Detection of viruses directly from the fresh leaves of a Phalaenopsis orchid using a microfluidic system

Early detection of pathogens is crucial for the effective surveillance of diseases. Many efforts have been made to explore methods which can detect these pathogens within a short period of time without requiring a tedious protocol. However, these developed methods have disadvantages such as they are relatively time-consuming or require specialized laboratory facilities. In this work, we have developed an integrated microfluidic system for rapid and automatic detection of viruses by direct analysis from fresh Phalaenopsis orchid leaves. The entire protocol, including ribonucleic acid (RNA) purification, reverse transcription loop-mediated-isothermal-amplification (RT-LAMP) and optical detection by measuring changes in turbidity was performed on a single chip. This is the first time that an integrated microfluidic system for the detection of viruses infecting the Phalaenopsis orchid has been demonstrated. The sensitivity of the developed system was also explored in this study to validate its performance.From the Clinical EditorIn this study, the authors report the development of an integrated microfluidic system for rapid and automatic detection of viruses by direct analysis of fresh Phalaenopsis orchid leaves, performing the 3-step protocol using a single chip. Similar methods may find clinical application for fast and accurate detection of viral infections.

Graphical AbstractAn integrated microfluidic system for the detection of viruses infecting the Phalaenopsis orchid has been developed. The microfluidic chip was composed of three layers, air chamber layer, liquid channel layer and glass substrate. The entire protocol, including ribonucleic acid (RNA) purification, reverse transcription loop-mediated-isothermal-amplification (RT-LAMP) and optical detection by measuring changes in turbidity was performed on a single chip. This microfluidic LAMP system has a detection limit of 35 pg. This is the first time an integrated microfluidic system was reported to directly measure pathogens common to the Phalaenopsis orchid from fresh leaves.Figure optionsDownload high-quality image (230 K)Download as PowerPoint slide