Journal Article

Paper microfluidic implementation of loop mediated isothermal amplification for early diagnosis of hepatitis C virus

Bradley-Stewart, Amanda, Garrett, Alice, Sabir, Suleman R., Witkowska McConnell, Weronika, Thomson, Emma C., Jajesniak, Pawel, Davis, Chris, Xu, Gaolian, Cooper, Jonathan M., Reboud, Julien, Gunson, Rory, Yang, Zhugen

2021

Overview

The early diagnosis of active hepatitis C virus (HCV) infection remains a significant barrier to the treatment of the disease and to preventing the associated significant morbidity and mortality seen, worldwide. Current testing is delayed due to the high cost, long turnaround times and high expertise needed in centralised diagnostic laboratories. Here we demonstrate a user-friendly, low-cost pan-genotypic assay, based upon reverse transcriptase loop mediated isothermal amplification (RT-LAMP). We developed a prototype device for point-of-care use, comprising a LAMP amplification chamber and lateral flow nucleic acid detection strips, giving a visually-read, user-friendly result in <40 min. The developed assay fulfils the current guidelines recommended by World Health Organisation and is manufactured at minimal cost using simple, portable equipment. Further development of the diagnostic test will facilitate linkage between disease diagnosis and treatment, greatly improving patient care pathways and reducing loss to follow-up, so assisting in the global elimination strategy. Current HCV nucleic acid-based diagnosis is largely performed in centralised laboratories. Here, the authors present a pan-genotypic RNA assay, based on reverse transcriptase loop mediated isothermal amplification and develop a low-cost prototype paper-based lateral flow device for point-of-care use, providing a visually read result within 40 min.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 45/62 / 45/71 / 45/77 / 45/88 / 45/90 / 631/326/2521 / 631/326/596/1905 / 639/166/985 / 692/700/139/1420/1340 / Assaying / Biomedical Engineering - methods / Blood Urea Nitrogen / Diagnosis / Diagnostic systems / Diagnostic Tests, Routine / Early Diagnosis / Equipment costs / Genotype / Hepacivirus / Hepatitis / Hepatitis C / Hepatitis C - diagnosis / Humanities and Social Sciences / Humans / Laboratories / Low cost / Microfluidics / Microfluidics - methods / Molecular Diagnostic Techniques - methods / Morbidity / multidisciplinary / Nucleic Acid Amplification Techniques - methods / Nucleic acids / Point-of-Care Systems / Portable equipment / Prototypes / RNA-directed DNA polymerase / Science / Science (multidisciplinary) / Viral Load / World Health Organization