title: Polyethersulfone improves isothermal nucleic acid amplification compared to current paper-based diagnostics.
abstract: Devices based on rapid, paper-based, isothermal nucleic acid amplification techniques have recently emerged with the potential to fill a growing need for highly sensitive point-of-care diagnostics throughout the world. As this field develops, such devices will require optimized materials that promote amplification and sample preparation. Herein, we systematically investigated isothermal nucleic acid amplification in materials currently used in rapid diagnostics (cellulose paper, glass fiber, and nitrocellulose) and two additional porous membranes with upstream sample preparation capabilities (polyethersulfone and polycarbonate). We compared amplification efficiency from four separate DNA and RNA targets (Bordetella pertussis, Chlamydia trachomatis, Neisseria gonorrhoeae, and Influenza A H1N1) within these materials using two different isothermal amplification schemes, helicase dependent amplification (tHDA) and loop-mediated isothermal amplification (LAMP), and traditional PCR. We found that the current paper-based diagnostic membranes inhibited nucleic acid amplification when compared to membrane-free controls; however, polyethersulfone allowed for efficient amplification in both LAMP and tHDA reactions. Further, observing the performance of traditional PCR amplification within these membranes was not predicative of their effects on in situ LAMP and tHDA. Polyethersulfone is a new material for paper-based nucleic acid amplification, yet provides an optimal support for rapid molecular diagnostics for point-of-care applications.
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.|Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, 47907, USA.
authors.forename: J C
authors.identifier: http://orcid.org/0000-0003-4962-0908
authors.initials: JC
authors.lastname: Linnes
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.
authors.forename: N M
authors.initials: NM
authors.lastname: Rodriguez
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.
authors.forename: L
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA. catherin@bu.edu.
authors.forename: C M
authors.initials: CM
authors.lastname: Klapperich
mesh_terms: D004265:DNA Helicases / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D019937:Diagnostic Techniques and Procedures; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics; D008567:Membranes, Artificial; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010209:Paper; D019095:Point-of-Care Systems; D011108:Polymers / Q000737:chemistry*; D016062:Porosity; D012367:RNA, Viral / Q000235:genetics; D013450:Sulfones / Q000737:chemistry*
chemical_list: D004269:DNA, Bacterial; D008567:Membranes, Artificial; D011108:Polymers; D012367:RNA, Viral; D013450:Sulfones; C022840:polyether sulfone; D004265:DNA Helicases
keywords: Isothermal nucleic acid amplification; Paper-based diagnostic device; Point-of-care diagnostics; Polyethersulfone