title: Modular development of a prototype point of care molecular diagnostic platform for sexually transmitted infections.
abstract: This paper presents the design of a modular point of care test platform that integrates a proprietary sample collection device directly with a microfluidic cartridge. Cell lysis, within the cartridge, is conducted using a chemical method and nucleic acid purification is done on an activated cellulose membrane. The microfluidic device incorporates passive mixing of the lysis-binding buffers and sample using a serpentine channel. Results have shown extraction efficiencies for this new membrane of 69% and 57% compared to the commercial Qiagen extraction method of 85% and 59.4% for 0.1ng/µL and 100ng/µL salmon sperm DNA respectively spiked in phosphate buffered solution. Extraction experiments using the serpentine passive mixer cartridges incorporating lysis and nucleic acid purification showed extraction efficiency around 80% of the commercial Qiagen kit. Isothermal amplification was conducted using thermophillic helicase dependant amplification and recombinase polymerase amplification. A low cost benchtop real-time isothermal amplification platform has been developed capable of running six amplifications simultaneously. Results show that the platform is capable of detecting 1.32×10(6) of sample DNA through thermophillic helicase dependant amplification and 1×10(5) copy numbers Chlamydia trachomatis genomic DNA within 10min through recombinase polymerase nucleic acid amplification tests.
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Manoharanehru
authors.initials: M
authors.lastname: Branavan
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK. Electronic address: eesrrem@brunel.ac.uk.
authors.forename: Ruth E
authors.initials: RE
authors.lastname: Mackay
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK; Oceans and Atmosphere Flagship, Commonwealth Science and Industrial Research Organisation (CSIRO), Hobart, Tasmania 7001, Australia.
authors.forename: Pascal
authors.initials: P
authors.lastname: Craw
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Angel
authors.initials: A
authors.lastname: Naveenathayalan
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Jeremy C
authors.initials: JC
authors.lastname: Ahern
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Tulasi
authors.initials: T
authors.lastname: Sivanesan
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Chris
authors.initials: C
authors.lastname: Hudson
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Thomas
authors.initials: T
authors.lastname: Stead
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Jessica
authors.initials: J
authors.lastname: Kremer
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Neha
authors.initials: N
authors.lastname: Garg
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Mark
authors.initials: M
authors.lastname: Baker
authors.affiliation: Institute for Infection and Immunity, St George's University of London, Cranmer Terrace, London SW17 0RE, UK.
authors.forename: Syed T
authors.initials: ST
authors.lastname: Sadiq
authors.affiliation: College of Engineering, Design and Physical Sciences, Brunel University London, Kingston Lane, Uxbridge, UK.
authors.forename: Wamadeva
authors.initials: W
authors.lastname: Balachandran
mesh_terms: D000818:Animals; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D006358:Hot Temperature; D056656:Lab-On-A-Chip Devices; D025202:Molecular Diagnostic Techniques / Q000295:instrumentation*; D021141:Nucleic Acid Amplification Techniques; D019095:Point-of-Care Systems; D012749:Sexually Transmitted Diseases / Q000175:diagnosis*
chemical_list: D004247:DNA
keywords: Microengineering; Microfluidics; Molecular diagnostics; Point-of-care