, Young, Peng, Jing, Mauk, Michael G., Bushman, Frederic D., Gross, Robert, Jarvis, Joseph N., Liu, Changchun
Format: Article
Language:English
Subjects:
3D Printing
Biosensing Techniques
Cerebrospinal Fluid - microbiology
Colorimetry
Fluorescence and colorimetry
Humans
Isothermal amplification
Limit of Detection
Malaria, Falciparum - blood
Malaria, Falciparum - parasitology
Meningococcal Infections - cerebrospinal fluid
Meningococcal Infections - microbiology
Meningococcal Infections - pathology
Microfluidic chip
Microfluidics
Neisseria meningitidis - isolation & purification
Neisseria meningitidis - pathogenicity
Nucleic acid isolation
Nucleic Acids - chemistry
Nucleic Acids - isolation & purification
Pathology, Molecular
Plasmodium falciparum - isolation & purification
Plasmodium falciparum - pathogenicity
Point of care diagnostics
Point-of-Care Systems
Printing, Three-Dimensional
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Summary: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. [Display omitted] •First application of 3D printed microfluidic reactor array for molecular diagnostics.•A simple leak-proof polymerization bonding method to integrate nucleic acid isolation membranes.•Static coating was adopted to improve the biocompatibility of the 3D printed devices.•Both end-point colorimetric detection and real-time fluorescence detection.•Detection limits of 100 fg for P. falciparum in plasma & 50 CFU for N. meningitidis in CSF.
ISSN:0956-5663
1873-4235
DOI:10.1016/j.bios.2018.03.009