Abstract
Quantitative polymerase chain reaction (qPCR) is a well-recognized technique for amplifying and quantifying nuclear acid, and its real-time monitoring capability, ultrahigh sensitivity, and accuracy make it a "golden-standard" tool in both molecular biology research and clinical diagnostics. However, current qPCR tests rely on bulky instrumentation and skilled laboratorians in centralized laboratories, which spatially and temporally separate the sample collection and test, leading to longer sample turnaround times (TATs) and limited working conditions. Herein, we propose an integrated optical fiber real-time polymerase chain reaction (iF-PCR) system that successfully allows convenient sample collection, rapid thermocycling, closed-loop thermal annealing, and real-time fluorescence detection in a tiny capillary reactor. By leveraging the easy-handling capillary-fiber structure and rapid photothermal actuation of the graphene-decorated fiber head, the whole TAT can be shortened, including sampling and 40 thermocycle amplifications, within 23min, in which an ultrasmall sample volume of ∼15μL is needed. Furthermore, the thermal amplification and fluorescence detection capability of the optical fiber system was cross-checked by a commercial qPCR instrument. The fiber-optic qPCR strategy can correctly distinguish between positive and negative samples of clinical respiratory syncytial virus (RSV) without the need for laboratory professional skills. Taking advantage of the distance signal transmission nature of optical fibers, the proposed strategy enables remote testing, which can eliminate the necessity of instrument deployment in crowded biosafety laboratories and facilitate potential distributed pathogen testing for coping with a new round of pandemics.
Published Version
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