title: Detection of viruses directly from the fresh leaves of a Phalaenopsis orchid using a microfluidic system.
abstract: 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.
  
  
  BACKGROUND
  In 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.
authors.affiliation: Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan. Electronic address: wenhsinchang0805@gmail.com.
authors.forename: Wen-Hsin
authors.initials: WH
authors.lastname: Chang
authors.forename: Sung-Yi
authors.initials: SY
authors.lastname: Yang
authors.forename: Chih-Lin
authors.initials: CL
authors.lastname: Lin
authors.forename: Chih-Hung
authors.initials: CH
authors.lastname: Wang
authors.forename: Ping-Chen
authors.initials: PC
authors.lastname: Li
authors.forename: Tzong-Yueh
authors.initials: TY
authors.lastname: Chen
authors.forename: Fuh-Jyh
authors.initials: FJ
authors.lastname: Jan
authors.forename: Gwo-Bin
authors.initials: GB
authors.lastname: Lee
mesh_terms: D004867:Equipment Design; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D029595:Orchidaceae / Q000821:virology*; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000821:virology*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D012367:RNA, Viral
keywords: CNC; CaCV; Capsicum chlorosis virus; CymMV; Cymbidium mosaic virus; DNA; EMVs; LOC; Loop-mediated-isothermal-amplification; MEMS; Microfluidics; ORSV; Odontoglossum ringspot virus; PCR; PDMS; PMMA; Pathogen detection; Phalaenopsis orchid; RNA; RT-LAMP; TSWV; Tomato spotted wild virus; base-pair; bp; computer-numerical-control; deoxyribonucleic acid; electromagnetic valves; lab-on-a-chip; micro-electro-mechanical-systems; polydimethylsiloxane; polymerase chain reaction; polymethylmethacrylate; reverse-transcription loop-mediated-isothermal-amplification; ribonucleic acid