title: Activated paper surfaces for the rapid hybridization of DNA through capillary transport.
abstract: The development of low-cost, accurate, and equipment-free diagnostic tests is crucial to many clinical, laboratory, and field applications, including forensics and medical diagnostics. Cellulose fiber-based paper is an inexpensive, biodegradable, and renewable resource, the use of which as a biomolecule detection matrix and support confers several advantages compared to traditional materials such as glass. In this context, a new, facile method for the preparation of surface functionalized papers bearing single-stranded probe DNA (ssDNA) for rapid target hybridization via capillary transport is presented. Optimized reaction conditions were developed that allowed the direct, one-step activation of standard laboratory filters by the inexpensive and readily available bifunctional linking reagent, 1,4-phenylenediisothiocyanate. Such papers were thus amenable to subsequent coupling of amine-labeled ssDNA under standard conditions widely used for glass-based supports. The intrinsic wicking ability of the paper matrix facilitated rapid sample elution through arrays of probe DNA, leading to significant, detectable hybridization in the time required for the sample liquid to transit the vertical length of the strip (less than 2 min). The broad applicability of these paper test strips as rapid and specific diagnostics in "real-life" situations was exemplified by the discrimination of amplicons generated from canine and human mitochondrial and genomic DNA in mock forensic samples.
authors.affiliation: Division of Glycoscience, School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Center, SE-106 91 Stockholm, Sweden.
authors.forename: Ana Catarina
authors.initials: AC
authors.lastname: Araújo
authors.forename: Yajing
authors.initials: Y
authors.lastname: Song
authors.forename: Joakim
authors.initials: J
authors.lastname: Lundeberg
authors.forename: Patrik L
authors.initials: PL
authors.lastname: Ståhl
authors.forename: Harry
authors.initials: H
authors.lastname: Brumer
mesh_terms: D000588:Amines / Q000737:chemistry; D000818:Animals; D002482:Cellulose / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000235:genetics*; D004285:Dogs; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010209:Paper; D013499:Surface Properties; D013997:Time Factors
chemical_list: D000588:Amines; D015342:DNA Probes; D004277:DNA, Single-Stranded; D002482:Cellulose