Branavan et al 2016.pdf (1.93 MB)
Modular development of a prototype point of care molecular diagnostic platform for sexually transmitted infections
journal contribution
posted on 2023-05-18, 21:28 authored by Branavan, M, Mackay, RE, Pascal Craw, Naveenathayalan, A, Ahern, JC, Sivanesan, T, Hudson, C, Stead, T, Kremer, J, Garg, N, Baker, M, Sadiq, ST, Balachandran, WThis 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 0.1 ng/µL and 100 ng/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 1.32×106 of sample DNA through thermophillic helicase dependant amplification and 1×105 copy numbers Chlamydia trachomatis genomic DNA within 10 min through recombinase polymerase nucleic acid amplification tests.
History
Publication title
Medical Engineering and PhysicsVolume
38Issue
8Pagination
741-748ISSN
1350-4533Department/School
School of Natural SciencesPublisher
Elsevier Sci LtdPlace of publication
The Boulevard, Langford Lane, Kidlington, Oxford, England, Oxon, Ox5 1GbRights statement
Copyright 2016 The Author(s). Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/Repository Status
- Open