Branavan, M;
Mackay, RE;
Craw, P;
Naveenathayalan, A;
Ahern, JC;
Sivanesan, T;
Hudson, C;
Stead, T;
Kremer, J;
Garg, N;
et al.
Branavan, M; Mackay, RE; Craw, P; Naveenathayalan, A; Ahern, JC; Sivanesan, T; Hudson, C; Stead, T; Kremer, J; Garg, N; Baker, M; Sadiq, ST; Balachandran, W
(2016)
Modular development of a prototype point of care molecular diagnostic platform for sexually transmitted infections.
Medical Engineering & Physics, 38 (8).
pp. 741-748.
ISSN 1873-4030
https://doi.org/10.1016/j.medengphy.2016.04.022
SGUL Authors: Sadiq, Syed Tariq
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.
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