1. Modular development of a prototype point of care molecular diagnostic platform for sexually transmitted infections
- Author
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Branavan, Manoharanehru, Mackay, Ruth E., Craw, Pascal, Naveenathayalan, Angel, Ahern, Jeremy C., Sivanesan, Tulasi, Hudson, Chris, Stead, Thomas, Kremer, Jessica, Garg, Neha, Baker, Mark, Sadiq, Syed T., and Balachandran, Wamadeva
- Subjects
Point-of-care ,Microfluidics ,Biophysics ,Biomedical Engineering ,Molecular diagnostics ,Microengineering - 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 1.32×106 of sample DNA through thermophillic helicase dependant amplification and 1×105 copy numbers Chlamydia trachomatis genomic DNA within 10min through recombinase polymerase nucleic acid amplification tests. This work is funded under the UKCRC Translational Infection Research (TIR) Initiative supported by the Medical Research Council (Grant number G0901608) with contributions to the grant from the Biotechnology and Biological Sciences Research Council, the National Institute for Health Research on behalf of the Department of Health, the Chief Scientist Office of the Scottish Government Health Directorates and the Wellcome Trust. Axxin are thanked for sup- plying the T16-ISO platform.
- Published
- 2015