Detection of Odontoglossum ringspot virus Based on Measurements of Nano Particles’ Brownian Velocity
Date Issued
2011
Date
2011
Author(s)
Teng, Chiao-Ti
Abstract
The thesis reports on an approach for detecting Odontoglossum ringspot virus (ORSV) by using micro-particle-tracking-velocimetry (micro-PTV). The ORSV is one of the major viruses in orchids, for example, Phalaenopsis, which is the most important economic plant and export good in Taiwan. To sensing the ORSV, the anti-ORSV proteins are modified on the 300 nm and 500 nm fluorescent beads and then separately mixed with the coat proteins of the ORSV and ORSV samples. The nanobeads suspended in samples were traced and statistically analyzed their Brownian velocity in real-time by micro-PTV. Through observing the variation of Brownian velocity of the nanobeads, the number of anti-ORSV and ORSV conjugation can be quantitatively estimated. Moreover, the image of the ORSV and coat proteins conjugating with nanobeads were captured through transmission electron microscope (TEM) to demonstrate the approach. In the experiment, the detection limit is exhibited to the ratio nanobeads to coat proteins of 1 to 1.5 in 8 minute. Based on our current Brownian motion measurement technique using micro-PTV, the detection limit of the concentration of 0.1 μg/ml can be found. The novel method provides higher sensitivity than enzyme-linked immunosorbent assay (ELISA) and easier operating than reverse transcription polymerase chain reaction (RT-PCR) technique, which are currently major orchid virus detection methods. Further, because of the potential that less device fabrication request, it shows higher capability on integrating with any microfluidic device for making the portable device with disposable chips.
Subjects
Brownian motion
bio-sensor technique
Transmission Electron Microscope (TEM)
Micro-Particle-Tracking-Velocimetry(μ-PTV)
Odontoglossum ringspot virus
Type
thesis
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