Modular development of a prototype point of care molecular diagnostic platform for sexually transmitted infections. Issue 8 (August 2016)

• Record Type: Journal Article
• Authors: 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.; Balachandran, Wamadeva
• Is Part Of: Medical engineering & physics. Volume 38:Issue 8(2016:Aug.)
• Page Start: 741
• Page End: 748
• Publication Date: 2016-08
• Subjects: Microfluidics -- Microengineering -- Point-of-care -- Molecular diagnostics; Biomedical engineering -- Periodicals; Biomedical Engineering -- Periodicals; Physics -- Periodicals; Génie biomédical -- Périodiques; Biomedical engineering; Electronic journals; Periodicals; 610.28
• DOI: 10.1016/j.medengphy.2016.04.022
• Languages: English
• ISSNs: 1350-4533

Abstract

Highlights: A low cost, optical, POC molecular diagnostic platform. Sample preparation using a paper membrane. Isothermal amplification using HDA and RPA. 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 0.1 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 1.32×10 6 of sample DNA through thermophillic helicase dependant amplification and 1×10 5 copy numbers Chlamydia trachomatis genomic DNA within 10 min through recombinase polymerase nucleic acid amplification tests.