Instrument-free point-of-care molecular diagnosis of H1N1 based on microfluidic convective PCR

Abstract

A simple, convenient, easy-to-use, and low-cost platform for rapid molecular detection of H1N1 virus is designed and implemented based on convective polymerase chain reaction (CPCR) and dipstick assay at the point of care (POC) settings. A fully disposable chemically-heated thermal processor is developed to heat the capillary tube from the bottom at a fixed temperature for amplification without the need for electrical power. Amplification is achieved when reagents in the capillary tube are spontaneously and repeatedly transported through different temperature zones, which significantly shortens the amplification time by eliminating the need for controlled time-domain temperature cycling. A rapid one-step nucleic acid dipstick assay is adopted to facilitate the detection of amplification products with the eye instead of instrumentation. The utility of our POC diagnostic system was demonstrated by detecting H1N1 virus with sensitivity of 1.0 TCID50/mL within 35min. The developed system is particularly desired for resource poor settings, for example, developing countries, outside centralized laboratories, doctor’s offices and homes, to relieve the requirements for laboratory facilities and trained personnel.